Preliminary results in the automated detection of operational modal properties of the Portico Varano in the Camerino Ducal Palace

Portico Varano in the Ducal Palace of Camerino (Italy) is a renaissance monumental quadriporticus that was damaged by the 2016 Central Italy seismic sequence. Within the field activities for saving cultural heritage foreseen within a recent European research project named ARCH, a long-term monitoring system, comprising different types of sensors, such as accelerometers, displacement transducers, environmental sensors, and a weather station, was set up to achieve comprehensive measures of its operational behaviour and the evolution of the damage. The monitoring system, installed in October 2020, is currently operating, providing valuable information on the experimentally observed dynamic behaviour, also considering changes in the environmental conditions. Starting from the results of the dynamic characterization of the structure and after the optimization of the position of the sensors, this paper shows and discusses the efforts made to track over time the modal characteristics of the Portico Varano in order to detect changes in its conditions. In addition, a procedure has been proposed and implemented combining information available from Italian National Institute of Geophysics and Volcanology (INGV) to identify recorded data related to seismic events relevant to operational conditions

Identification and characterization of PlAlix, the Alix homologue from the Mediterranean sea urchin Paracentrotus lividus.

The sea urchin provides a relatively simple and tractable system for analyzing the early stages of embryo development. Here, we use the sea urchin species, Paracentrotus lividus, to investigate the role of Alix in key stages of embryogenesis, namely the egg fertilization and the first cleavage division. Alix is a multifunctional protein involved in different cellular processes including endocytic membrane trafficking, filamentous (F)-actin remodeling, and cytokinesis. Alix homologues have been identified in different metazoans; in these organisms, Alix is involved in oogenesis and in determination/differentiation events during embryo development. Herein, we describe the identification of the sea urchin homologue of Alix, PlAlix. The deduced amino acid sequence shows that Alix is highly conserved in sea urchins. Accordingly, we detect the PlAlix protein cross-reacting with monoclonal Alix antibodies in extracts from P. lividus, at different developmental stages. Focusing on the role of PlAlix during early embryogenesis we found that PlAlix is a maternal protein that is expressed at increasingly higher levels from fertilization to the 2-cell stage embryo. In sea urchin eggs, PlAlix localizes throughout the cytoplasm with a punctuated pattern and, soon after fertilization, accumulates in larger puncta in the cytosol, and in microvilli-like protrusions. Together our data show that PlAlix is structurally conserved from sea urchin to mammals and may open new lines of inquiry into the role of Alix during the early stages of embryo development

MITOCHONDRIAL MASS, DISTRIBUTION AND ACTIVITY DURING SEA URCHIN OOGENESIS

The sea urchin egg is a favourite model for studies of the molecular biology and physiology of fertilization and early development, yet we know sparingly little of its oocytes and of mitochondria behaviour during oogenesis. The process of oogenesis in most echinoderms is asynchronous so each ovary lobe has hundreds of oocytes at all stages of development. At the beginning of oogenesis, the oocyte is about 10 \ub5m in diameter. During the vitellogenic phase of oogenesis, the oocyte accumulate yolk proteins and grow to ten times their original size to 80 to 100 \ub5m in sea urchins. The oocyte, arrested at the prophase of the first meiotic division, is apparent with its large nucleus, the germinal vesicle (GV), containing a prominent nucleolus. Echinoid (such as sea urchin) and Holothurian oocytes complete meiotic maturation prior to fertilization, distinct from other echinoderms and almost all others animals. As maturation progresses, it occurs the GV breaks down (GVBD). These eggs may then be stored for weeks to months within the female before they are spawned, and the proportion of eggs in the ovary increases from early to late season, as the numbers of oocytes decline [1]. Mitochondria, generally known as the powerhouses of eukaryotic cells, play a primary role in cellular energetic metabolism, homeostasis and death. These organelles, with their multicopy genome maternally inherited, are directly involved at several levels in the reproductive process since their functional status influences the quality of oocytes and contributes to the process of fertilization and embryonic development. It has been demonstrated that the number of maternal mitochondria is sufficient to support development until late stages without new synthesis of mitochondrial DNA or production of new organelles [2]. During embryogenesis mitochondrial mass does not change, whereas mitochondrial respiration increases [3]. The behaviour of these organelles during oogenesis remains at moment unclear. In the present paper we studied, by Confocal Laser Scanning Microscopy tecnologies (CLSM), the mass and distribution, the activity and the DNA content of sea urchin Paracentrotus lividus mitochondria during oogenesis, by in vivo incubating oocytes of different size with cell-permeant probes specific for mitochondria and for DNA and by immunodetection of hsp60 chaperonine, a well known mitochondrial marker. In particular the oocytes were grouped in six classes: < 10, 20/30, 40/50, 60/70, 80/90 \ub5m, and 90 \ub5m ovulated egg, on the base of diameters. Microscopic observations were performed capturing 2 \ub5m thick layers of oocytes. Of the several thousands oocytes we observed, 20 for each different oogenesis stage were analyzed and processed. In order to interpret results and to draw unequivocal conclusions, we measured by IMAGE J software analysis the intensity values of fluorescent signals, as suggested in Agnello et al 2008 [4]. The mitochondria of oocytes with a diameter between 20 and 70 \ub5m, appeared to give rise to clusters that disappear in that of 80 \ub5m. In the oocytes between 60 and 90 \ub5m the red fluorescence seems to be more evident around the germinal vesicle (the merge tends to red), suggesting an increasing oxidative phosphorylation activity. In the ovulated eggs, red and green fluorescence are uniformly distributed suggesting that mitochondria are dispersed in the cytoplasm. In addition the merge of green and red colours shows that the whole mitochondrial population is consuming oxygen at the same level (the resulting colours tends to yellow), figure 1. In order to calculate the total mitochondrial mass and activity we integrated the values of pixel intensities for all captured sections and used the arithmetic means to draw a statistical analysis. Results suggest a parallel rise of mitochondrial mass and activity, suggesting that the amount and activity of organelles change remarkably during oogenesis. Figure 1. shows the distribution of hsp60 protein, detected by immunofluorescence analysis (A), the mitochondrial and genomic DNA, after in vivo incubation with PicoGreen probe (B) and the merge of green and red fluorescence signal, respectively due to mitochondrial mass and activity, after in vivo incubation with Mitotraker Green and Orange (C). The size of the oocytes reported is 80 \ub5m. Results suggest that mitochondria are actively duplicating and that mitochondrial DNA is replicating during the different oogenesis phases. It is noteworthy that around the germinal vesicle, especially in the larger oocytes, next to the germinal vesicle breakdown, the organelles are more active in oxygen consumption, probably due to the major energetic needing in this key moment of gametogenesis. [1] Wessel G.M., Voronina E., and Brooks J.M. (2004) Obtaining and handling echinoderm oocytes. In \u201cMethods in Cell Biology\u201d, Elsevier. Vol.74, Chapter 5, pp. 87-114. [2] Matsumoto L., Kasamatsu H., Pik\ub4o L. and Vinograd J. (1974) Mitochondrial DNA replication in sea urchin oocytes. J. Cell Biol. 63: 146\u2013159. [3] Morici G., Agnello M., Spagnolo F., Roccheri M.C., Di Liegro C.M. and Rinaldi A.M. (2007) Confocal microscopy study of the distribution, content and activity of mitochondria during Paracentrotus lividus development. Journal of Microscopy. 228: 165-173. [4] Agnello M., Morici G., Rinaldi A.M. (2008) A method for measuring mitochondrial mass and activity . Cytotechnology. 56: 145-149. Maria Carmela Roccheri: Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche. Universit\ue0 degli Studi di Palermo, Viale delle Scienze Ed.16, Palermo, Italy; tel: 09123897414; e-mail: [email protected]

Endurance training damages small airway epithelium in mice.

RATIONALE: In athletes, airway inflammatory cells were found to be increased in induced sputum or bronchial biopsies. Most data were obtained after exposure to cold and dry air at rest or during exercise. Whether training affects epithelial and inflammatory cells in small airways is unknown. OBJECTIVES: To test whether endurance training under standard environmental conditions causes epithelial damage and inflammation in the small airways of mice. METHODS AND MEASUREMENTS: Formalin-fixed, paraffin-embedded lung sections were obtained in sedentary (n = 14) and endurance-trained (n = 16) Swiss mice at baseline and after 15, 30, and 45 days of training. The following variables were assessed (morphometry and immunohistochemistry) in small airways (basement membrane length < 1 mm): (1) integrity, proliferation, and apoptosis of bronchiolar epithelium; and (2) infiltration, activation, and apoptosis of inflammatory cells. MAIN RESULTS: Compared with sedentary mice, bronchiolar epithelium of trained mice showed progressive loss of ciliated cells, slightly increased thickness, unchanged goblet cell number and appearance, and increased apoptosis and proliferation (proliferating cell nuclear antigen) (p < 0.001 for all variables). Leukocytes (CD45(+) cells) infiltrated airway walls (p < 0.0001) and accumulated within the lumen (p < 0.001); however, apoptosis of CD45(+) cells did not differ between trained and sedentary mice. Nuclear factor-kappaB translocation and inhibitor-alpha of NF-kappaB (IkappaBalpha) phosphorylation were not increased in trained compared with sedentary mice. CONCLUSIONS: Bronchiolar epithelium showed damage and repair associated with endurance training. Training increased inflammatory cells in small airways, but inflammatory activation was not increased. These changes may represent an adaptive response to increased ventilation during exercise

Push ‘o ver: in situ pushover tests on as built and strengthened existing brickwork constructions

In the last decades several experimental tests were performed to analyse the seismic capacity of unreinforced masonry, mostly involving small size structural elements or small-scale building models, not accounting for the actual complexity of existing constructions. This paper illustrates experimental pushover tests on two very similar portions of an existing masonry building. The first portion was tested as built, while the second one was tested after a strengthening intervention with Composite Reinforced Mortar (CRM) system with Glass Fiber Reinforced Polymer (GFRP) components. The retrofit consisted in the application of a reinforced plaster on both sides of the walls of the first storey and on the external side only of the second storey, a solution that preserves the continuity of use. Comparisons between the experimental response of the two portions as well as the observed behaviour of the intervention at the first storey (both wall sides) and second storey (only outer side) provide very interesting insights and preliminary information on the degree of upgrading, essential steps to support the assessment of seismic vulnerability reduction

Circulating haematopoietic and endothelial progenitor cells are decreased in COPD

Circulating CD34+ cells are haemopoietic progenitors that may play a role in tissue repair. No data are available on circulating progenitors in chronic obstructive pulmonary disease (COPD). Circulating CD34+ cells were studied in 18 patients with moderate-to-severe COPD (age: mean+/-sd 68+/-8 yrs; forced expiratory volume in one second: 48+/-12% predicted) and 12 controls, at rest and after endurance exercise. Plasma concentrations of haematopoietic growth factors (FMS-like tyrosine kinase 3 (Flt3) ligand, kit ligand), markers of hypoxia (vascular endothelial growth factor (VEGF)) and stimulators of angiogenesis (VEGF, hepatocyte growth factor (HGF)) and markers of systemic inflammation (tumour necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-8) were measured. Compared with the controls, the COPD patients showed a three-fold reduction in CD34+ cell counts (3.3+/-2.5 versus 10.3+/-4.2 cells.microL-1), and a 50% decrease in AC133+ cells. In the COPD patients, progenitor-derived haemopoietic and endothelial cell colonies were reduced by 30-50%. However, four COPD patients showed progenitor counts in the normal range associated with lower TNF-alpha levels. In the entire sample, CD34+ cell counts correlated with exercise capacity and severity of airflow obstruction. After endurance exercise, progenitor counts were unchanged, while plasma Flt3 ligand and VEGF only increased in the COPD patients. Plasma HGF levels were higher in the COPD patients compared with the controls and correlated inversely with the number of progenitor-derived colonies. In conclusion, circulating CD34+ cells and endothelial progenitors were decreased in chronic obstructive pulmonary disease patients and could be correlated with disease severity

Nuevas citas botánicas para la provincia de La Pampa (Argentina)

Blackstonia perfoliata, Huds. (Gentianaceae) y Samolus valerandi L. (Primulaceae), new records for La Pampa province are reported with information about life forms, habitat and common names.Se cita por primera vez Blackslonia perfoliata, Huds. (Gentianaceae) y Samolus valerandi L. (Primulaceae) para la provincia de La Pampa. Se consignan sus formas de vida, ambientes y nombres vulgares

Exercise intolerance at high altitude (5050 m): critical power and W'

The relationship between work rate (WR) and its tolerable duration (t(LIM)) has not been investigated at high altitude (HA). At HA (5050 m) and at sea level (SL), six subjects therefore performed symptom-limited cycle-ergometry: an incremental test (IET) and three constant-WR tests (% of IET WR(max), HA and SL respectively: WR(1) 70±8%, 74±7%; WR(2) 86±14%, 88±10%; WR(3) 105±13%, 104±9%). The power asymptote (CP) and curvature constant (W') of the hyperbolic WR-t(LIM) relationship were reduced at HA compared to SL (CP: 81±21 vs. 123±38 W; W': 7.2±2.9 vs. 13.1±4.3 kJ). HA breathing reserve (estimated maximum voluntary ventilation minus end-exercise ventilation) was also compromised (WR(1): 25±25 vs. 50±18 l min(-1); WR(2): 4±23 vs. 38±23 l min(-1); WR(3): -3±18 vs. 32±24 l min(-1)) with near-maximal dyspnea levels (Borg) (WR(1): 7.2±1.2 vs. 4.8±1.3; WR(2): 8.8±0.8 vs. 5.3±1.2; WR(3): 9.3±1.0 vs. 5.3±1.5). The CP reduction is consistent with a reduced O(2) availability; that of W' with reduced muscle-venous O(2) storage, exacerbated by ventilatory limitation and dyspnea. Copyright © 2011 Elsevier B.V. All rights reserved

Time from adenosine di-phosphate receptor antagonist discontinuation to coronary bypass surgery in patients with acute coronary syndrome: meta-analysis and meta-regression

BACKGROUND: Adenosine di-phosphate receptor antagonists (ADPRAs) blunt hemostasis for several days after administration. This effect, aimed at preventing cardiac ischemic complications particularly in patients with acute coronary syndromes (ACS), may increase perioperative bleeding in the case of cardiac surgery. Practice Guidelines recommend withholding ADPRAs for at least 5days prior to surgery, though with a weak base of evidence. The purpose of this study was to systematically review observational and experimental studies of early or late preoperative discontinuation of ADPRAs prior to coronary artery bypass grafting (CABG) for patients with ACS. METHODS: MEDLINE, EMBASE, the Cochrane Library databases up to December 2011; and reference lists. Observational and experimental studies that compared early ADPRA discontinuation with late discontinuation, or no discontinuation, in patients with ACS undergoing CABG. RESULTS: There were 19 studies, including 14,046 participants, 395 deaths and 309 reoperations due to bleeding. ADPRA late discontinuation up to CABG was associated with an increased risk of postoperative mortality (OR 1.46, 95% confidence interval (CI) 1.10 to 1.93) and reoperations due to bleeding (OR 2.18; 95% CI 1.47 to 2.62). Between-study heterogeneity was low. Meta-analysis limited to high quality or prospective studies gave consistent results. In most instances, the 95% prediction intervals for summary risk estimates confirmed the risk across study groups. CONCLUSIONS: ADPRA late discontinuation prior to CABG is associated with an increased risk of death and reoperations due to bleeding in patients with ACS. The confidence in the estimates of risk for late discontinuation is moderate to high

Fifteen years trends of cardiogenic shock and mortality in patients with diabetes and acute coronary syndromes

PURPOSE: Our study was intended to examine time trends of management and mortality of acute coronary syndrome patients with associated diabetes mellitus. METHODS: We analyzed data from 5 nationwide registries established between 2001 and 2014, including consecutive acute coronary syndrome patients admitted to the Italian Intensive Cardiac Care Units. RESULTS: Of 28,225 participants, 8521 (30.2%) had diabetes: as compared with patients without diabetes, they were older and had significantly higher rates of prior myocardial infarction and comorbidities (all P &lt; .0001). Prevalence of diabetes and comorbidities increased over time (P for trend &lt; .0001). Cardiogenic shock rates were higher in patients with diabetes, as compared with those without diabetes (7.8% vs 2.8%, P &lt; .0001), and decreased significantly over time only in patients without diabetes (P = .007). Revascularization rates increased over time in patients both with and without diabetes (both P for trend &lt; .0001), although with persistingly lower rates in patients with diabetes. All-cause in-hospital mortality was higher in patients with diabetes (5.4 vs 2.5%, respectively, P &lt; .0001) and decreased more consistently in patients without diabetes (P for trend = .007 and &lt; .0001, respectively). At multivariable analysis, diabetes remains an independent predictor of both cardiogenic shock (odds ratio 2.03; 95% confidence interval, 1.77-2.32; P &lt; .0001) and mortality (odds ratio 1.95; 95% confidence interval, 1.69-2.26; P &lt; .0001). CONCLUSIONS: Despite significant mortality reductions observed over 15 years in acute coronary syndromes, patients with diabetes continue to show threefold higher rates of cardiogenic shock and lower revascularization rates as compared with patients without diabetes. These findings may explain the persistingly higher mortality of patients with diabetes and acute coronary syndromes