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Fluorescence microscopy is a ubiquitous technique in the life sciences that uses fluorescent molecules to visualize specific components of biological specimens. This powerful tool has revolutionized biology, and it represents a perfect example of the advancements enabled by biophysical research and technology development. However, despite its central role in contemporary research, fluorescence is hardly covered in typical secondary school curricula, with few hands-on “entry-level” materials available for secondary school teachers to introduce this important method to their students. Furthermore, most commercially available fluorescence microscopes are prohibitively costly and often appear as “black boxes.” To address this gap, we introduce here an experimental, research-grade fluorescence microscopy kit and educational resource targeted at secondary school students and teachers. Microscope in Action is an interdisciplinary resource based on active learning that combines concepts from both optics and biology. The students assemble a functional microscope from basic optical, mechanical, and electronic parts, thereby testing and understanding the function of each component “hands-on.” We also present sample preparation and imaging activities that can be incorporated to enable an exploration of biological topics with the assembled microscope and exercises in which students actively learn and practice scientific thinking by collecting and analyzing data. Although the resource was developed with secondary schools in mind, the variety of available protocols and the adjustable module lengths make it suitable for different age groups and topics, from middle school to PhD level, from short workshops to courses spanning several days.

Background Cardiac computed tomography (CT) emerged as an accurate tool for non-invasive evaluation of coronary artery disease (CAD), being able to identify high risk non-calcified atherosclerosis. Identification of high risk CAD in its asymptomatic stage could be an interest target for medical therapy. Nowadays no validated tools are available to predict the presence of high risk atherosclerosis, probably due to the multifactorial pathogenesis of atherosclerosis. Facial features may express both genetic and environmental factors that could be associated to high risk atherosclerosis. Aim of the present study was to verify whether deep learning models applied to facial features may accurately predict the presence of high risk coronary atherosclerosis evaluated at cardiac CT Methods We enrolled a consecutive cohort of patients who underwent clinical indicated cardiac CT for suspected, CAD. Before CT, 10 facial photos were taken from every patients from random fronts views. All cardiac CT were analysed for the presence of non-calcified plaque volume (defined as 23 mm3 was used to define a patients with an elevated volume non-calcified plaque We built a deep learning model, exploiting the transfer learning technique; briefly, we implemented an “xception” architecture, joining a pre-trained convolutional part with a specific combination of dense layers, in which an output layer follows a hidden layer with 512 neurons and a dropout layer with a dropout rate=0.2. The batch size, the number of epochs and the learning rate were 16, 20, and 0.0001, respectively. A training set composed of 198 face images was fed into the model, while 20 face images served as test set for the prediction of the presence of elevated volume of non-calcified plaque from patients facial features. Results We present early results from the first 20 patients enrolled (12 male and 8 female, with mean age of 73±13 years old). In 9 patients cardiac CT resulted completely normal, while in 11 subjects the presence of coronary atherosclerosis was demonstrated. Among them, 9 patients presented non-calcified coronary atherosclerosis, while 6 had an elevated volume of non-calcified plaque. On the test set, we obtained an accuracy, sensitivity, specificity, positive predictive value, negative predictive values and and AUC equal to 0.90, 1, 0.8, 0.83, 1, and 0.99, respectively for the prediction of the presence of an elevated volume of non-calcified plaque from facial features among all 20 patients enrolled. Conclusions Prediction of the presence of high risk atherosclerosis from deep learning models applied to facial features appeared to be feasible and promising. Our results may provide a useful tool for appropriate identification of patients that may merit to underwent cardiac CT, even if asymptomatic, for early identification of high risk atherosclerosis Funding Acknowledgement Type of funding sources: None.

Publisher Copyright: © This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited In this contribution, new data concerning the distribution of vascular flora alien to Italy are presented. It includes new records, confirmations, exclusions, and status changes for Italy or for Italian administrative regions. Nomenclatural and distribution updates published elsewhere are provided as Suppl. material 1.

In this contribution, new data concerning the distribution of vascular flora alien to Italy are presented. It includes new records, confirmations, exclusions, and status changes for Italy or for Italian administrative regions. Furthermore, three new combinations are proposed. Nomenclatural and distribution updates published elsewhere are provided as Suppl. material 1.

Background In patients with systolic chronic heart failure (CHF) clinical signs of congestion cannot always be evident at clinical examination. Right atrial pressure (RAP) measured by right heart catheterization (RHC) is an accurate and reproducible marker of blood volume. A non-invasive accurate tool to identify CHF patients with normal RAP would be desirable to tailor therapy. Purpose To validate an ultrasound (US)-assessed internal jugular vein distensibility (JVD) ratio to identify patients with normal mean RAP (defined as 7 mmHg or less) measured by RHC. Methods We first identify the JVD ratio that allows the most accurate identification of patients with normal RAP in a prospective calibration cohort of 100 patients with systolic CHF. Then, we tested the JVD ratio threshold to identify patients with normal RAP in a validation cohort of 101 consecutive patients with systolic CHF. All patients had a left ventricular ejection fraction (LVEF) Results In the calibration cohort (mean age 53 years, 13% female; median LVEF 25%, 81% in NYHA class III/IV) we define the best threshold of the JVD ratio to identify patients with normal RAP that has 1.6 with an area under the curve (AUC of 0.74; p1.6, n=42; median RAP 4 mmHg). High JVD ratio and low JVD ratio groups had similar clinical and laboratory characteristics. In the validation cohort (mean age 55 years, 13% female; median LVEF 25%; 56% in NYHA class III/IV) using the previously defined 1.6 JVD ratio threshold, we identified 51 patients with low JVD ratio (median RAP 8 mmHg) and 50 patients with high JVD ratio (median RAP 3 mmHg; p Conclusions We found that US-assessed JVD ratio is a convenient and accurate diagnostic tool to identify patients with advanced systolic CHF with normal vs. increased RAP. This tool could be tested in the ambulatory setting to modulate therapies, particularly diuretics and vasodilators. Figure 1 Funding Acknowledgement Type of funding source: None

In this contribution, new data concerning the distribution of native vascular flora in Italy are presented. It includes new records, confirmations, exclusions, and status changes to the Italian administrative regions. Two new combinations are proposed. Nomenclatural and distribution updates, published elsewhere, and corrigenda are provided as Suppl. material 1.

Grapevine yellows (GYs) is one of the most damaging phytoplasma-associated diseases that causes severe yield losses in every geographic area where grapevines are cultivated. The main yellows diseases in grapevine in Europe are “flavescence dorée” (FD, 16SrV-C/D ribosomal subgroups) and “bois noir” (BN, 'Candidatus Phytoplasma solani', 16SrXII-A ribosomal subgroup), transmitted by Scaphoideus titanus Ball and Hyalestes obsoletus Signoret, respectively. Recently the mosaic leafhopper Orientus ishidae (Matsumura) (Cicadellidae; Deltocephalinae) was found to be positive to 16SrV-C and -D phytoplasmas in Slovenia, Italy and Switzerland ; it was also shown as capable to transmit 16SrV phytoplasmas from broadbean to grapevine. To verify the reasons of the continuous GY spreading in the Veneto region (North-Eastern Italy), “Prosecco areas”, the identification and molecular characterization of phytoplasmas in symptomatic and asymptomatic grapevine and insects captured in selected vineyards during a three year-survey was carried out

Introduction The Insomnia Severity Index is a commonly used instrument to assess the presence of insomnia symptoms as well as an outcome measure following an intervention. Longitudinal measurement invariance is a necessary property of an assessment instrument when it is repeated over time. The validity of conclusions regarding change in the construct ‘insomnia severity’ depend on scale equivalence at each measurement timepoint. Assessment of measurement invariance of the ISI in sleep apnea patients has never been performed. Methods Veterans with sleep apnea (n=654; AHI=36±28; 93% male; age=52±12; BMI=33±6) completed the ISI on the night of their overnight PSG and again when they picked up their PAP device. Invariance was determined by imposing a series of more restrictive equivalence constraints on a 2-factor model of the ISI. The series of constraints tested for configural, weak, strong and strict invariance. Invariance testing was modeled with exploratory structural equation modeling in Mplus (v. 7.0). Results The 2-factor model that emerged from the analysis showed items relating to nighttime symptoms loading on factor 1 and daytime symptoms loading on factor 2. The sleep ‘satisfaction’ item, however, had weak but similar loadings on both factors. The increasingly restrictive constraints imposed on the model revealed no decrement in model fit (RMSEA=.039 to.043; CFI=.987 to .980; TLI=.981-.977; SRMR=.027-.041). Conclusion The ISI met strict criteria for longitudinal measurement invariance demonstrating that it is a valid instrument to be used in repeated measures study designs of insomnia in sleep apnea patients. Change over time on the ISI is not due to the changing measurement characteristics of the ISI but to true changes in the ‘insomnia severity’ construct. Support None

An updated inventory of the vascular flora alien to Italy, providing details on the occurrence at regional level, is presented. The checklist includes 1597 species, subspecies, and hybrids, distributed in 725 genera and 152 families; 2 taxa are lycophytes, 11 ferns and fern allies, 33 gymnosperms, and 1551 angiosperms. 157 taxa are archaeophytes and 1440 neophytes. The alien taxa currently established in Italy are 791 (570 naturalized and 221 invasive), while 705 taxa are casual aliens, 4 are not assessed, 7 are of unknown regional distribution, 47 have not been confirmed in recent times, 3 are considered extinct or possibly extinct in the country, and 40 are doubtfully occurring in Italy. This checklist allows to establish an up-to-date number (9792) of taxa constituting the whole (native and alien) Italian flora.

An updated inventory of the vascular flora alien to Italy, providing details on the occurrence at regional level, is presented. The checklist includes 1597 species, subspecies, and hybrids, distributed in 725 genera and 152 families; 2 taxa are lycophytes, 11 ferns and fern allies, 33 gymnosperms, and 1551 angiosperms. 157 taxa are archaeophytes and 1440 neophytes. The alien taxa currently established in Italy are 791 (570 naturalized and 221 invasive), while 705 taxa are casual aliens, 4 are not assessed, 7 are of unknown regional distribution, 47 have not been confirmed in recent times, 3 are considered extinct or possibly extinct in the country, and 40 are doubtfully occurring in Italy. This checklist allows to establish an up-to-date number (9792) of taxa constituting the whole (native and alien) Italian flora.

In this contribution new data concerning the distribution of native vascular flora in Italy are presented. It includes new records, exclusions, and confirmations to the Italian administrative regions for taxa in the generaAsplenium,Bolboschoenus,Botrychium,Chamaerops,Crocus,Galeopsis,Grafia,Helosciadium,Hieracium,Juniperus,Leucanthemum,Lolium,Medicago,Phalaris,Piptatherum,Potamogeton,Salicornia,Salvia,Seseli,Silene,Spiraea,TorilisandVicia.Rhaponticoidescalabricais proposed assynonym novumofR.centaurium. Furthermore, new combinations in the generaGalatellaandLactucaare proposed.

Excerpt from the Proceedings of the Tenth Annual Acquisition Research Symposium Acquisition Management

This paper will examine a sandstone cliff south of the town of Livorno Italy, between the Rio Maroccone mouth to the north and the Leone cove to the south, in its most dangerous area, between the sea and the main road Aurelia. The aim of this paper is to divide the area into homogenous hazard classes and assess the possible kinematic evolution. The paper refers to an evaluation of parameters that may trigger rock falls on the cliff, which could constitute dangers for tourists. The identification of critical areas has been achieved through the elaboration of landslide susceptibility maps. The geotechnical study was carried out on the Macigno Fm at the site. Field surveys and indirect tests for classifying rock masses and estimating the uniaxial compression strength (UCS) were performed. Some numerical analyses were executed to study the static and dynamic behavior of the slope. The results confirm the vulnerability of the areas, demonstrating the dynamic instability of the analyzed sections and a precarious equilibrium is also in static conditions. These analyses, compared with the site surveys, show poor data and in some cases alarming results; some portions of the cliff are under limit equilibrium conditions or close to it, especially in some of the areas appreciated by tourists and bather. Through the numerical modeling the critical situation of these slopes becomes even more evident, confirmed by some rock falls of small blocks occurred during the geomechanical survey period. To make the situation even worse, the whole area is constantly subject to exogenous agents such as strong winds and the wave action, which determine a rapid deterioration of the mechanical characteristics of strength in the Macigno Sandstone.

Biogeographia vol. XX V//I - 2007 {Pubblicato il 30 dicembre 2007) Biogeografia de/l’Appennino centrale e settentriona/e: trenfanni dopo Fitogeografla apuana. Aggiornamenti biosistematici, tassonomici e corologici sulle Spermatophyta endemiche, relitte e critiche FABIO GARBARI, GIANNI BEDINI, l\/IARIAANSALDI, DINO l\/lARCHETTl* Orto laotzmico eMu5eo /aotzmim, Dz'p:zrn'mm2f0 di Biologizz, z/izz L. G/:zz'm' 5, 56126 Pisa (Italy); e—mzzi[: fgm‘[7zzrz'@[7i0[ogz'zz. zzizzpi. it, gb£tz'z'nz'@[2z'0[0gz'zz. zmzpi. it, mzzn.tzz/a'z'@/7i0/0gz'zz. Zt7ll;Z7li. it *Vizz Irorzzo 6, 54100 Mzzsszz (Itzz/y); e—mzzz'[: dz'rz0mzzrcbettz'@/i[m*0.it Key words: Apuan Alps, biosystematics. endemics, relics, plant distribution SUMMARY An updated review of the biosystematics, chorology and taxonomy of the main important taxa of the Apuan Alps flowering plants is given, particularly of the endemic, relic and rare units. Of some of them a distribution map is provided, together with information on their population demography, ecological preferences and karyology. INTRODUZIONE A piii di 35- anni di distanza dal convegno della Societet Italiana di Biogeografia dedicato alle Alpi Apuane, E: stato rivisto il quadro delle coho- scenze floristiche di questo settore dell’Italia appenninica, i cui tratti salienti erano stati delineati a quel tempo da Ferrarini (1971), da Orsino (1971), da Pichi Serrnolli (1971) e, per quanto riguarda qualche aspetto citotasson0rni- co, da Garbari (1971). Nei tre decenni e mezzo trascorsi, sono state fatte numerose scoperte con descrizione di nuove unitit sistematiche o revisioni di carattere tassor1omico— nomenclaturale che hanno portato a consistenti chiarimenti del quadro floristi— co, ma anche osservazioni di tipo storico, erbariale e di carnpagna, con Valuta- zioni di carattere ecologico, fitogeografico e biosistematico qui riassunte. Le ricer- che ~ particolarmente dei botanici pisani e fiorentini — hanno portato a un buon

Between 1999 and 2001, 355 hospital laboratories in Italy were asked to complete a questionnaire addressing mycobacterial test methods, 1-year workloads and laboratory safety features. Analysis of the data showed that rapid methods for mycobacterial testing were being used by most larger laboratories; however, sub-optimal methods were still in use in small and medium-size laboratories. In a country such as Italy, which has a low prevalence of tuberculosis cases, implementation of rapid technologies, combined with regionalisation of mycobacterial diagnostic services, seems to be the most reasonable and costeffective strategy.

Neurotrophins (NTs) modulate the brain invasion of melanoma cells and the activity of an extracellular matrix degradative enzyme, heparanase, that has been recently cloned. Heparanase degrades the heparan sulfate proteoglycans (HSPGs) and is a critical mediator of tumor metastasis and angiogenesis. Because astrocytes are among the first brain cells encountered by extravasating melanoma cells, they may play important roles in the development of brain metastases. To test this hypothesis, we used purified in vitro astrocyte cultures and found that they express heparanase transcript and functional enzyme that were up-regulated by the prototypic NT, nerve growth factor. Coincubation of astrocytes (or their conditioned medium) with brain-metastatic cells resulted in a superadditive effect on heparanase activity and up to an 8-fold increase of in vitro chemoinvasion using purified HSPGs. These observations indicate that astrocytes significantly contribute to the brain colonization of melanoma cells via heparanase-driven modalities.

Heparanase is an endo-beta-D-glucuronidase, the enzymatic targets of which are the glycosaminoglycan chains of heparan sulfate proteoglycans. Elevated levels of heparanase are associated with the metastatic potential of melanoma cells. Treatment of murine and human melanoma cells with the prototypic neurotrophin nerve growth factor (NGF) increases the production of heparanase by melanoma cells. We reported previously that physiological concentrations of NGF increased in vitro Matrigel invasion of early-passage human brain-metastatic 70W melanoma cells but not melanoma cells metastatic to other sites or nonmetastatic melanoma cells. Here we found that treatment of 70W melanoma cells with neurotrophin NT-3 increased Matrigel invasion, whereas treatment with neurotrophins other than NGF or NT-3 did not influence invasion. Mutants of NGF that do not bind to the neurotrophin receptor p75NTR or other nonneuronal growth factors were not able to enhance the invasion of 70W melanoma cells. When 70W cells were exposed to antisense oligonucleotides directed against p75NTR mRNA, there was a reduction in NGF and NT-3 binding, and the neurotrophins failed to enhance Matrigel invasion. To study the properties of heparanase in NT-regulated malignant melanoma invasive processes, we developed a sensitive heparanase assay consisting of purified [35S]heparan sulfate subpopulations separated by agarose gel electrophoresis. Incubation of 70W cells with NGF or NT-3, but not brain-derived NT factor, NT-4/5, or mutant NGF, resulted in increased release of heparanase activity that was capable of degrading a subpopulation of heparan sulfate molecules.

Abnormal expression of proteoglycans has been implicated in cancer and metastasis primarily because these macromolecules are involved in the control of cell growth and matrix assembly. In this report, we have investigated the expression and immunolocalization of perlecan, a major heparan sulfate proteoglycan of basement membranes and pericellular matrices, in human metastatic melanomas. Twenty-six of the 27 tumor samples showed a significant increase (up to 15-fold) in the perlecan mRNA levels when compared with normal tissue. This change correlated with a vast deposition of perlecan protein core in the pericellular matrix of metastatic melanomas. Furthermore, we have established a relationship between perlecan expression in clonal melanoma cells (70W) stimulated with neurotrophins and their increased invasiveness. Interestingly, perlecan mRNA levels were up-regulated within 10 min of neurotrophin stimulation, indicating that perlecan is an early response gene. This upregulation also occurred prior to heparanase production, suggesting that perlecan expression and its regulation might play a pivotal role in the initial onset of invasion.

A collaborative study was made to determine alkaline phosphatase in casein samples by the rapid colorimetric test. Six to eight collaborators tested 10 unknown casein samples containing various amounts of residual phosphatase with and without the addition of magnesium acetate. Results indicated that magnesium acetate significantly increased phosphatase activity. The collaborators correctly analyzed 95% of the samples with the added magnesium acetate. The method has been adopted official first action.

DINO MARCHETTI Istituto Botanico dell’Universit:2t - Via P.A. Mattioli, 4 » 53100 Siena Note floristiche tosco-liguri-emiliane. I. Piante indigene nuove 0 rare per le coste spezzine Con questo articolo inizia una serie di contribuni sulla flora indigena ed esotica di Liguria, Emilia e Toscana, anche se 1’atten- zione rnaggiore sare Volta soprattutto alla provincia della Spezia e alla Toseana nord-occidentale. Verranno prese in considerazione 1e segnalazioni floristiche di un certo interesse e sare fatto il riesame critioo delle entite che nelle popolazioni locali mostrano un disaccordo pifi o meno netto con le descrizioni ufficiali. In quest’ambit~o, e prevista la descrizione di qualche entite attualmente allo studio, Che mostra di possedere una sicura individualite a livello tassonomico. La prima nota riguarda investigazioni condotte principalmente alle Cinque Terre e zone contigue, per 1’esattezza lungo la costa e negli irnmediati pendii retrostanti, fine in cresta, dalla Punta del Mesco a Portovenere, con marginali sc-onfinamenti ed eventuali cenni per 1’iso1a della Palmaria. In qualche case, 0 per qualche entite, vengono presi in considerazione i dintorni orientali della Spezia, fino a1 fiume Magra e alla Punta Bianca. Tali territori sono caratterizzati da clima mite con precipitazioni atmosferiche di va- lore rnedio Che tendono a ridursi ne1l’estrernite rneridionale. Le quote sono generalrnente modeste e culminano col M. Mal-pertuso (812 m), sopra Vernazza. Attorno a Monterosso al Mare compaiono le ofioliti (serpentina e soprattutto gabbro) mentre per il resto delle Cinque Terre e fine a Campiglia -prevalgono 1e arenarie (ABBATE, 1969). Da qui a Portovenere si alternano calcari vari (ZACCAGNA, 1928). Tra La Spezia, il Magra e la Punta Bianca si hanno arenarie, quarziti e calcari (ZACCAGNA, 1928), con verrucano ne1l’estr.ema punta (TREVISAN, 1970).

Conditions were established for single cell analysis of glucocorticoid receptor (GR) content by flow cytometry using several clones of a human leukemic cell line (CCRF-CEM). These included CEM-7A, 7R, C1, and ICR 27 Tk.3 cells which were examined both by standard [3H]dexamethasone radiometric binding and by two independent flow cytometry assays. The latter involved either mouse monoclonal antibody against GR (GR-MoAb) or fluoresceinated cortisol ligand probes. For CEM-7A, 7R, and C1 cells, there was a correlation between GR-MoAb and radiometrically defined GR values. However, clone ICR-27 Tk.3 with low [3H]dexamethasone binding exhibited the highest GR-MoAb fluorescence. The fluoresceinated cortisol assay correlated with dexamethasone binding values in all four clones. Thus, GR-MoAb identifies the total immunologically reactive GR present, while the fluoresceinated cortisol assay quantifies only the functionally intact GR in terms of its initial binding. Their combined use may reveal the cellular heterogeneity of GR expression and function also in human tumor samples, to which they have been successfully applied. When coupled with DNA counterstaining, GR expression can be related directly to frequently DNA-aneuploid tumor cells and cell cycle distribution.

In Italy, although vaccination has been recommended for a number of years, vaccination coverage for mumps is still sub-optimal. The objective of the present study was to evaluate the seroprevalence of mumps antibodies in the Italian population, stratified by age, gender and geographical area. The proportion of individuals positive for mumps antibodies remained stable in the age classes 0–11 months and 1 year (25.4% and 30.8%, respectively) and showed a continuous increase after the second year of life. The percentage of susceptible individuals was higher than 20% in persons 2–14 years of age and exceeded 10% in persons 15–39 years of age. No statistically significant differences were observed by gender or geographical area. Comparison between these results and the data obtained from a 1996 survey showed a statistically significant increase in seroprevalence in the age class 2–4 years. No changes were observed in the other age-groups. The results of this study confirm that the efforts made in recent years to improve vaccination coverage within the second year of life should be strengthened.