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Objective: Intraoperative specimen mammography (ISM) is a diffuse technique that allows surgeons to check specimens immediately after lumpectomy. Although the specimen is slightly compressed, the radiological image can be distorted by tissue overlap, and this may affect the evaluation of tumour borders, resulting in extension of the lumpectomy. As ISM may be less precise due to inadequate compression, a vacuum effect was applied to the specimen to increase the precision of margin detection., Study Design: This study was conducted at St. Anna Hospital Breast Unit, Turin, Italy. Women who underwent lumpectomy for cancer were eligible for inclusion. Both standard ISM (sISM) and vacuum ISM (vISM) were performed. Eighteen specimens obtained after lumpectomy from 1 April 2018 to 31 April 2018 were scanned. sISM (two orthogonal projections) was performed. Next, the specimen was placed in a vacuum, and vISM was performed. The examination was completed with a second orthogonal projection after removal of the vacuum, replacement of the specimen and repositioning of the vacuum. Additional tissue was removed if the surgeon considered that excision was inadequate. Finally, the specimen was sent for definitive histopathological analysis, which is the gold standard for the assessment of surgical margins. Intraoperative histological margin assessment was not performed. The sISM and vISM images and final histopathology reports were compared., Results: For sISM, specificity was 47 % [95 % confidence interval (CI) 25-70], sensitivity was 67 % (95 % CI 21-94), positive predictive value (PPV) was 20 % (95 % CI 6-51) and negative predictive value (NPV) was 88 % (95 % CI 53-98). For vISM, specificity was 100 % (95 % CI 80-100), sensitivity was 67 % (95 % CI 21-94), PPV was 100 % (95 % CI 34-100) and NPV was 94 % (95 % CI 72-99)., Conclusion: These data suggest that the vacuum technique is feasible, cost-saving and yields results that are similar to those from frozen sections but without the limitations, such as prolonged operating time, high variability in sensitivity due to pathologists' abilities, risk of compromising the histological report, and unreliability for small lumps and ductal carcinoma in situ., Competing Interests: Declaration of Competing Interest None declared., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: Brazil ranks second worldwide in total number of COVID-19 cases and deaths. Understanding the possible socioeconomic and ethnic health inequities is particularly important given the diverse population and fragile political and economic situation. We aimed to characterise the COVID-19 pandemic in Brazil and assess variations in mortality according to region, ethnicity, comorbidities, and symptoms., Methods: We conducted a cross-sectional observational study of COVID-19 hospital mortality using data from the SIVEP-Gripe (Sistema de Informação de Vigilância Epidemiológica da Gripe) dataset to characterise the COVID-19 pandemic in Brazil. In the study, we included hospitalised patients who had a positive RT-PCR test for severe acute respiratory syndrome coronavirus 2 and who had ethnicity information in the dataset. Ethnicity of participants was classified according to the five categories used by the Brazilian Institute of Geography and Statistics: Branco (White), Preto (Black), Amarelo (East Asian), Indígeno (Indigenous), or Pardo (mixed ethnicity). We assessed regional variations in patients with COVID-19 admitted to hospital by state and by two socioeconomically grouped regions (north and central-south). We used mixed-effects Cox regression survival analysis to estimate the effects of ethnicity and comorbidity at an individual level in the context of regional variation., Findings: Of 99 557 patients in the SIVEP-Gripe dataset, we included 11 321 patients in our study. 9278 (82·0%) of these patients were from the central-south region, and 2043 (18·0%) were from the north region. Compared with White Brazilians, Pardo and Black Brazilians with COVID-19 who were admitted to hospital had significantly higher risk of mortality (hazard ratio [HR] 1·45, 95% CI 1·33-1·58 for Pardo Brazilians; 1·32, 1·15-1·52 for Black Brazilians). Pardo ethnicity was the second most important risk factor (after age) for death. Comorbidities were more common in Brazilians admitted to hospital in the north region than in the central-south, with similar proportions between the various ethnic groups. States in the north had higher HRs compared with those of the central-south, except for Rio de Janeiro, which had a much higher HR than that of the other central-south states., Interpretation: We found evidence of two distinct but associated effects: increased mortality in the north region (regional effect) and in the Pardo and Black populations (ethnicity effect). We speculate that the regional effect is driven by increasing comorbidity burden in regions with lower levels of socioeconomic development. The ethnicity effect might be related to differences in susceptibility to COVID-19 and access to health care (including intensive care) across ethnicities. Our analysis supports an urgent effort on the part of Brazilian authorities to consider how the national response to COVID-19 can better protect Pardo and Black Brazilians, as well as the population of poorer states, from their higher risk of dying of COVID-19., Funding: None., (Copyright © 2020 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 license. Published by Elsevier Ltd.. All rights reserved.)

In the healthy brain, neuronal excitability and synaptic strength are homeostatically regulated to keep neuronal network activity within physiological boundaries. Epilepsy is characterized by episodes of highly synchronized firing across in widespread neuronal populations, due to a failure in regulation of network activity. Here we consider epilepsy as a failure of homeostatic plasticity or as a maladaptive response to perturbations in the activity. How homeostatic compensation is involved in epileptogenic processes or in the chronic phase of epilepsy, is still debated. Although several theories have been proposed, there is relatively little experimental evidence to evaluate them. In this perspective, we will discuss recent results that shed light on the potential role of homeostatic plasticity in epilepsy. First, we will present some recent insights on how homeostatic compensations are probably active before and during epileptogenesis and how their actions are temporally regulated and closely dependent on the progression of pathology. Then, we will consider the dual role of transcriptional regulation during epileptogenesis, and finally, we will underline the importance of homeostatic plasticity in the context of therapeutic interventions for epilepsy. While classic pharmacological interventions may be counteracted by the epileptic brain to maintain its potentially dysfunctional set point, novel therapeutic approaches may provide the neuronal network with the tools necessary to restore its physiological balance., (Copyright © 2020 Lignani, Baldelli and Marra.)

Key Points: Kv3.1 and Kv3.3 subunits are highly expressed in the auditory brainstem, with little or no mRNA for Kv3.2 or Kv3.4. Changes in Kv3 currents and action potential (AP) firing were analysed from wild-type, Kv3.1 and Kv3.3 knockout (KO) mice. Both Kv3.1 and Kv3.3 immunostaining was present and western blots confirmed loss of subunit protein in the respective KO. Medial nucleus of the trapezoid body (MNTB) AP repolarization utilized Kv3.1 and/or Kv3.3; while in the lateral superior olive (LSO) Kv3.3 was essential. Voltage-gated calcium currents were unchanged between the genotypes. But APs evoked higher [Ca 2+ ] i in LSO than MNTB neurons; and were highest in the Kv3.3KO, consistent with longer AP durations. High frequency stimulation increased AP failure rates and AP latency in LSO neurons from the Kv3.3KO, underlining the physiological consequences for binaural integration. LSO neurons require Kv3.3 for functional Kv3 channels, while MNTB neurons can utilize either Kv3.1 or Kv3.3 subunits., Abstract: Kv3 voltage-gated potassium channels mediate action potential (AP) repolarization. The relative importance of Kv3.1 and Kv3.3 subunits for assembly of functional channels in neurons of the auditory brainstem was examined from the physiological perspective that speed and precision of AP firing are crucial for sound source localization. High levels of Kv3.1 and Kv3.3 mRNA and protein were measured, with no evidence of compensation by Kv3.2 or Kv3.4 in the respective knockout (KO) mouse. Using the KOs, composition of Kv3 channels was constrained to either Kv3.1 or Kv3.3 subunits in principal neurons of the medial nucleus of the trapezoid body (MNTB) and lateral superior olive (LSO); while TEA (1 mm) was employed to block Kv3-mediated outward potassium currents in voltage- and current clamp experiments. MNTB neuron APs (half-width 0.31 ± 0.08 ms, n = 25) were fast, reliable, and showed no distinction between channels assembled from Kv3.1 or Kv3.3 subunits (in the respective KO). LSO AP half-widths were also fast, but absolutely required Kv3.3 subunits for fast repolarization (half-widths: 0.25 ± 0.08 ms, n = 19 wild-type, 0.60 ± 0.17 ms, n = 21 Kv3.3KO, p = 0.0001). The longer AP duration increased LSO calcium influx and AP failure rates, and increased AP latency and jitter during high frequency repetitive firing. Both Kv3.1 and Kv3.3 subunits contribute to Kv3 channels in the MNTB (and compensate for each other in each KO); in contrast, LSO neurons require Kv3.3 subunits for fast repolarization and to sustain AP firing during high frequency stimulation. In conclusion, Kv3 channels exhibit both redundancy and Kv3.3 dominance between the brainstem nuclei involved in sound localization., (© 2020 The Authors. The Journal of Physiology published by John Wiley & Sons Ltd on behalf of The Physiological Society.)

Human exposure to per- and polyfluorinated alkyl substances (PFASs) is a major public health concern because in the last decades several cases of overexposure of people to PFASs, in particular through contaminated water, occurred worldwide. In 2013-2017 a PFAS drinking water contamination was discovered and investigated in northern Italy (Veneto region) and high PFAS serum levels were detected in exposed people. 629 subjects were enrolled: 257 residing in municipalities in the areas under impact, 250 residing in municipalities in areas at presumed background exposure and 122 farmers living in contaminated rural areas producing and consuming own livestock and vegetables and frequently using well water. The highest PFAS serum concentrations (median PFOA concentrations 40 ng/g) were found in the subgroup of farmers. The main factors influencing PFAS serum levels of farmers were residence area and the related extent of drinking water contamination, gender, years of residence in the municipalities, well water consumption and consumption of own produced food. PFOA serum concentrations in farmers residing in the areas of the Veneto region impacted by PFAS contamination are among the highest found worldwide., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Vesicle pool properties are known determinants of synaptic efficacy, but their potential role as modifiable substrates in forms of Hebbian plasticity is still unclear. Here, we investigate this using a nanoscale readout of functionally recycled vesicles in natively wired hippocampal CA3→CA1 circuits undergoing long-term potentiation (LTP). We show that the total recycled vesicle pool is larger after plasticity induction, with the smallest terminals exhibiting the greatest relative expansion. Changes in the spatial organization of vesicles accompany potentiation including a specific increase in the number of recycled vesicles at the active zone, consistent with an ultrastructural remodeling component of synaptic strengthening. The cAMP-PKA pathway activator, forskolin, selectively mimics some features of LTP-driven changes, suggesting that distinct and independent modules of regulation accompany plasticity expression. Our findings provide evidence for a presynaptic locus of LTP encoded in the number and arrangement of functionally recycled vesicles, with relevance for models of long-term plasticity storage., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)

Exposure to polycyclic aromatic hydrocarbons (PAHs) was assessed in a cohort of 394 subjects, 198 residing in three small municipalities near a new waste-to-energy (WTE) incinerator located in the Turin area, and 196 residing in neighbouring control areas in the town (of Turin). The assessment of exposure to PAHs was part of a human biomonitoring study aimed at assessing potential incremental exposure to pollutants related to incineration activities through the analysis of such pollutants before the plant start-up, and after one and three years of operation. The exposure assessment described in this study was carried out before the start-up of the WTE incinerator. Ten monohydroxy-PAHs (OH-PAHs) were analyzed in urine samples, consisting in the principal metabolites of naphthalene (NAP), fluorene (FLU), phenanthrene (PHE), and pyrene (PYR). Concentrations of the sum of OH-PAHs (Σ 10 OH-PAHs) were in the range of 525-85200 ng/g creatinine, with P 50 equal to 6770 ng/g creatinine. Metabolites of naphthalene were found at the highest concentrations (P 50 values of 892 and 4300 ng/g creatinine for 1- and 2-OH-NAP, respectively) followed by the three OH-FLUs (P 50 values of individual compounds in the range of 58.2-491 ng/g creatinine), the four OH-PHEs (P 50 values in the range of 30.5-145 ng/g creatinine), and 1-OH-PYR (P 50 value of 82.8 ng/g creatinine). Concentrations of 1-OH-NAP, 9-OH-FLU, 1-, 2-, 3, 4-OH-PHE, and 1-OH-PYR were significantly lower in subjects living near the WTE plant compared to those living in the town of Turin, with differences between the two groups in the range 14-31%. Smoking habits markedly influence the urinary concentrations OH-PAHs. Median concentrations of the single metabolites in smokers were from 1.4 fold (for 4-OH-PHE) to 14 fold higher (for 3-OH-FLU) than those observed in non-smokers. The heating system used also resulted to be a major contributor to PAH exposure. Concentrations of OH-PAHs were generally comparable with those observed in other industrialized countries. The profile pattern was consistent with those reported in the literature. Concentrations of OH-PAHs assessed in this study may be considered indicative of the background exposure to PAHs for adult population living in an urban and industrialized area., (Published by Elsevier Inc.)

Rumination, namely a cognitive process characterized by a repetitive thinking focused on negative feelings and thoughts, is a significant predictor for the onset of internalizing symptoms and has also been found to run in families. Rumination has never been studied in children with specific learning disorders (SLD), a population that, due to its condition, might encounter more difficulties in daily life and is at risk of increased psychological distress, compared to typically developing (TD) peers. The present study covers this gap by examining whether children with SLD, and their parents, tend to use rumination more than TD peers and their parents. The study also explores associations between rumination and both children's and parents' emotional profile. Results on 25 children with SLD and 25 TD peers and their parents ( n = 150), showed higher levels of rumination in children with SLD when referring to a negative social situation, as well as higher levels of rumination in both mothers and fathers of children with SLD. Modest correlations between parents' and children's rumination traits were also found. This study offers evidence on rumination as a possible risk factor for children with SLD, particularly considering when they deal with social contexts.

Here, we describe a cost-effective setup for targeted photoconversion of fluorescent signals into electron dense ones. This approach has offered invaluable insights in the morphology and function of fine neuronal structures. The technique relies on the localized oxidation of diaminobenzidine (DAB) mediated by excited fluorophores. This paper includes a detailed description of how to build a simple photoconversion setup that can increase reliability and throughput of this well-established technique. The system described here, is particularly well-suited for thick neuronal tissue, where light penetration and oxygen diffusion may be limiting DAB oxidation. To demonstrate the system, we use Correlative Light and Electron Microscopy (CLEM) to visualize functionally-labeled individual synaptic vesicles released onto an identified layer 5 neuron in an acute cortical slice. The setup significantly simplifies the photoconversion workflow, increasing the depth of photoillumination, improving the targeting of the region of interest and reducing the time required to process each individual sample. We have tested this setup extensively for the photoconversion of FM 1-43FX and Lucifer Yellow both excited at 473 nm. In principle, the system can be adapted to any dye or nanoparticle able to oxidize DAB when excited by a specific wavelength of light.

Key Points: Synapses have high energy demands which increase during intense activity. We show that presynaptic terminals can utilise extracellular glucose or lactate to generate energy to maintain synaptic transmission. Reducing energy substrates induces a metabolic stress: presynaptic ATP depletion impaired synaptic transmission through a reduction in the number of functional synaptic vesicle release sites and a slowing of vesicle pool replenishment, without a consistent change in release probability. Metabolic function is compromised in many pathological conditions (e.g. stroke, traumatic brain injury and neurodegeneration). Knowledge of how synaptic transmission is constrained by metabolic stress, especially during intense brain activity, will provide insights to improve cognition following pathological insults., Abstract: The synapse has high energy demands, which increase during intense activity. Presynaptic ATP production depends on substrate availability and usage will increase during activity, which in turn could influence transmitter release and information transmission. We investigated transmitter release at the mouse calyx of Held synapse using glucose or lactate (10, 1 or 0 mm) as the extracellular substrates while inducing metabolic stress. High-frequency stimulation (HFS) and recovery paradigms evoked trains of EPSCs monitored under voltage-clamp. Whilst postsynaptic intracellular ATP was stabilised by diffusion from the patch pipette, depletion of glucose increased EPSC depression during HFS and impaired subsequent recovery. Computational modelling of these data demonstrated a reduction in the number of functional release sites and slowed vesicle pool replenishment during metabolic stress, with little change in release probability. Directly depleting presynaptic terminal ATP impaired transmitter release in an analogous manner to glucose depletion. In the absence of glucose, presynaptic terminal metabolism could utilise lactate from the aCSF and this was blocked by inhibition of monocarboxylate transporters (MCTs). MCT inhibitors significantly suppressed transmission in low glucose, implying that lactate is a presynaptic substrate. Additionally, block of glycogenolysis accelerated synaptic transmission failure in the absence of extracellular glucose, consistent with supplemental supply of lactate by local astrocytes. We conclude that both glucose and lactate support presynaptic metabolism and that limited availability, exacerbated by high-intensity firing, constrains presynaptic ATP, impeding transmission through a reduction in functional presynaptic release sites as vesicle recycling slows when ATP levels are low., (© 2018 The Authors. The Journal of Physiology © 2018 The Physiological Society.)

Euclid is a European Space Agency medium-class mission selected for launch in 2020 within the cosmic vision 2015-2025 program. The main goal of Euclid is to understand the origin of the accelerated expansion of the universe. Euclid will explore the expansion history of the universe and the evolution of cosmic structures by measuring shapes and red-shifts of galaxies as well as the distribution of clusters of galaxies over a large fraction of the sky. Although the main driver for Euclid is the nature of dark energy, Euclid science covers a vast range of topics, from cosmology to galaxy evolution to planetary research. In this review we focus on cosmology and fundamental physics, with a strong emphasis on science beyond the current standard models. We discuss five broad topics: dark energy and modified gravity, dark matter, initial conditions, basic assumptions and questions of methodology in the data analysis. This review has been planned and carried out within Euclid's Theory Working Group and is meant to provide a guide to the scientific themes that will underlie the activity of the group during the preparation of the Euclid mission.

In 2013 a contamination of drinking water by perfluoroalkylated substances (PFASs) was discovered in areas of the Veneto Region (northern Italy). In this study the exposure to PFASs of people living in the aforesaid areas was characterized: contaminant serum concentrations were measured and compared with those of a control population group living in neighboring areas at background exposure (based on available drinking water data). The enrolled population was also genotyped for the OATP1A2*3 allelic variant, possibly affecting PFAS excretion and hence the internal dose. The difference in PFAS concentrations between exposed and not exposed subjects was significantly larger for nine of the 12 substances analyzed, and confirmed that water contamination had resulted in an appreciable high exposure of the residing population over time. Within the group of exposed subjects, subgroups at different exposure levels were identified. The contamination of drinking water of the residence area was found to be the main factor influencing PFAS serum levels; in addition to water contamination, other relevant influencing factors were sex, the years of residence and raising own livestock. No relationship with the genetic trait for the studied renal transporter was evidenced. These results provide a baseline characterization of PFAS exposure of the monitored population groups for further studies, planned to be carried out in the near future., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Background: Polychlorinated dibenzodioxins (PCDDs), polychlorodibenzofurans (PCDFs), and polychlorobiphenyls (PCBs) are persistent organic pollutants that represent a major concern for women of reproductive age because of the neurodevelopmental effects associated to perinatal exposure., Objectives: This study was aimed at characterizing exposure of women of reproductive age to PCDDs, PCDFs, and PCBs as a function of residence in different Italian Regions, in areas at presumable different environmental contamination and human exposure to these pollutants., Methods: Study participants were enrolled in 2011-2012 in 6 Italian Regions representative of Northern, Central and Southern Italy; in each region, areas at presumed different exposure (rural, urban and industrial) were selected for enrolment. Each participant provided a serum sample for the analysis of PCDDs, PCDFs and PCBs., Results: Median concentrations of PCDDs+PCDFs, DL-PCBs, NDL 6 -PCBs and NDL 9 -PCBs in serum samples were respectively 6.0 and 3.5 pgWHO-TE 05 /g fat, and 75 and 93ng/g fat. Age was the variable that most affected median serum concentrations. Age adjusted concentrations were found significantly different between geographical zones: women from Northern Italy showed the highest values, followed by Central and Southern Italy. PCDDs+PCDFs concentrations were significantly higher in the group of women residing in industrial areas compared to the group residing in rural areas. A clear diminishing temporal trend was observed compared to levels reported in previous studies., Conclusions: This study produced the largest dataset on serum concentrations of PCDDs, PCDFs and PCBs in women of childbearing age in Italy., Results: confirmed that environmental and lifestyle factors may influence exposure to these contaminants and thereby the body burden. The observed marked temporal decline in body burden during three decades is in agreement with the general trend observed worldwide., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

Objectives: To compare breast density (BD) assessment provided by an automated BD evaluator (ABDE) with that provided by a panel of experienced breast radiologists, on a multivendor dataset., Methods: Twenty-one radiologists assessed 613 screening/diagnostic digital mammograms from nine centers and six different vendors, using the BI-RADS a, b, c, and d density classification. The same mammograms were also evaluated by an ABDE providing the ratio between fibroglandular and total breast area on a continuous scale and, automatically, the BI-RADS score. A panel majority report (PMR) was used as reference standard. Agreement (κ) and accuracy (proportion of cases correctly classified) were calculated for binary (BI-RADS a-b versus c-d) and 4-class classification., Results: While the agreement of individual radiologists with the PMR ranged from κ = 0.483 to κ = 0.885, the ABDE correctly classified 563/613 mammograms (92 %). A substantial agreement for binary classification was found for individual reader pairs (κ = 0.620, standard deviation [SD] = 0.140), individual versus PMR (κ = 0.736, SD = 0.117), and individual versus ABDE (κ = 0.674, SD = 0.095). Agreement between ABDE and PMR was almost perfect (κ = 0.831)., Conclusions: The ABDE showed an almost perfect agreement with a 21-radiologist panel in binary BD classification on a multivendor dataset, earning a chance as a reproducible alternative to visual evaluation., Key Points: Individual BD assessment differs from PMR with κ as low as 0.483. An ABDE correctly classified 92 % of mammograms with almost perfect agreement (κ = 0.831). An ABDE can be a valid alternative to subjective BD assessment.

In the mammalian brain, synaptic transmission usually depends on presynaptic action potentials (APs) in an all-or-none (or digital) manner. Recent studies suggest, however, that subthreshold depolarization in the presynaptic cell facilitates spike-evoked transmission, thus creating an analogue modulation of a digital process (or analogue-digital (AD) modulation). At most synapses, this process is slow and not ideally suited for the fast dynamics of neural networks. We show here that transmission at CA3-CA3 and L5-L5 synapses can be enhanced by brief presynaptic hyperpolarization such as an inhibitory postsynaptic potential (IPSP). Using dual soma-axon patch recordings and live imaging, we find that this hyperpolarization-induced AD facilitation (h-ADF) is due to the recovery from inactivation of Nav channels controlling AP amplitude in the axon. Incorporated in a network model, h-ADF promotes both pyramidal cell synchrony and gamma oscillations. In conclusion, cortical excitatory synapses in local circuits display hyperpolarization-induced facilitation of spike-evoked synaptic transmission that promotes network synchrony.

Purpose: To evaluate a commercial tomosynthesis computer-aided detection (CAD) system in an independent, multicenter dataset., Materials and Methods: Diagnostic and screening tomosynthesis mammographic examinations (n = 175; cranial caudal and mediolateral oblique) were randomly selected from a previous institutional review board-approved trial. All subjects gave informed consent. Examinations were performed in three centers and included 123 patients, with 132 biopsy-proven screening-detected cancers, and 52 examinations with negative results at 1-year follow-up. One hundred eleven lesions were masses and/or microcalcifications (72 masses, 22 microcalcifications, 17 masses with microcalcifications) and 21 were architectural distortions. Lesions were annotated by radiologists who were aware of all available reports. CAD performance was assessed as per-lesion sensitivity and false-positive results per volume in patients with negative results., Results: Use of the CAD system showed per-lesion sensitivity of 89% (99 of 111; 95% confidence interval: 81%, 94%), with 2.7 ± 1.8 false-positive rate per view, 62 of 72 lesions detected were masses, 20 of 22 were microcalcification clusters, and 17 of 17 were masses with microcalcifications. Overall, 37 of 39 microcalcification clusters (95% sensitivity, 95% confidence interval: 81%, 99%) and 79 of 89 masses (89% sensitivity, 95% confidence interval: 80%, 94%) were detected with the CAD system. On average, 0.5 false-positive rate per view were microcalcification clusters, 2.1 were masses, and 0.1 were masses and microcalcifications., Conclusion: A digital breast tomosynthesis CAD system can allow detection of a large percentage (89%, 99 of 111) of breast cancers manifesting as masses and microcalcification clusters, with an acceptable false-positive rate (2.7 per breast view). Further studies with larger datasets acquired with equipment from multiple vendors are needed to replicate the findings and to study the interaction of radiologists and CAD systems., ((©) RSNA, 2015.)

Perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) concentrations were determined in serum samples collected in 2011-2012 from 549 nulliparous Italian women of reproductive age who resided in six different Italian Regions. Assessment of exposure to perfluorinated compounds was part of a large human biomonitoring study (Project Life Plus "Womenbiopop") that aimed at examining the exposure of women of reproductive age to priority organic pollutants. The median concentrations of PFOS and PFOA were 2.43, and 1.55 ng g(-1), respectively. Significant differences in the concentrations of both compounds were observed among the six Regions. Women from central Italy had the highest levels of both compounds, followed by women from northern Italy, and southern Italy. No differences in the PFOS concentrations were found between women from urban/industrial areas and women from rural areas, whereas the levels of PFOA were significantly higher in women residing in urban/industrial areas than in women residing in rural areas. Taken together, the observed concentrations confirm that the overall exposure of the Italian population is among the lowest observed in industrialized countries. A downward temporal trend in exposure was observed for both compounds when comparing the results from the present study with those assessed in a study conducted in 2008., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Synaptic transmission usually depends on action potentials (APs) in an all-or-none (digital) fashion. Recent studies indicate, however, that subthreshold presynaptic depolarization may facilitate spike-evoked transmission, thus creating an analog modulation of spike-evoked synaptic transmission, also called analog-digital (AD) synaptic facilitation. Yet, the underlying mechanisms behind this facilitation remain unclear. We show here that AD facilitation at rat CA3-CA3 synapses is time-dependent and requires long presynaptic depolarization (5-10 s) for its induction. This depolarization-induced AD facilitation (d-ADF) is blocked by the specific Kv1.1 channel blocker dendrotoxin-K. Using fast voltage-imaging of the axon, we show that somatic depolarization used for induction of d-ADF broadened the AP in the axon through inactivation of Kv1.1 channels. Somatic depolarization enhanced spike-evoked calcium signals in presynaptic terminals, but not basal calcium. In conclusion, axonal Kv1.1 channels determine glutamate release in CA3 neurons in a time-dependent manner through the control of the presynaptic spike waveform., (© 2014 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.)

Fast activity-driven turnover of neurotransmitter-filled vesicles at presynaptic terminals is a crucial step in information transfer in the CNS. Characterization of the relationship between the nanoscale organization of synaptic vesicles and their functional properties during transmission is currently of interest. Here we outline a procedure for ultrastructural investigation of functional vesicles in synapses from native mammalian brain tissue. FM dye is injected into the target region of a brain slice and upstream axons are electrically activated to stimulate vesicle turnover and dye uptake. In the presence of diaminobenzidine (DAB), photoactivation of dye-filled vesicles yields an osmiophilic precipitate that is visible in electron micrographs. When combined with serial-section electron microscopy, fundamental ultrastructure-function relationships of presynaptic terminals in native circuits are revealed. We outline the utility of this protocol for the 3D reconstruction of a recycling vesicle pool in CA3-CA1 synapses from an acute hippocampal slice and for the characterization of its anatomically defined docked pool. This protocol requires 6-7 d.

There is an approximately 9% discrepancy, corresponding to 2.4 σ, between two independent constraints on the expansion rate of the Universe: one indirectly arising from the cosmic microwave background and baryon acoustic oscillations and one more directly obtained from local measurements of the relation between redshifts and distances to sources. We argue that by taking into account the local gravitational potential at the position of the observer this tension--strengthened by the recent Planck results--is partially relieved and the concordance of the Standard Model of cosmology increased. We estimate that measurements of the local Hubble constant are subject to a cosmic variance of about 2.4% (limiting the local sample to redshifts z > 0.010) or 1.3% (limiting it to z > 0.023), a more significant correction than that taken into account already. Nonetheless, we show that one would need a very rare fluctuation to fully explain the offset in the Hubble rates. If this tension is further strengthened, a cosmology beyond the Standard Model may prove necessary.

The aim of this study was to characterize the placental transfer of some environmental pollutants, and to explore the possibility of quantitatively predicting in utero exposure to these contaminants from concentrations assessed in maternal blood. Levels of toxic substances such as pesticides (p,p'-DDE, β-HCH, and HCB), polychlorinated biphenyls (PCBs), perfluorooctane sulfonate (PFOS), and perfluorooctanoic acid (PFOA) were determined in serum samples of 38 pregnant women living in Rome and in samples of cord blood from their respective newborns. The study was carried out in the years 2008-2009. PCB mean concentrations in maternal serum and cord serum ranged from 0.058 to 0.30, and from 0.018 to 0.064 ng/g · fw respectively. Arithmetic means of PFOS and PFOA concentrations in mothers and newborns were 3.2 and 1.4 ng/g · fw, and 2.9 and 1.6 ng/g · fw. A strong correlation was observed between concentrations in the maternal and the foetal compartment for PFOS (Spearman r = 0.74, p < 0.001), PFOA (Spearman r = 0.70, p < 0.001), PCB 153 (Spearman r = 0.60, p < 0.001), HCB (Spearman r = 0.68, p < 0.001), PCB 180 (Spearman r = 0.55, p = 0.0012), and p,p'-DDE (Spearman r = 0.53, p = 0.0099). A weak correlation (p < 0.1) was observed for PCBs 118 and 138.

Background: The study was carried out in order to respond to public concern on the occupational exposure of metallurgical workers to highly toxic PCDDs, PCDFs, and PCBs in the area of the city of Brescia, northern Italy., Objectives: The study investigated the effects on the haematic burden of occupational exposures to the aforesaid contaminants in different work environments, attempting to establish causal relationships and providing indications for occupational health preventive measures., Methods: Chemical concentrations were measured in blood serum of "professionally exposed" (PE) and "not professionally exposed" (NPE) subjects. NPE subjects included industrial administrative employees, Brescia inhabitants, and remote rural people., Results: The central tendency indexes of contaminant cumulative concentrations were higher in PE than in NPE samples (for the mean values: PCDDs+PCDFs, 22.9 vs. 19.5 pgWHO-TEQ(1997)/g lb; DL-PCBs, 26.0 vs. 23.6 pgWHO-TEQ(1997)/g lb; PCDDs+PCDFs+DL-PCBs (TEQ(TOT)), 48.9 vs. 43.1 pgWHO-TEQ(1997)/g lb; Σ(6)[NDL-PCBs], 427 vs. 401 ng g(-1)lb); however, no statistical differences were detected at P=0.05. A significant difference for PCDDs+PCDFs and TEQ(TOT) was observed as the NPE data were progressively reduced to those of the remote rural people. The existence of a differential occupational exposure due to different environments was detected by applying the factor analysis to congener-specific data (analytical profiles)., Conclusions: Findings indicate that metallurgical workers may be exposed to PCDD, PCDF, and PCB more than the general population, in particular due to non-negligible contributions to exposure from workplace ambient air. Findings also suggest that an improvement of preventive measures may be required to avoid chemical overexposure in certain metallurgical workplaces. To identify exposure groups, the DL- and NDL-PCB analytical profiles seemed to be more sensitive to environmental exposure sources/pathways than those of PCDDs and PCDFs., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Objectives: To describe the pitfalls encountered in switching over from conventional smears (CSm) to liquid-based cytology (LBC). To explore modifications of our usual diagnostic criteria., Study Design: 190 ThinPrep breast samples with paired biopsies were retrospectively evaluated by two breast cytopathologists experienced only in CSm. They were again studied after LBC training. The diagnostic performances were compared. In additional calculations we included those C4/suspicious samples containing 70% high-grade nuclei and up to 30% nonmalignant cells in the C5/positive category, simulating that they harbored a malignant one-cell population. We prospectively validated this modification of our diagnostic criteria., Results: Training resulted in higher complete sensitivity: 94 versus 86% (p value 0.003) and lower false negative ratio: 4 versus 12% (p value 0.003). Training generated higher complete sensitivity than collaboration without training: 94 versus 89% (p value 0.008). In the simulation, the modified criteria increased absolute sensitivity to 74% with a 0.6% false positive rate. In the validation series, they generated up to 91% absolute sensitivity, 12% suspicious rate and no false negative and false positive diagnoses., Conclusion: Training in breast LBC may increase diagnostic performance. Samples containing 70% high-grade nuclei or more can be categorized as malignant., (Copyright © 2013 S. Karger AG, Basel.)

Memories that can be recalled several hours after learning may paradoxically become inaccessible for brief periods after their formation. This raises major questions about the function of these early memory lapses in the structure of memory consolidation. These questions are difficult to investigate because of the lack of information on the precise timing of lapses. However, the use of a single-trial conditioning paradigm in Lymnaea solves this problem. Here we use electrophysiological and behavioural experiments to reveal lapses in memory recall at 30 min and 2 h post conditioning. We show that only during these lapses is consolidation of long-term memory susceptible to interruption by external disturbance. These shared time points of memory lapse and susceptibility correspond to transitions between different phases of memory that have different molecular requirements. We propose that during periods of molecular transition memory recall is weakened, allowing novel sensory cues to block the consolidation of long-term memory.

At small central synapses, efficient turnover of vesicles is crucial for stimulus-driven transmission, but how the structure of this recycling pool relates to its functional role remains unclear. Here we characterize the organizational principles of functional vesicles at native hippocampal synapses with nanoscale resolution using fluorescent dye labeling and electron microscopy. We show that the recycling pool broadly scales with the magnitude of the total vesicle pool, but its average size is small (∼45 vesicles), highly variable, and regulated by CDK5/calcineurin activity. Spatial analysis demonstrates that recycling vesicles are preferentially arranged near the active zone and this segregation is abolished by actin stabilization, slowing the rate of activity-driven exocytosis. Our approach reveals a similarly biased recycling pool distribution at synapses in visual cortex activated by sensory stimulation in vivo. We suggest that in small native central synapses, efficient release of a limited pool of vesicles relies on their favored spatial positioning within the terminal., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Objective: to conduct a survey on blood levels of metals and organochlorine compounds (dioxins and PCBs) among workers of livestock farms, and to identify the factors able to explain the magnitude and variability of the measured concentrations., Design: exploratory human biomonitoring study., Setting and Participants: a sample of 45 workers from farms (masserie) located in the Province of Taranto was included in the study. The farms, located between 6 and 43 km from the industrial area, are family owned and animals (mainly sheep and goats) are locally reared outdoors. The study subjects have similar eating habits, including food consumption resulting from their production., Results: manganese concentrations and, to a lesser extent, arsenic, cadmium and lead are in the medium-high range of levels observed in the Italian general population. This study does not identify factors able to explain the variability in the blood levels of metals. The amount of dioxins and PCBs, on the other hand, is strongly associated both with subjects age and the distance of farm from the industrial site., Conclusion: the results of this survey are suggestive of research hypotheses that need to be confirmed by biomonitoring studies of adequate design and size, in particular the hypothesis that the blood level of dioxins and PCBs decreases with increasing distance from the industrial site. We recommend ad hoc studies to better characterize the exposure of farmers (also associated with agricultural activities), and of the inhabitants of Taranto, to environmental persistent pollutants, also in terms of food matrices locally produced and consumed.

Dynamic clamp is a powerful approach for electrophysiological investigations allowing researchers to introduce artificial electrical components into target neurons to simulate ionic conductances, chemical or electrotonic inputs or connections to other cells. Due to the rapidly changing and potentially large current injections during dynamic clamp, problematic voltage artifacts appear on the electrode used to inject dynamic clamp currents into a target neuron. Dynamic clamp experiments, therefore, typically use two separate electrodes in the same cell, one for recording membrane potential and one for injecting currents. The requirement for two independent electrodes has been a limiting factor for the use of dynamic clamp in applications where dual recordings of this kind are difficult or impossible to achieve. The recent development of an active electrode compensation (AEC) method has overcome some of these prior limitations, permitting artifact-free dynamic clamp experimentation with a single electrode. Here we describe an AEC method for the free dynamic clamp software StdpC. The AEC component of StdpC is the first such system implemented for the use of non-expert users and comes with a set of semi-automated configuration and calibration procedures that facilitate its use. We briefly introduce the AEC method and its implementation in StdpC and then validate it with an electronic model cell and in two different biological preparations., (Copyright © 2012 Elsevier B.V. All rights reserved.)

High temperature metallurgical processes may significantly contribute to the release of polychlorodibenzo-p-dioxins (PCDDs) and polychlorodibenzofurans (PCDFs) into the environment. Aim of this study was to evaluate the exposure to PCDDs, PCDFs, dioxin-like PCBs (DL-PCBs) and non dioxin-like PCBs (NDL-PCBs) in a sample of 300 workers of metallurgical plants operating in the Brescia area and in a control group of 113 men. Serum samples were grouped in pools of 10 and analyzed for their concentration of PCDDs and PCDFs, DL- and NDL-PCBs. Metallurgical workers show serum concentrations of NDL-PCBs similar to those observed in non-occupationally exposed living in Brescia, except in the case of some congeners, such as PCBs 28, 52, 101 which result to be significantly higher in metallurgical workers. Their concentrations range of "dioxins" was of 32.5 - 84.8 pgTEQ g(-1) lb. No significant difference was found between metallurgical workers and the control group on the whole, but the difference became significant when comparing metallurgical workers with the group of the 47 subjects living in remote non industrialised areas. In conclusion, concentrations of total PCBs observed in metallurgy workers are comparable with those observed in the control group, with the exception of a few, more volatile, congeners which resulted to be higher in metallurgy workers. Concentrations of PCDDs and PCDFs showed a range of values similar to that observed in the general population of Brescia not professionally exposed. However, concentrations of PCDDs and PCDFs observed in subjects resident in "remote" areas were significantly lower than those assessed in metallurgy workers and subjects living in the town of Brescia. On the whole, men living in the town of Brescia show levels of PCBs, PCDDs and PCDFs higher than those observed in other groups of the Italian general population, this reflecting the intense industrial activity of the town.

Background: Several factors can influence access to population breast cancer screening. The aim of the study was to evaluate the impact of different information approaches, women's socio-demographic characteristics and organizational factors on mammography screening uptake., Methods: We selected 5744 women aged 40-45 years who were randomly assigned to be given letters with: (i) a pre-fixed appointment plus standard leaflet (Group 1); (ii) a pre-fixed appointment plus a more comprehensive booklet (Group 2); (iii) point (ii) plus the offer of a counselling session (Group 3); and (iv) an invitation to contact the centre to get information and arrange participation (Group 4)., Results: Ninety-five women were excluded before the invitation and 5649 were randomized. After excluding undelivered letters (n = 41) and women reporting an exclusion criterion following our invitation (n = 248), the final eligible population was 5360 women. Participation rates following the first contact were 36.5, 39.9, 35.8 and 16.5% for Groups 1-4, respectively. The rates increased to 40.9, 43.6, 40.1 and 35.1% after the reminder letters. Women receiving more complete information had a higher uptake (Group 2), although not statistically significant. Differences among the four groups were maintained by controlling the effect of socio-demographic and attendance determinants. Regardless of intervention, participation was higher among married, higher educated, white-collared women, those born in northern Italy, living closer to the screening unit and with a female-collaborative doctor., Conclusion: Invitation letters with a fixed appointment correlate with a higher attendance rate. Providing women with more information on procedures, risks and benefits of mammography screening does not modify their participation.

Most presynaptic terminals in the central nervous system are characterized by two functionally distinct vesicle populations: a recycling pool, which supports action potential-driven neurotransmitter release via vesicle exocytosis, and a resting pool. The relative proportions of these two pools are highly variable between individual synapses, prompting speculation on their specific relationship, and on the possible functions of the resting pool.Using fluorescence imaging of FM-styryl dyes and synaptophysinI-pHluorin(sypHy) as well as correlative electronmicroscopy approaches, we show here that Hebbian plasticity-dependent changes in synaptic strength in rat hippocampal neurons can increase the recycling pool fraction at the expense of the resting pool in individual synaptic terminals. This recruitment process depends on NMDA-receptor activation, nitric oxide signalling and calcineurin and is accompanied by an increase in the probability of neurotransmitter release at individual terminals. Blockade of actin-mediated intersynaptic vesicle exchange does not prevent recycling pool expansion demonstrating that vesicle recruitment is intrasynaptic.We propose that the conversion of resting pool vesicles to the functionally recycling pool provides a rapid mechanism to implement long-lasting changes in presynaptic efficacy.

Fast neuronal signalling relies on highly regulated vesicle fusion and recycling at specialized presynaptic terminals. Recently, examples of non-classical neurotransmission have also been reported, where fusion of vesicles can occur at sites remote from conventional synapses. This has potentially broad biological implications, but the underlying mechanisms are not well established. Here we show that a complete vesicle recycling pathway can occur at discrete axonal sites in mature hippocampal neurons and that extrasynaptic fusion is a robust feature of native tissue. We demonstrate that laterally mobile vesicle clusters trafficking between synaptic terminals become transiently stabilized by evoked action potentials and undergo complete but delayed Ca(2+)-dependent fusion along axons. This fusion is associated with dynamic actin accumulation and, subsequently, vesicles can be locally recycled, re-acidified and re-used. Immunofluorescence and ultrastructural work demonstrates that extrasynaptic fusion sites can have apposed postsynaptic specializations, suggesting that mobile vesicle recycling may underlie highly dynamic neuron-neuron communication.

Virus infections pose significant global health challenges, especially in view of the fact that the emergence of resistant viral strains and the adverse side effects associated with prolonged use continue to slow down the application of effective antiviral therapies. This makes imperative the need for the development of safe and potent alternatives to conventional antiviral drugs. In the present scenario, nanoscale materials have emerged as novel antiviral agents for the possibilities offered by their unique chemical and physical properties. Silver nanoparticles have mainly been studied for their antimicrobial potential against bacteria, but have also proven to be active against several types of viruses including human imunodeficiency virus, hepatitis B virus, herpes simplex virus, respiratory syncytial virus, and monkey pox virus. The use of metal nanoparticles provides an interesting opportunity for novel antiviral therapies. Since metals may attack a broad range of targets in the virus there is a lower possibility to develop resistance as compared to conventional antivirals. The present review focuses on the development of methods for the production of silver nanoparticles and on their use as antiviral therapeutics against pathogenic viruses.

Dynamic clamp is a powerful method that allows the introduction of artificial electrical components into target cells to simulate ionic conductances and synaptic inputs. This method is based on a fast cycle of measuring the membrane potential of a cell, calculating the current of a desired simulated component using an appropriate model and injecting this current into the cell. Here we present a dynamic clamp protocol using free, fully integrated, open-source software (StdpC, for spike timing-dependent plasticity clamp). Use of this protocol does not require specialist hardware, costly commercial software, experience in real-time operating systems or a strong programming background. The software enables the configuration and operation of a wide range of complex and fully automated dynamic clamp experiments through an intuitive and powerful interface with a minimal initial lead time of a few hours. After initial configuration, experimental results can be generated within minutes of establishing cell recording.

Release probability (p(r)) is a fundamental presynaptic parameter which is critical in defining synaptic strength. Knowledge of how synapses set and regulate their p(r) is a fundamental step in understanding synaptic transmission and communication between neurons. Despite its importance, p(r) is difficult to measure directly at single synapses. One important strategy to achieve this has relied on the application of fluorescence-based imaging methods, but this is always limited by the lack of detailed information on the morphological and structural properties of the individual synapses under study, and thus precludes an investigation of the relationship between p(r) and synaptic anatomy. Here we outline a powerful methodology based on using FM-styryl dyes, photoconversion and correlative ultrastructural analysis in dissociated hippocampal cultured neurons, which provides both a direct readout of p(r) as well as nanoscale detail on synaptic organization and structure. We illustrate the value of this approach by investigating, at the level of individual reconstructed terminals, the relationship between release probability and defined vesicle pools. We show that in our population of synapses, p(r) is highly variable, and while it is positively correlated with the number of vesicles docked at the active zone it shows no relationship with the total number of synaptic vesicles. The lack of a direct correlation between total synaptic size and performance in these terminals suggests that factors other than the absolute magnitude of the synapse are the most important determinants of synaptic efficacy., (Copyright © 2009 Elsevier Inc. All rights reserved.)

We reanalyze the supernova data from the Union Compilation including the weak-lensing effects caused by inhomogeneities. We compute the lensing probability distribution function for each background solution described by the parameters Ω(M), Ω(Λ), and w in the presence of inhomogeneities, approximately modeled with a single-mass population of halos. We then perform a likelihood analysis in the parameter space of Friedmann-Lemaître-Robertson-Walker models and compare our results with the standard approach. We find that the inclusion of lensing can move the best-fit model significantly towards the cosmic concordance of the flat Lambda-Cold Dark Matter model, improving the agreement with the constraints coming from the cosmic microwave background and baryon acoustic oscillations.

Changes in the strength of excitatory synaptic connections are known to underlie associative memory formation in the molluscan nervous system but less is known about the role of synaptic inhibition. Tonic or maintained synaptic inhibition has an important function in controlling the Lymnaea feeding system and is known to suppress feeding in the absence of food or in satiated animals. Tonic inhibition to the feeding network is provided by the N3t interneuron that has inhibitory monosynaptic connection with the central pattern generator interneuron, the N1M. Here we asked whether a reduction in the level of tonic inhibition provided by the N3t cell could play a role in reward conditioning? Semi-intact preparations made from hungry snails were conditioned using a previously developed one-trial chemical conditioning paradigm. We recorded electrical activity in a feeding motoneuron, the B3, at various time-points after conditioning. This allowed us to measure the frequency of spike activity in the N3t interneuron and monitor fictive feeding patterns that generate the rhythmic movements involved in food ingestion. We show that there is a reduction in N3t spiking at 1, 2, 3, and 4 h after conditioning but not at 10 and 30 min and the reduction in N3t firing inversely correlates with an increase in the conditioned fictive feeding response. Computer simulation of N3t-N1M interactions suggests that changes in N3t firing are sufficient to explain the increase in the fictive feeding activity produced by conditioning. A network model is presented that summarizes evidence suggesting that reward conditioning in Lymnaea is due to the combined effects of reduced tonic inhibition and enhanced excitatory synaptic connections between the CS pathway and feeding command neurons.

The serum concentrations of perfluorooctanesulfonate (PFOS) and perfluorooctanoic acid (PFOA) were determined in 230 subjects of the Italian general population. Participants were enrolled in 2008 in two Italian towns (Brescia, Northern Italy, and Rome, Central Italy) and belonged to the three age ranges: 20-35 years, 36-50 years, and 51-65 years. PFOS and PFOA were quantified by HPLC interfaced to a mass spectrometer operating in the electrospray negative mode. Data were acquired using multiple reaction monitoring (MRM). The isotope dilution technique was applied throughout. The median serum concentrations of all participants were 6.31 ng g(-1) and 3.59 ng g(-1) for PFOS and PFOA, respectively, and the pertinent 90th percentiles were 12.38 and 6.92. Men had higher concentrations of PFOS and PFOA than women, regardless of age. The differences were statistically significant in the 20-35 and 36-50 years groups, but not in the 51-65 group. An increase of PFOS and PFOA serum concentrations with age was observed. The Median test showed a statistically significant difference (p<<0.01) between the three age groups for both PFOS and PFOA when applied to the entire dataset (males and females). When the test was applied to the groups of males and females separately, a significant difference was observed for females (p<<0.005) but not for males (p>0.1). The observed strong correlation between PFOS and PFOA concentrations suggests same or similar exposure routes., (Copyright (c) 2010 Elsevier Ltd. All rights reserved.)

The Cerebral Giant Cells (CGCs) are a pair of identified modulatory interneurons in the Central Nervous System of the pond snail Lymnaea stagnalis with an important role in the expression of both unconditioned and conditioned feeding behavior. Following single-trial food-reward classical conditioning, the membrane potential of the CGCs becomes persistently depolarized. This depolarization contributes to the conditioned response by facilitating sensory cell to command neuron synapses, which results in the activation of the feeding network by the conditioned stimulus. Despite the depolarization of the membrane potential, which enables the CGGs to play a key role in learning-induced network plasticity, there is no persistent change in the tonic firing rate or shape of the action potentials, allowing these neurons to retain their normal network function in feeding. In order to understand the ionic mechanisms of this novel combination of plasticity and stability of intrinsic electrical properties, we first constructed and validated a Hodgkin-Huxley-type model of the CGCs. We then used this model to elucidate how learning-induced changes in a somal persistent sodium and a delayed rectifier potassium current lead to a persistent depolarization of the CGCs whilst maintaining their firing rate. Including in the model an additional increase in the conductance of a high-voltage-activated calcium current allowed the spike amplitude and spike duration also to be maintained after conditioning. We conclude therefore that a balanced increase in three identified conductances is sufficient to explain the electrophysiological changes found in the CGCs after classical conditioning.

Synapse-specific vesicle pools have been widely characterized at central terminals. Here, we demonstrate a vesicle pool that is not confined to a synapse but spans multiple terminals. Using fluorescence imaging, correlative electron microscopy, and modeling of vesicle dynamics, we show that some recycling pool vesicles at synapses form part of a larger vesicle "superpool." The vesicles within this superpool are highly mobile and are rapidly exchanged between terminals (turnover: approximately 4% of total pool/min), significantly changing vesicular composition at synapses over time. In acute hippocampal slices we show that the mobile vesicle pool is also a feature of native brain tissue. We also demonstrate that superpool vesicles are available to synapses during stimulation, providing an extension of the classical recycling pool. Experiments using focal BDNF application suggest the involvement of a local TrkB-receptor-dependent mechanism for synapse-specific regulation of presynaptic vesicle pools through control of vesicle release and capture to or from the extrasynaptic pool., (Copyright 2010 Elsevier Inc. All rights reserved.)

Background: Prenatal exposures to endocrine-disrupting chemicals (EDCs) are suspected risk factors in the etiology of hypospadias. The aim of this case-control study was to test the hypothesis of an association between maternal environmental exposures to EDCs and hypospadias in the offspring., Methods: Detailed questionnaire data on occupational and dietary exposures to EDCs in the perinatal period were collected from 80 mothers with hypospadiac infants and from 80 mothers with healthy controls within 24 months of childbirth. Maternal exposure to selected EDCs was also ascertained by measuring the concentration of dichlorodiphenyldichloroethylene, hexachlorobenzene, and several polychlorinated biphenyl congeners in the serum of primiparous mothers of 37 cases and 21 controls., Results: The risk to bear an hypospadiac infant was associated with perinatal maternal occupational exposures to EDCs evaluated by a job-exposure matrix: jobs with exposure to one class of EDCs (odds ratios [OR](crude), 2.83; 95% confidence intervals [CI], 1.32-6.07; OR(adjusted), 2.44; 95% CI, 1.06-5.61) and jobs with exposure to more than one group of EDCs (OR(crude), 4.27; 95% CI, 1.43-12.78; OR(adjusted), 4.11; 95%CI, 1.34-12.59). Increase in risk was also found among mothers consuming a diet rich in fish or shellfish (OR(crude), 3.41; 95% CI, 1.42-8.23; OR(adjusted), 2.73; 95%CI, 1.09-6.82). Serum hexachlorobenzene concentration above the median of all subjects was significantly associated with the risk of hypospadias (OR(adjusted), 5.50; 95% CI, 1.24-24.31)., Conclusions: This study, although based on a limited number of cases, for the first time provides evidence of an association between maternal exposure to EDCs, in particular elevated plasma hexachlorobenzene concentration, and the development of hypospadias in the offspring., ((c) 2010 Wiley-Liss, Inc.)

Because of its persistence and toxicological profile, beta-hexachlorocyclohexane (beta-HCH) has been proposed for inclusion in the Stockholm Convention on persistent organic pollutants (POPs). Although the use of technical HCH, which is the primary source of beta-HCH in the environment, has been banned in the EU in 1978 and progressively at a global level, beta-HCH is still detectable in the general environment worldwide. Human exposure mostly occurs via food and may be of concern in areas where illegal use and/or improper disposal of stockpiles occurred and locally grown food is consumed. Exposure of the Italian general population to beta-HCH has been poorly characterised. Lack of human biomonitoring data severely hinders the ability to interpret potential increases in exposure related to situations of environmental risk. We carried out a human biomonitoring study aimed to provide baseline information on background exposure of the Italian general population to this pollutant. For this purpose, we analysed 116 serum samples from groups of subjects of both sex from the general population residing in three Italian towns at different latitudes. Serum concentrations of beta-HCH resulted to be comprised between 1.64 and 300 ng/g fat, with a median value of 18.0 ng/g fat and a 90th percentile of 65.9 ng/g fat. The serum concentrations detected are in line with those detected in most Western European countries.

Although synaptic plasticity is widely regarded as the primary mechanism of memory [1], forms of nonsynaptic plasticity, such as increased somal or dendritic excitability or membrane potential depolarization, also have been implicated in learning in both vertebrate and invertebrate experimental systems [2-7]. Compared to synaptic plasticity, however, there is much less information available on the mechanisms of specific types of nonsynaptic plasticity involved in well-defined examples of behavioral memory. Recently, we have shown that learning-induced somal depolarization of an identified modulatory cell type (the cerebral giant cells, CGCs) of the snail Lymnaea stagnalis encodes information that enables the expression of long-term associative memory [8]. The Lymnaea CGCs therefore provide a highly suitable experimental system for investigating the ionic mechanisms of nonsynaptic plasticity that can be linked to behavioral learning. Based on a combined behavioral, electrophysiological, immunohistochemical, and computer simulation approach, here we show that an increase of a persistent sodium current of this neuron underlies its delayed and persistent depolarization after behavioral single-trial classical conditioning. Our findings provide new insights into how learning-induced membrane level changes are translated into a form of long-lasting neuronal plasticity already known to contribute to maintained adaptive modifications at the network and behavioral level [8].

Background: Morbid obesity, compromising cardiovascular and respiratory function, may increase the risk of anesthesia and was initially considered a contraindication to laparoscopy. The aim of this study was to investigate hemodynamic effects induced by pneumoperitoneum in superobese patients, assessed by arterial pulse contour method., Methods: We prospectively studied 10 obese patients (BMI 53 +/- 9 kg/m(2)), scheduled for laparoscopic gastric bypass. After anesthesia induction, patients were intubated and mechanically ventilated. A radial artery was cannulated to obtain hemodynamic data implemented by means of a new pulse contour analysis method-the pressure recording analytical method (PRAM). Data were recorded after anesthesia induction (Tbas), at peritoneal insufflation (T0), at 1, 3, 5, 10, 30, and 60 min after pneumoperitoneum induction (T1, T3, T5, T10, T15, T30, T60), at abdominal desufflation (Tdef) and 5 min after desufflation (T5def)., Results: MAP increased after pneumoperitoneum, returning to its baseline after deflation (79 +/- 7 Tbas, 81 +/- 6 mmHg T5def). HR remained unchanged. Systemic vascular resistance index (SVRI) increased after pneumoperitoneum induction and progressively returned to baseline (3,903 +/- 330 Tbas, 4,596 +/- 148 T1, and 3,786 +/- 202 dyn s m(2) cm(-5) T5def). Stroke volume index (SVI) and cardiac index (CI) increased after pneumoperitoneum and remained elevated. Stroke volume variation (SVV) decreased after insufflation remaining lower than the basal value (28 +/- 4% Tbas, 15 +/- 5% T5des). Cardiac Cycle Efficiency (CCE) transient decreased after insufflation and subsequently increased (0.029 +/- 0.146 Tbas, 0.008 +/- 0.178 T5def). Aortic dP/dt max increased after insufflation, returning to baseline only after desufflation (0.68 +/- 0.07 Tbas, 0.94 +/- 0.08 T30 and 0.86 +/- 0.06 mmHg s(-1) T5def)., Conclusion: As assessed by PRAM, this study showed no deterioration in hemodynamic indices or ventricular performance during laparoscopic gastric bypass.

In order to assess the correlation of pathological and radiological features of ductal carcinoma in situ (DCIS) of the breast and their association with surgical outcome, a consecutive series of 150 patients was retrospectively examined. Pathological slides from all patients were divided into three categories according to the pathological EPWG (European Pathologist Working Group) and DIN (Ductal Intraepithelial Neoplasia) classifications, which showed very good inter-correlation (r=0.99) (whole series). Mammographic images from 46 of these cases were blindly classified into five categories according to the level of radiological suspicion (R), morphology of calcifications (Ca) and preoperative results of needle biopsy (C/B) (limited series). No significant differences in the distribution of clinical and pathological variables were detected among whole and limited series. The lesions were grouped into two (low versus high) pathological (PRG), radiological (RRG and CaRG) and needle biopsy (C/BRG) risk groups. PRG was associated with both RRG (p=0.002) and CaRG (p=0000), but not with C/BRG. Correlations with surgical outcome were also explored, with lesions of high PRG being more likely to undergo re-excision for inadequate first wide local excision [odds ratio (OR)=2.1], mastectomy (OR=2.6) and nodal staging procedures (OR=3.8) in the whole series. Conversely, no significant correlation was found between PRG, RRG, CaRG and C/BRG with surgical outcome in the limited series. We suggest that pathological features of DCIS are associated with surgical outcome and may be predicted by mammography.

Forward genetic screens in model organisms have provided important insights into numerous aspects of development, physiology and pathology. With the availability of complete genome sequences and the introduction of RNA-mediated gene interference (RNAi), systematic reverse genetic screens are now also possible. Until now, such genome-wide RNAi screens have mostly been restricted to cultured cells and ubiquitous gene inactivation in Caenorhabditis elegans. This powerful approach has not yet been applied in a tissue-specific manner. Here we report the generation and validation of a genome-wide library of Drosophila melanogaster RNAi transgenes, enabling the conditional inactivation of gene function in specific tissues of the intact organism. Our RNAi transgenes consist of short gene fragments cloned as inverted repeats and expressed using the binary GAL4/UAS system. We generated 22,270 transgenic lines, covering 88% of the predicted protein-coding genes in the Drosophila genome. Molecular and phenotypic assays indicate that the majority of these transgenes are functional. Our transgenic RNAi library thus opens up the prospect of systematically analysing gene functions in any tissue and at any stage of the Drosophila lifespan.

Purpose: To prospectively compare clinical breast examination (CBE), mammography, ultrasonography (US), and contrast material-enhanced magnetic resonance (MR) imaging for screening women at genetic-familial high risk for breast cancer and report interim results, with pathologic findings as standard., Materials and Methods: Institutional review board of each center approved the research; informed written consent was obtained. CBE, mammography, US, and MR imaging were performed for yearly screening of BRCA1 or BRCA2 mutation carriers, first-degree relatives of BRCA1 or BRCA2 mutation carriers, or women enrolled because of a strong family history of breast or ovarian cancer (three or more events in first- or second-degree relatives in either maternal or paternal line; these included breast cancer in women younger than 60 years, ovarian cancer at any age, and male breast cancer at any age)., Results: Two hundred seventy-eight women (mean age, 46 years +/- 12 [standard deviation]) were enrolled. Breast cancer was found in 11 of 278 women at first round and seven of 99 at second round (14 invasive, four intraductal; eight were

We compared dynamic contrast-enhanced MRI (DCE-MRI) and sonography (US) for monitoring tumour size in 21 patients with breast cancer undergoing primary chemotherapy (PCT) followed by surgery. The correlation between DCE-MRI and US measurements of tumour size, defined as the product of the two major diameters, was 0.555 (P=0.009), 0.782 (P<0.001), and 0.793 (P<0.001) at baseline, and after two and four cycles of PCT, respectively. The median tumour size was significantly larger when measured by DCE-MRI than by US at baseline (1472 vs 900 mm(2), P<0.001) and after two cycles of PCT (600 vs 400 mm(2), P=0.009). After PCT, the median tumour size measured by the two techniques was similar (256 vs 289 mm(2) for DCE-MRI and US, respectively, P=0.859). The correlation with the histopathological major tumour diameter was 0.824 (P<0.001) and 0.705 (P<0.001) for post-treatment DCE-MRI and US, respectively. Measurements of the final major tumour diameter by DCE-MRI tended to be more precise, including cases achieving a pathological complete response. Randomized trials are warranted to establish the clinical impact of the initial discrepancy in tumour size estimates between DCE-MRI and US, and the trend towards a better definition of the final tumour size provided by DCE-MRI in this clinical setting.

Purpose: Dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) allows analysis of both tumor volume and contrast enhancement pattern using a single tool. We sought to investigate whether DCE-MRI could be used to predict histological response in patients undergoing primary chemotherapy (PCT) for breast cancer., Patients and Methods: Thirty patients with breast cancer, clinical diameter > 3 cm or stage III A/B, received anthracycline and taxane based PCT. DCE-MRI was performed at the baseline, after two cycles and after four cycles of PCT, before surgery. Histological response was assessed using a five-point scheme. Grade 4 (small cluster of dispersed residual cancer cells) and grade 5 (no residual viable cancer cell) were defined as a major histopathological response (MHR)., Results: Univariate analysis showed that a > 65% reduction in the tumor volume and a reduction in the early enhancement ratio (ECU) after two cycles of PCT were associated with a MHR. Multivariate analysis revealed that tumor volume reduction after two cycles of PCT was independently associated with a MHR (odds ratio [OR] 39.968, 95% confidence interval [CI] 3.438-464.962, p < 0.01). ECU reduction was still associated with a MHR (OR 2.50, 95% CI 0.263-23.775), but it did not retain statistical significance (p = 0.42). Combining tumor volume and ECU reduction after two cycles of PCT yielded a 93% diagnostic accuracy in identifying tumors achieving a pathological complete response (pCR) (histopathological grade 5)., Conclusions: DCE-MRI allows prediction of the effect of neoadjuvant chemotherapy in breast cancer. Although in our study tumor volume reduction after two cycles had the strongest predictive value, DCE-MRI has the potential to provide functional parameters that could be integrated to optimize neoadjuvant chemotherapy strategies.

Purpose: To evaluate if dynamic contrast enhanced magnetic resonance can predict the tumoural response in patients with breast cancer undergoing neoadjuvant chemotherapy., Materials and Methods: Twenty-six patients with biopsy-proven locally advanced breast cancer underwent taxane-based neoadjuvant chemotherapy followed by radical surgery. MRI evaluations were carried out at baseline, after two of the four planned cycles of chemotherapy, and before surgical treatment. MR images, obtained with 1.5 T scanner and using dedicated surface multichannel coil (GEMS), were acquired by 3D SPGR sequences on the coronal plane before and after intravenous administration of gadolinium chelate by an automatic injector. The percent reduction of the early contrast uptake between the baseline and the second MRI was then calculated for every lesion and correlated with the outcome of interest (pCR)., Results: At baseline, we identified 26 lesions and two patterns of MRI enhancement: homogeneous (group 1, 14 patients) and peripheral ring-like (group 2, 12 patients). At surgery, pathological complete response was observed in 4 patients (2 patients group 1, 2 patients group 2). In group 1, the 2 cases of pCR showed a gs;70% reduction in the early contrast uptake between baseline examination and after two cycles of chemotherapy, whereas values <70% were seen in 12 cases of partial response (p=0.02). In group 2, the 2 cases of pCR showed a gs; 40% reduction in the early contrast uptake whereas all of the 10 cases of partial response had <40% reduction (p=0.02)., Conclusions: In our series, the percent reduction in the early contrast uptake after two cycles of neoadjuvant chemotherapy was predictive of the achievement of a pathological complete response. However, we identified two distinct morphologic patterns associated with different cut-offs in the predictive value of the early contrast uptake reduction between baseline and intermediate examination.