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Collective user behavior in social media applications often drives several important online and offline phenomena linked to the spread of opinions and information. Several studies have focused on the analysis of such phenomena using networks to model user interactions, represented by edges. However, only a fraction of edges contribute to the actual investigation. Even worse, the often large number of non-relevant edges may obfuscate the salient interactions, blurring the underlying structures and user communities that capture the collective behavior patterns driving the target phenomenon. To solve this issue, researchers have proposed several network backbone extraction techniques to obtain a reduced and representative version of the network that better explains the phenomenon of interest. Each technique has its specific assumptions and procedure to extract the backbone. However, the literature lacks a clear methodology to highlight such assumptions, discuss how they affect the choice of a method and offer validation strategies in scenarios where no ground truth exists. In this work, we fill this gap by proposing a principled methodology for comparing and selecting the most appropriate backbone extraction method given a phenomenon of interest. We characterize ten state-of-the-art techniques in terms of their assumptions, requirements, and other aspects that one must consider to apply them in practice. We present four steps to apply, evaluate and select the best method(s) to a given target phenomenon. We validate our approach using two case studies with different requirements: online discussions on Instagram and coordinated behavior in WhatsApp groups. We show that each method can produce very different backbones, underlying that the choice of an adequate method is of utmost importance to reveal valuable knowledge about the particular phenomenon under investigation., Competing Interests: The authors have declared that no competing interests exist.

MicroRNAs (miRNAs) fine tune gene expression to regulate many aspects of nervous system physiology. Here, we show that miR-92a suppresses memory consolidation that occurs in the αβ and γ mushroom body neurons (MBns) of Drosophila , making miR-92a a memory suppressor miRNA. Bioinformatics analyses suggested that mRNAs encoding kinesin heavy chain 73 (KHC73), a protein that belongs to Kinesin-3 family of anterograde motor proteins, may be a functional target of miR-92a. Behavioral studies that employed expression of khc73 with and without its 3' untranslated region (UTR) containing miR-92a target sites, luciferase assays in HEK cells with reporters containing wild-type and mutant target sequences in the khc73 3'UTR, and immunohistochemistry experiments involving KHC73 expression with and without the wild-type khc73 3'UTR, all point to the conclusion that khc73 is a major target of miR-92a in its functional role as a miRNA memory suppressor gene., (Copyright © 2020 Guven-Ozkan et al.)

The COVID-19 pandemic led to the adoption of severe measures to counteract the spread of the infection. Social distancing and lockdown measures modified people's habits, while the Internet gained a major role in supporting remote working, e-teaching, online collaboration, gaming, video streaming, etc. All these sudden changes put unprecedented stress on the network. In this paper, we analyze the impact of the lockdown enforcement on the Politecnico di Torino campus network. Right after the school shutdown on the 25 th of February, PoliTO deployed its own in-house solution for virtual teaching. Ever since, the university provides about 600 virtual classes daily, serving more than 16 000 students per day. Here, we report a picture of how the pandemic changed PoliTO's network traffic. We first focus on the usage of remote working and collaboration platforms. Given the peculiarity of PoliTO online teaching solution that is hosted in-house, we drill down on the traffic, characterizing both the audience and the network footprint. Overall, we present a snapshot of the abrupt changes seen on campus traffic due to COVID-19, and testify how the Internet has proved robust to successfully cope with challenges while maintaining the university operations., Competing Interests: 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., (© 2020 Elsevier B.V. All rights reserved.)

Objectives: To assess, by means of biological monitoring, exposure to polycyclic aromatic hydrocarbons (PAHs) and to metallic elements in Italian Navy workers operating near the industrial area in Taranto, and thereby estimate the health risk., Methods: A total of 450 workers in the Italian Navy were examined; they had office type jobs, and 150 of them worked near the industrial area in Taranto (exposed group), 150 in Taranto but far from this area (internal control group) and 150 in Brindisi (external control group). The recruited workers were administered a questionnaire inquiring about current and previous working activities, personal medical history, lifestyle and dietary habits, and their residence location. Then they collected a urine sample for the determination of 1-hydroxypyrene, 2-naphthol, cotinine and the metallic elements As, Cd, Co, Cr, Mn, Ni, Pb, Cu, Zn and Hg. The latter were measured in 110 workers in each group. In addition, in some of the work sites of the three groups, environmental samplings were carried out to determine PAHs and the 10 metallic elements, also taking into account the wind direction., Results: Airborne benzo(a)pyrene concentrations at the different sampling sites ranged from 0.02 to 0.06 ng/m3 and naphthalene between <25 and 65.3 ng/m3, regardless of the wind direction. Among the metallic elements, As, Cd, Co, Cr, Hg, Pb, Cu and Zn were present at concentrations below or just above the limit of detection (LOD). Mn and Ni were slightly higher in the work sites of the exposed group. The urinary concentrations of 1-hydroxypyrene, 2-naphthol and the single metallic elements were not higher in the exposed workers group than in the other 2 groups. Smokers had significantly higher urinary 1-hydroxypyrene and 2-naphthol concentrations, whereas cigarette smoking did not condition a higher urinary elimination of metallic elements in the three groups with the exception of Cd and Pb. Moreover, residence location conditioned Mn, Hg and As urinary excretion, consumption of shellfish and/or crustaceans in the 72 hours before urine collection conditioned As elimination, and consumption of legumes in the 72 hours before urine collection conditioned Ni elimination., Conclusions: This research did not find a higher urinary excretion of 1-hydroxypyrene, 2-naphthol and As, Cd, Co, Cr, Mn, Ni, Pb, Cu, Zn and Hg in the exposed workers group as compared to the internal control group working far from the industrial area of Taranto, nor in the group working in another city far away from Taranto, Brindisi. Therefore, it indicated that workers in the Italian Navy operating near the industrial area in Taranto were not exposed to a greater risk attributable to exposure to PAHs and metallic elements than the two control groups.

This survey aimed to evaluate the applicability of the Nordic Occupational Skin Questionnaire (NOSQ) as a preliminary screening tool to investigate the presence of contact dermatological disorders in sea fishermen. The Italian version of the NOSQ was administered to 143 male fishermen working at an Apulia (Southern Italy) Fisheries, and 136 male workers who had never worked as sea fishers (controls). A significantly higher rate of frequency of transient itchy wheals on the hands, wrists, and forearms was recorded in the fishermen as compared to the controls (49.6% vs. 8.1%; p < 0.001), while there was no significant difference in the frequency of eczema (8.4% vs. 6.6%). In 46.1% of the fishermen, the onset of transient itchy wheals was associated with contact with specific agents and the most common causes were algae and aquatic plants (49.3%) and seabed sludge (25.3%). In conclusion, the administration of the NOSQ can be useful in preliminary screening for dermatitis in fishermen, although it could show a possible overestimation of the prevalence of transient itchy wheals., Competing Interests: The authors declare no conflict of interest.

Context: Urinary S-phenylmercapturic acid (SPMA) and benzene (U-Ben) are usually measured at the end of the work shift (ES), although their kinetic of elimination is not clearly known., Objective: To investigate SPMA and U-Ben elimination 16 h after the ES, in 93 coke production workers exposed to low benzene concentrations., Materials and Methods: Airborne benzene (A-Ben) was measured by passive samplings, while SPMA, U-Ben, methyl-tert-butyl ether (U-MTBE), cotinine (U-Cot) and creatinine were determined on urine samples collected at ES and before the beginning of the next work shift (next BS)., Results: Median A-Ben concentrations were 17.2 µg/m 3 in the personal and 34.7 µg/m 3 in the stationary samplings. SPMA was always detectable, whereas U-Ben was below the limit of quantification in 26.7% of the ES and 35.6% of the next BS samples, and U-MTBE in more than the 80.0% of the samples. At both the sampling times, SPMA and U-Ben showed a positive dependence on personal A-Ben, as well as on creatinine and U-Cot values., Discussion and Conclusion: SPMA and U-Ben at the next BS were dependent on the exposure to low benzene concentrations suffered in the previous work shift, prompting a reconsideration of the urine sampling time recommended by the American Conference Governmental Industrial Hygienists (ACGIH).

The contribution of biological and environmental monitoring to the risk assessment of occupational exposure to lead (Pb), arsenic (As), and antimony (Sb) was studied in 18 workers at a birdshot factory (Exposed) and in 18 control workers (Controls) by the determination of both airborne Pb (PbA) and airborne As (AsA) only in the exposed workers and blood Pb (PbB), erythrocytic zinc protoporphyrin (ZPP), urinary Sb (SbU), and the urinary As species in exposed workers and controls. PbA (12-42 μg/m 3 ) and AsA (1-4 μg/m 3 ) were strongly correlated (r = .95). PbB, ZPP, and the sum of As 3 +As 5 +MMA were significantly higher in the exposed workers. As 3 was higher than the limit of detection in 14 exposed workers and 1 control, As 5 only in 1 exposed worker, SbU in all the exposed workers and in 4 controls. Monitoring for more metallic elements reveals a wider spectrum of exposures than can be achieved by lead surveillance alone and is preferable for characterizing occupational risk wherever possible.

Changes in body temperature can profoundly affect survival. The dramatic longevity-enhancing effect of cold has long been known in organisms ranging from invertebrates to mammals, yet the underlying mechanisms have only recently begun to be uncovered. In the nematode Caenorhabditis elegans , this process is regulated by a thermosensitive membrane TRP channel and the DAF-16/FOXO transcription factor, but in more complex organisms the underpinnings of cold-induced longevity remain largely mysterious. We report that, in Drosophila melanogaster , variation in ambient temperature triggers metabolic changes in protein translation, mitochondrial protein synthesis, and posttranslational regulation of the translation repressor, 4E-BP (eukaryotic translation initiation factor 4E-binding protein). We show that 4E-BP determines Drosophila lifespan in the context of temperature changes, revealing a genetic mechanism for cold-induced longevity in this model organism. Our results suggest that the 4E-BP pathway, chiefly thought of as a nutrient sensor, may represent a master metabolic switch responding to diverse environmental factors., Competing Interests: The authors declare no conflict of interest.

The phenomenon of accidents at work was investigated among the resident physicians of the School of Medicine, Bari University, by a self-administered anonymous questionnaire probing personal details and inquiring about any accidents at work experienced during the training period, and by a comparison with the accidents reported to the Hospital Accidents Registry. At least 1 biological accident was reported by 18.2% of the 450 participants, this percentage being significantly higher in the surgical area (33.3%), where biological accidents were much more rarely reported to either the Residency School Director or the Accidents Registry. In conclusion, despite an overall reduction compared with the past, the frequency both of biological accidents and of underreporting is still high among resident physicians, particularly in the surgical area., (Copyright © 2016 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.)

The uptake of cytoplasmic calcium into mitochondria is critical for a variety of physiological processes, including calcium buffering, metabolism, and cell survival. Here, we demonstrate that inhibiting the mitochondrial calcium uniporter in the Drosophila mushroom body neurons (MBn)-a brain region critical for olfactory memory formation-causes memory impairment without altering the capacity to learn. Inhibiting uniporter activity only during pupation impaired adult memory, whereas the same inhibition during adulthood was without effect. The behavioral impairment was associated with structural defects in MBn, including a decrease in synaptic vesicles and an increased length in the axons of the αβ MBn. Our results reveal an in vivo developmental role for the mitochondrial uniporter complex in establishing the necessary structural and functional neuronal substrates for normal memory formation in the adult organism., (Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.)

The effects of occupational exposure to low concentrations of polychlorobiphenyls (PCBs) on the urinary excretion of corticosteroid hormones were evaluated, taking into account the influence of cigarette smoking. The study included 26 males working as electrical maintenance staff in a steel factory, previously exposed to a mixture of PCBs (exposed workers), and 30 male workers with no occupational exposure to PCBs (controls). Serum PCBs (33 congeners), urinary 17-hydroxycorticosteroids, 17-ketosteroids (KS) and pregnanes, and their respective glucuronidated and sulfonated compounds, were determined for each subject. PCBs were significantly higher in the exposed workers than controls, and were correlated with age. Both the urinary concentrations of the total 17-KS and pregnanes, and those of some single steroids and their glucuronidated compounds, were significantly lower in the exposed workers than controls, but higher in smokers than the non-smokers + ex-smokers. Two-way analysis of variance showed a negative association between serum PCBs and both total glucuronidated 17-KS and total and glucuronidated pregnanes, and a positive association between cigarette smoking and both total and glucuronidated 17-KS. PCBs seem to act as endocrine disruptors by reducing the urinary excretion of corticosteroid hormones, particularly of the glucuronidated fraction. Cigarette smoking could boost these effects of PCBs in smokers.

DNA damage and cellular repair capacity were studied in 18 male fuel tanker drivers and 13 male filling-station attendants exposed to low and very low concentrations of benzene, respectively, and compared to 20 males with no occupational exposure (controls). Exposure to airborne benzene was measured using passive personal samplers, and internal doses were assayed through the biomarkers t,t-muconic acid, S-phenylmercapturic acid and urinary benzene. DNA damage was evaluated using tail intensity (TI) determined by the comet assay in peripheral lymphocytes. Urinary 7-hydro-8-oxo-2'-deoxyguanosine (8-oxodG) was measured as a biomarker of oxidative damage. DNA repair kinetics were assessed using the comet assay in lymphocytes sampled 20 and 60 min post H2O2 exposure. Benzene exposure differed significantly between the drivers (median 246.3 µg/m(3)), attendants (median 13.8 µg/m(3)), and controls (median 4.1 µg/m(3)). There were no differences in TI and 8-oxodG among the three groups, or between smokers and non-smokers. DNA repair kinetics were similar among the drivers, attendants and controls, although the comet assay on H2 O2 -damaged lymphocytes after 60 min revealed significantly lower levels of TI only in drivers. The DNA repair process in smokers was similar to that observed in drivers. In conclusion, this study found no relationship between low levels of benzene exposure and DNA damage, although there was evidence that exposure interferes with DNA repair kinetics. The biological impact of this finding on the onset of genotoxic effects in exposed workers has still to be ascertained., (© 2015 Wiley Periodicals, Inc.)

Spinocerebellar ataxia type 28 (SCA28) is a neurodegenerative disease caused by mutations of the mitochondrial protease AFG3L2. The SCA28 mouse model, which is haploinsufficient for Afg3l2, exhibits a progressive decline in motor function and displays dark degeneration of Purkinje cells (PC-DCD) of mitochondrial origin. Here, we determined that mitochondria in cultured Afg3l2-deficient PCs ineffectively buffer evoked Ca²⁺ peaks, resulting in enhanced cytoplasmic Ca²⁺ concentrations, which subsequently triggers PC-DCD. This Ca²⁺-handling defect is the result of negative synergism between mitochondrial depolarization and altered organelle trafficking to PC dendrites in Afg3l2-mutant cells. In SCA28 mice, partial genetic silencing of the metabotropic glutamate receptor mGluR1 decreased Ca²⁺ influx in PCs and reversed the ataxic phenotype. Moreover, administration of the β-lactam antibiotic ceftriaxone, which promotes synaptic glutamate clearance, thereby reducing Ca²⁺ influx, improved ataxia-associated phenotypes in SCA28 mice when given either prior to or after symptom onset. Together, the results of this study indicate that ineffective mitochondrial Ca²⁺ handling in PCs underlies SCA28 pathogenesis and suggest that strategies that lower glutamate stimulation of PCs should be further explored as a potential treatment for SCA28 patients.

The frequency of chromosome aberrations (CA) and micronuclei (MN) was investigated in the peripheral lymphocytes of workers occupationally exposed to low or very low concentrations of benzene. The study included 43 exposed workers (all males), namely 19 fuel-tanker drivers and 24 filling-station attendants, and 31 male subjects with no occupational exposure to the toxicant (controls). Benzene exposure was verified by means of environmental monitoring with passive personal samplers (Radiello(®)), and through biological monitoring, i.e. by measurement of urinary trans,trans-muconic acid, S-phenylmercapturic acid and benzene. The frequency of CA and MN in peripheral lymphocytes was determined according to standard procedures. Exposure to benzene was found to be significantly higher for fuel-tanker drivers (median 246.6 μg/m(3)) than for filling-station attendants (median 19.9 μg/m(3)). Both groups had significantly higher exposure than controls (median 4.3 μg/m(3)). No increased frequency of CA and MN was observed in either fuel-tanker drivers or filling-station attendants compared with controls. In all subjects examined as a single group, the frequency of MN was significantly dependent on age. Only in the fuel-tanker drivers was the frequency of MN found to depend not only on age, but also on exposure to benzene. In conclusion, the frequency of MN, but not of CA, could be influenced by exposure to benzene concentrations of up to one order of magnitude lower than the threshold limit value (time-weighted average)., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Ca(2+) homeostasis in peroxisomes has been an unsolved problem for many years. Recently novel probes to monitor Ca(2+) levels in the lumen of peroxisomes in living cells of both animal and plant cells have been developed. Here we discuss the contrasting results obtained in mammalian cells with chemiluminecsent (aequorin) and fluorescent (cameleon) probes targeted to peroxisomes. We briefly discuss the different characteristics of these probes and the possible pitfalls of the two approaches. We conclude that the contrasting results obtained with the two probes may reflect a heterogeneity among peroxisomes in mammalian cells. We also discuss the results obtained in plant peroxisomes. In particular we demonstrate that Ca(2+) increases in the cytoplasm are mirrored by similar rises of Ca(2+) concentration the lumen of peroxisomes. The increases in peroxisome Ca(2+) level results in the activation of a catalase isoform, CAT3. Other functional roles of peroxisomal Ca(2+) changes in plant physiology are briefly discussed.

Objective: To study the urinary excretion of As, Cr, Mn, Co, Ni, Cu, Zn, Cd, Sn, Ba, Hg, Pb, Sb in workers at the Taranto integrated-cycle steel foundry and in subjects from the general population of Taranto, to assess the health risk posed by occupational exposure and environmental exposure, respectively, to these metals., Materials and Methods: The study included 49 steel foundry workers (exposed), working in the minerals and agglomerates pools, steel processing plants 1 and 2 and maritime plants, and 50 subjects belonging to the general population of Taranto resident at various distances from the factory (controls), randomly selected from the exposed subjects and controls enrolled in previous research conducted in 2005. A questionnaire was administered to all participants, enquiring into general characteristics, lifestyle, diet, and any medical conditions. Informed written consent to take part in the study was obtained from all subjects before enrolment. The results of environmental monitoring performed in 2005 in the workers' sectors, consisting of determining As, Cr, Mn, Ni, Cu, Zn, Cd and Pb in the respirable dust, revealed by both samplers applied in fixed positions and personal samplers, were considered. Urine samples were obtained from all participants on a Friday, to determine As and Cr by AAS and all the other metal elements by a multielement technique with ICP-MS. Urinary creatinine was also determined to make any necessary adjustments. All urine analyses were performed in 2005 within one month of urine collection., Results: In the respirable dust, As and Cd were always within the LOD, whereas Cr, Mn, Ni, Cu and Pb were 1-2 orders of magnitude below the respective TLV-TWA of the ACGIH. Mn was the only metal element that presented significantly higher urinary concentrations in exposed subjects as compared to controls, although the values in both groups were in any case within the Italian reference range. Co, Cu, Zn, Sn and Sb showed significantly higher urinary concentrations in controls than in the exposed subjects, while there were no differences in As, Cr, Cd, Ba, Hg and Pb between the two groups. Ni was within the LOD in the 60% of the subjects in the two groups. Multiple stepwise regression showed a dependence of Cr on age, of As on the number of days since the last meal of mollusks and crustaceans, of Zn on the consumption of mollusks, and of Cd and Pb on smoking expressed as the number of pack/year., Discussion: The research did not reveal higher urinary excretion of the metal elements in the steelworkers than in the subjects from the general population of Taranto resident at different distances from the factory, except for Mn, that was anyway within the reference range. This allows us to consider that there is no relevant health risk posed by occupational exposure to metal elements in the steelworkers, nor by environmental exposure to the same elements in the residents of the city of Taranto. In particular, no pollution of the living environment by metal elements of industrial origin was demonstrated, whereas other non occupational factors seem to condition the intake of these metal elements.

Objective: To assess the critical issues concerning the use of urinary inorganic arsenic (iAs), including As3, As5, monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA), as biomarker of internal dose in order to monitor environmental and occupational exposure to inorganic As, considering the influence of diet and drinking water on excretion of iAs., Methods: The design protocol stipulated collection of weekly urine samples from 6 male subjects for 5 consecutive months. In all the urine samples, iAs was determined by Hydride Generation-Atomic Absorption Spectrophotometry (HG-AAS). In the subjects with iAs higher than 35 microg/L, Biological Exposure Index (BEI) proposed by the American Conference of Governmental Industrial Hygienists (ACGIH), urinary arsenic speciation was performed by HPLC-ICP-MS. Exposure to airborne As was evaluated monthly using personal environmental samplers worn for 8 hours. Throughout the study, the participants filled out a daily food diary, also detailing types of water drunk., Result: Exposure to airborne As was invariably below the limit of detection, equal to 1 ng/m3. A total of 77 urine samples were collected. iAs was always detectable and was higher in 7 urine samples, obtained from 5 of the 6 subjects examined, than the BEI. Among foods with a high As content, the intake of seafood and fish within 72 hours before providing the sample seems to be the principal source of the iAs concentrations, while the intake of rice or drinking water showed no influence on this biological marker. Instead, drinking wine within 24 hours before urine sample collection can cause a significant increase in the excretion of iAs., Conclusions: In populations that eat large amounts of fish and seafood, the use of iAs to monitor occupational and environmental exposure to inorganic As seems to present some problems, and urinary As speciation may be essential at least in cases with As measurements above the biological limit values. In any case, a diet sheet reporting all foods eaten within 3 days of urine collection seems to be an indispensable tool to ensure a correct interpretation of the results.

Mitochondrial ability of shaping Ca(2+) signals has been demonstrated in a large number of cell types, but it is still debated in heart cells. Here, we take advantage of the molecular identification of the mitochondrial Ca(2+) uniporter (MCU) and of unique targeted Ca(2+) probes to directly address this issue. We demonstrate that, during spontaneous Ca(2+) pacing, Ca(2+) peaks on the outer mitochondrial membrane (OMM) are much greater than in the cytoplasm because of a large number of Ca(2+) hot spots generated on the OMM surface. Cytoplasmic Ca(2+) peaks are reduced or enhanced by MCU overexpression and siRNA silencing, respectively; the opposite occurs within the mitochondrial matrix. Accordingly, the extent of contraction is reduced by overexpression of MCU and augmented by its down-regulation. Modulation of MCU levels does not affect the ATP content of the cardiomyocytes. Thus, in neonatal cardiac myocytes, mitochondria significantly contribute to buffering the amplitude of systolic Ca(2+) rises.

Here we describe use of a mitochondrial targeted Cameleon to produce stably transformed Arabidopsis plants that enable analyses of mitochondrial Ca²⁺ dynamics in planta and allow monitoring of the intra-mitochondrial Ca²⁺ concentration in response to physiological or environmental stimuli. Transgenic plants co-expressing nuclear and mitochondrial targeted Cameleons were also generated and analyzed. Here we show that mitochondrial Ca²⁺ accumulation is strictly related to the intensity of the cytoplasmic Ca²⁺ increase, demonstrating a tight association between mitochondrial and cytoplasmic Ca²⁺ dynamics. However, under all experimental conditions, mitochondrial Ca²⁺ dynamics were substantially different from those monitored in the cytoplasm, demonstrating that mitochondria do not passively sense cytosolic Ca²⁺, but actively modulate the intra-mitochondrial level of the cation. In particular, our analyses show that the kinetics of Ca²⁺ release from mitochondria are much slower than in the cytoplasm and nucleus. The mechanisms and functional implications of these differences are discussed., (© 2012 The Authors. The Plant Journal © 2012 Blackwell Publishing Ltd.)

Mitochondria from every tissue are quite similar in their capability to accumulate Ca²⁺ in a process that depends on the electrical potential across the inner membrane; it is catalyzed by a gated channel (named mitochondrial Ca²⁺ uniporter), the molecular identity of which has only recently been unraveled. The release of accumulated Ca²⁺ in mitochondria from different tissues is, on the contrary, quite variable, both in terms of speed and mechanism: a Na⁺-dependent efflux in excitable cells (catalyzed by NCLX) and a H⁺/Ca²⁺ exchanger in other cells. The efficacy of mitochondrial Ca²⁺ uptake in living cells is strictly dependent on the topological arrangement of the organelles with respect to the source of Ca²⁺ flowing into the cytoplasm, i.e., plasma membrane or intracellular channels. In turn, the structural and functional relationships between mitochondria and other cellular membranes are dictated by the specific architecture of different cells. Mitochondria not only modulate the amplitude and the kinetics of local and bulk cytoplasmic Ca²⁺ changes but also depend on the Ca²⁺ signal for their own functionality, in particular for their capacity to produce ATP. In this review, we summarize the processes involved in mitochondrial Ca²⁺ handling and its integration in cell physiology, highlighting the main common characteristics as well as key differences, in different tissues.

To study the effect of eating foods with a high arsenic (As) content on the intra and inter-individual variability of urinary concentrations of the As species, daily urine samples were collected for 10 consecutive days from 12 healthy male subjects. A daily food diary was kept throughout the study period. Personal exposure to airborne As was measured once during the study. As(3), As(5), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA) and arsenobetaine were determined in all urine samples by inductive coupled plasma mass spectrometry, and the sum of As(3)+As(5)+MMA+DMA (iAs) by hydride generation-atomic absorption spectrophotometry. Exposure to airborne As was below the limit of detection in all samplings. As(3) was found in only 19.2% and As(5) in only 3.3% of the urine samples, whereas high urinary concentrations of arsenobetaine were observed. With the exception of arsenobetaine, expressed as a percentage, a significant inter-individual variability was observed for all species of As, for iAs and for the MMA/DMA ratio (p<0.001). Instead, the intra-individual variability was significant only for the MMA/DMA ratio (p<0.001). Among foods with a high As content, only a heavy consumption of seafood was shown to influence inter-individual variability of DMA%, arsenobetaine expressed as μg g(-1) creatinine and iAs. In conclusion, even in populations with a high intake of organic As through foods, the finding of a significant inter-individual but no significant intra-individual variability of urinary species confirms the usefulness of urinary As speciation for biological monitoring of exposure to As., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The urinary excretion of t,t-muconic acid (t,t-MA), S-phenylmercapturic acid (SPMA) and urinary benzene and the influence of a smoking habit and of exposure to urban traffic on the urinary excretion of these biomarkers were investigated in 137 male adults from the general population. All subjects were not occupationally exposed to benzene and resident in two cities in Puglia (Southern-Italy). Environmental exposure to benzene was measured using passive personal samplers. The biomarkers t,t-MA, SPMA and urinary benzene were determined in urine samples collected from each subject at the end of the environmental sampling. The percentage of cases above the limit of detection was higher for SPMA and urinary benzene in smokers than in non-smokers, and for airborne benzene and urinary benzene in subjects exposed to urban traffic. Airborne benzene was correlated with the time spent in urban traffic during the environmental sampling. Among the biomarkers, urinary benzene was found to be correlated with airborne benzene only in non-smokers, and with the time spent in urban traffic, both in smokers and non-smokers considered together, and in non-smokers only. Finally, multiple regression analysis showed that the urinary excretion of all the biomarkers was dependent on the number of cigarettes smoked per day and, for urinary benzene, also on the time spent in urban traffic. In conclusion, urinary benzene seems to be a more valid biomarker than t,t-MA and SPMA to assess environmental exposure to extremely low concentrations of benzene. Cigarette smoking prevailed over traffic exhaust fumes in determining the internal dose of benzene.

Mitochondrial Ca(2+) uptake and release play a fundamental role in the control of different physiological processes, such as cytoplasmic Ca(2+) signalling, ATP production and hormone metabolism, while dysregulation of mitochondrial Ca(2+) handling triggers the cascade of events that lead to cell death. The basic mechanisms of mitochondrial Ca(2+) homeostasis have been firmly established for decades, but the molecular identities of the channels and transporters responsible for Ca(2+) uptake and release have remained mysterious until very recently. Here, we briefly review the main findings that have led to our present understanding of mitochondrial Ca(2+) homeostasis and its integration in cell physiology. We will then discuss the recent work that has unravelled the biochemical identity of three key molecules: NCLX, the mitochondrial Na(+)/Ca(2+) antiporter, MCU, the pore-forming subunit of the mitochondrial Ca(2+) uptake channel, and MICU1, one of its regulatory subunits.

Aim: To study the validity of urinary benzene as a biomarker of low and very low exposure to this toxicant, as compared with t,t-muconic acid (t,t-MA) and S-phenylmercapturic acid (SPMA), also taking into account the influence of cigarette smoking and co-exposure to toluene on the urinary excretion of benzene., Materials and Methods: The results obtained in two different studies were compared: in the first, workers occupationally exposed to low concentrations of benzene (18 fuel tanker drivers and 23 filling station attendants) were compared with 31 controls and in the second, workers exposed to very low concentrations of benzene (the same 23 filling station attendants) were compared with the 31 controls. Exposure to airborne benzene and toluene was monitored with passive personal samplers (Radiello). Then the urine collected at the end of the work shift was analyzed for t,t-MA, SPMA and urinary benzene. All participants also filled out a questionnaire about their lifestyle habits., Results: There were no differences among the three groups in terms of age and smoking habit. Occupational exposure to benzene and toluene and the urinary concentrations of t,t-MA, SPMA and urinary benzene were higher in the fuel tanker drivers than the filling station attendants and higher in the latter than in the controls. Cigarette smoking was found to be associated with urinary excretion of t,t-MA, SPMA and urinary benzene at both low and very low exposure to benzene. The biomarkers t,t-MA, SPMA and urinary benzene were almost always correlated, for both low and very low exposure to benzene. Notably, for low exposure to benzene a dependency relation was found with the levels of t,t-MA, SPMA and urinary benzene on both cigarette smoking and airborne benzene, whereas for very low exposure to benzene there was a dependency relation of SPMA on cigarette smoking and airborne benzene, of urinary benzene only on cigarette smoking and of t,t-MA on none of the variables considered., Conclusions: For occupational exposure to low concentrations of benzene, urinary benzene and SPMA showed a comparable validity, while for exposure to very low concentrations of this toxicant the validity of SPMA was confirmed while urinary benzene was found to be less useful. Cigarette smoking was the main factor conditioning the excretion of all the biomarkers of benzene in conditions of both low and very low exposure to the toxicant, so for the analysis of occupational exposure to benzene it is best to recommend abstention from smoking at least in the hours immediately before urine collection.

Oxidative stress is a major challenge for all cells living in an oxygen-based world. Among reactive oxygen species, H2O2, is a well known toxic molecule and, nowadays, considered a specific component of several signalling pathways. In order to gain insight into the roles played by H2O2 in plant cells, it is necessary to have a reliable, specific and non-invasive methodology for its in vivo detection. Hence, the genetically encoded H2O2 sensor HyPer was expressed in plant cells in different subcellular compartments such as cytoplasm and peroxisomes. Moreover, with the use of the new green fluorescent protein (GFP)-based Cameleon Ca2+ indicator, D3cpv-KVK-SKL, targeted to peroxisomes, we demonstrated that the induction of cytoplasmic Ca2+ increase is followed by Ca2+ rise in the peroxisomal lumen. The analyses of HyPer fluorescence ratios were performed in leaf peroxisomes of tobacco and pre- and post-bolting Arabidopsis plants. These analyses allowed us to demonstrate that an intraperoxisomal Ca2+ rise in vivo stimulates catalase activity, increasing peroxisomal H2O2 scavenging efficiency.

Calcium handling by mitochondria is a key feature in cell life. It is involved in energy production for cell activity, in buffering and shaping cytosolic calcium rises and also in determining cell fate by triggering or preventing apoptosis. Both mitochondria and the mechanisms involved in the control of calcium homeostasis have been extensively studied, but they still provide researchers with long-standing or even new challenges. Technical improvements in the tools employed for the investigation of calcium dynamics have been-and are still-opening new perspectives in this field, and more prominently for mitochondria. In this review we present a state-of-the-art toolkit for calcium measurements, with major emphasis on the advantages of genetically encoded indicators. These indicators can be efficiently and selectively targeted to specific cellular sub-compartments, allowing previously unavailable high-definition calcium dynamic studies. We also summarize the main features of cellular and, in more detail, mitochondrial calcium handling, especially focusing on the latest breakthroughs in the field, such as the recent direct characterization of the calcium microdomains that occur on the mitochondrial surface upon cellular stimulation. Additionally, we provide a major example of the key role played by calcium in patho-physiology by briefly describing the extensively reported-albeit highly controversial-alterations of calcium homeostasis in Alzheimer's disease, casting lights on the possible alterations in mitochondrial calcium handling in this pathology., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Although it is widely accepted that mitochondria in living cells can efficiently uptake Ca(2+) during stimulation because of their vicinity to microdomains of high [Ca(2+)], the direct proof of Ca(2+) hot spots' existence is still lacking. Thanks to a GFP-based Ca(2+) probe localized on the cytosolic surface of the outer mitochondrial membrane, we demonstrate that, upon Ca(2+) mobilization, the [Ca(2+)] in small regions of the mitochondrial surface reaches levels 5- to 10-fold higher than in the bulk cytosol. We also show that the [Ca(2+)] to which mitochondria are exposed during capacitative Ca(2+) influx is similar between near plasma membrane mitochondria and organelles deeply located in the cytoplasm, whereas it is 2- to 3-fold higher in subplasma membrane mitochondria upon activation of voltage-gated Ca(2+) channels. These results demonstrate that mitochondria are exposed to Ca(2+) hot spots close to the ER but are excluded from the regions where capacitative Ca(2+) influx occurs., (Copyright 2010 Elsevier Inc. All rights reserved.)

Objectives: This study analyzes the validity of new, more sensitive and specific urinary biomarkers of internal dose, namely, urinary benzene for benzene and urinary toluene and S-benzylmercapturic acid (SBMA) for toluene, to assess their efficacy when compared to traditional biomarkers for biological monitoring of occupational exposure to low concentrations of these two toxic substances., Methods: Assessment was made of 41 workers occupationally exposed to benzene and toluene, 18 fuel tanker drivers and 23 filling-station attendants, as well as 31 subjects with no occupational exposure to these toxic substances (controls). Exposure to airborne benzene and toluene was measured using passive Radiello personal samplers worn throughout the work shift. In urine samples collected from all subjects at the end of the workday, both the traditional and the new internal dose biomarkers of benzene and toluene were assessed, as well as creatinine so as to apply suitable adjustments., Results: Occupational exposure to benzene and toluene resulted significantly higher in the fuel tanker drivers than the filling-station attendants, and higher in the latter than in controls. Significantly higher concentrations of t,t-muconic acid (t,t-MA), S-phenylmercapturic acid (SPMA), urinary benzene, SBMA and urinary toluene were found in the drivers than the filling-station attendants or the controls. Instead, urinary phenol and hippuric acid were not different in the three groups. In the entire sample, airborne benzene and toluene values were significantly correlated, as were the respective urinary biomarkers, showing coefficients ranging from 0.36 to 0.98. Subdividing the subjects by smoking habit, higher coefficients were evident in non-smokers than in smokers; at multiple regression analysis t,t-MA, SPMA and urinary benzene and toluene were dependent on the number of cigarettes smoked daily and on airborne benzene and toluene, respectively. Instead, SBMA was dependent only on airborne toluene., Conclusions: Our research confirmed the validity of t,t-MA and SPMA for use in the biological monitoring of exposure to low concentrations of benzene. Urinary benzene showed comparable validity to SPMA; both parameters are affected by smoking cigarettes in the hours before urine collection, so it is best to ask subjects to refrain from smoking for 2 h before urine collection. Urinary toluene was found to be a more specific biomarker than SBMA.

Indoor and outdoor formaldehyde and acetaldehyde levels were assessed to characterize pollution in dwellings in the city and the Province of Bari, also taking into account seasonal variability, and to investigate health effects of aldehyde exposure on the general population. In 2007, passive environmental monitoring was performed, for 24 hours, in the kitchen of 59 dwellings, as well as outdoors for 27 of them. A questionnaire probing personal and home characteristics was administered to all 182 subjects habitually resident in the homes. During the period January-June 2008, formaldehyde and acetaldehyde levels were monitored monthly in 20 of the investigated dwellings inhabited only by non smokers. Indoor formaldehyde and acetaldehyde concentrations were significantly higher (16.0 +/- 8.0 and 10.7 +/- 8.8 microg m(-3)) than outdoor concentrations (4.4 +/- 1.7 and 3.4 +/- 2.0 microg m(-3)), showing a correlation between indoor levels of the two aldehydes (r = 0.41; p = 0.001). In dwellings inhabited only by non smokers, formaldehyde concentrations were higher in the presence of furniture bought new or restored less than one year before (p = 0.03). Formaldehyde and acetaldehyde levels were significantly higher in winter months than in spring-summer months (F = 2.86, p = 0.02; F = 5.39, p < 0.001) and seemed to be influenced by the time that kitchen windows were kept open. As regards the effects on human health, a low prevalence of allergic disease and no association between any irritant or allergic complaints and indoor levels of the two aldehydes was observed. In conclusion, the results showed low indoor and outdoor concentrations of formaldehyde and acetaldehyde, not generally posing a risk for human health.

In order to characterize the different sources of exposure to arsenic (As), urinary excretion of total As, the sum of inorganic As+MMA+DMA determined by the hydride generation-atomic absorption spectrophotometry technique, and the species As3, As5, monomethylarsonic acid (MMA), dimethylarsinic acid (DMA) and arsenobetaine were determined in 49 workers at a steel foundry, with presumed occupational exposure to As, and 50 subjects from the general population, all males. No evidence of occupational exposure to As resulted from environmental monitoring performed in the foundry, although the analysis of minerals used as raw materials showed the presence of As, particularly in fossils and fine ores. The urinary concentrations of As3, MMA, DMA, the sum of inorganic As+MMA+DMA and total As were not different in the two groups, while arsenobetaine appeared significantly higher in the controls than in the workers. The different species of urinary As were all significantly correlated. Urinary excretion of As3 was associated with the consumption of mineral water and with residence in an industrial zone, while MMA, DMA, arsenobetaine, the sum of inorganic As+MMA+DMA and total As urinary excretion were associated with the consumption of crustaceans and/or shellfish 3 days or less before urine collection. Multiple regression analysis confirmed these results. In conclusion, in populations with a high consumption of seafood, living in areas characterized by coastal/marine As pollution, only speciation of As can identify a prevalent role of environmental sources, like the consumption of seafood contaminated by As, in determining urinary As excretion, and exclude an occupational origin of the exposure.

We here describe the generation of novel, green fluorescent protein-based Ca(2+) indicators targeted to the peroxisome lumen. We show that (i) the Ca(2+) concentration of peroxisomes in living cells at rest is similar to that of the cytosol; (ii) increases in cytosolic Ca(2+) concentration (elicited by either Ca(2+) mobilization from stores or Ca(2+) influx through plasma membrane Ca(2+) channels) are followed by a slow rise in intraperoxisomal [Ca(2+)]; (iii) Ca(2+) influx into peroxisomes is driven neither by an ATP-dependent pump nor by membrane potential nor by a H(+)(Na(+)) gradient. The peroxisomal membrane appears to play a low pass filter role, preventing the organelle from taking up shortlasting cytosolic Ca(2+) transients but allowing equilibration of the peroxisomal luminal [Ca(2+)] with that of the cytosol during prolonged Ca(2+) increases. Thus, peroxisomes appear to be an additional cytosolic Ca(2+) buffer, but their influx and efflux mechanisms are unlike those of any other cellular organelle.

Creatinine adjustment has been used to remove the influence of the effect of urine dilution on exposure biomarkers measured in spot samples. This research aimed to determine the reliability of creatinine adjustment for urinary chromium and arsenic in subjects from general population considering interferences able to influence creatinine excretion. 444 male subjects were examined and each participant was administered an anamnestic questionnaire. Chromium and arsenic were determined on second morning void urine samples by atomic absorption spectrophotometry and creatinine by Jaffè method. Urinary creatinine showed a significant negative decrease with age increasing (p < 0.001). Chromium concentrations resulted higher in subjects with renal disease, particularly after creatinine adjustment (p = 0.014). Age was negatively correlated with urinary creatinine (rho = -0.23; p < 0.001), and positively with chromium (rho = 0.13; p = 0.007) and arsenic (rho = 0.17; p = 0.004), only after creatinine adjustment. A positive correlation was also found between unadjusted chromium and arsenic (rho = 0.10; p = 0.039) and between urinary creatinine and both unadjusted chromium (rho = 0.32; p < 0.001) and arsenic (rho = 0.18; p < 0.001). An high coefficient of correlation was observed between unadjusted chromium (rho = 0.88; p < 0.001) and arsenic (rho = 0.90; p < 0.001) and the respective adjusted values. Multiple regression showed a dependence of urinary creatinine and adjusted chromium and arsenic concentrations on age. In conclusion, although chromium and arsenic seem to have a renal kinetics of excretion comparable to the creatinine one, the influence of age on creatinine elimination suggests that the expression of urinary values of these metallic elements as microg/L is more reliable.

The influence of cigarette smoking on concentrations of urinary benzene, a sensitive and specific biomarker proposed for biological monitoring of exposure to very low doses of benzene, was investigated in 24 filling-station attendants and 31 workers non occupationally exposed to benzene. Environmental monitoring was performed by personal passive samplers "Radiello", and a spot urine sample was collected at the end of the work shift, from all subjects, for the determination of urinary benzene. Exposure to benzene resulted significantly higher in filling-station attendants (mean 23.3 +/- 17.0 microg/m3; range 4.5-66.3 microg/m3) than in controls (mean 4.6 +/- 2.6 microg/m3; range < 3-11.5 microg/m3), while concentrations of urinary benzene did not show any significant difference between the two groups. Considering all subjects as a single group, it was observed that urinary benzene concentrations were positively correlated with the number of cigarettes smoked during the sampling time (rho = 0.38; p = 0.047) and with airborne benzene levels (rho = 0.32; p = 0.019), and negatively correlated with the time elapsed between the last smoked cigarette and urine collecting (rho = -0.40; p = 0.045). Multiple regression analysis confirmed the influence of cigarette smoking on urinary benzene concentrations. In conclusion, our study showed the validity of urinary benzene as a biomarker for biological monitoring of exposure to very low doses of benzene, although cigarette smoking determined a prevalent etiological role at the low environmental benzene concentrations observed.

1,3-Butadiene (BD), a probable carcinogen to humans, has been shown to have an ill-defined genotoxicity in occupationally exposed workers. In the present study, the influence of exposure to very low doses of BD and to cigarette smoking was investigated on some cytogenetic endpoints, namely, sister chromatid exchanges (SCE), chromosomal aberrations (CA) and cells with a high frequency of SCE (HFC), in peripheral blood lymphocytes. Twenty-seven male workers employed in a petrochemical plant and 26 matched controls were included in the study. As regards the airborne BD values, there was a significant difference between exposed (median BD value 1.5, min-max 0.2-69.0 microg/m3) and non-exposed workers (median BD value 0.4, min-max <0.1-3.8 microg/m3). Genotoxic biomarkers were not able to distinguish between the two groups. The frequency of SCE was higher in smokers than in non-smokers (p=0.001), with a positive correlation between the number of cigarettes smoked per day and both SCE (r=0.4; p=0.004) and HFC frequency (r=0.3; p=0.04). Multiple regression analysis confirmed the influence of cigarette smoking on the level of SCE and HFC, while these parameters were not affected by personal exposure to BD. Overall, the biomarkers of genotoxic effect investigated in our study were not able to discriminate between workers with a very low exposure to BD and controls, while it was possible to distinguish between smokers and non-smokers on the basis of SCE.

Introduction: Urinary inorganic arsenic is an expression of occupational exposure to the metal, provided that there is no history of ingestion of foods containing high concentrations of inorganic and/or organic arsenic. The present study was conducted to assess the contribution of professional and environmental exposure to inorganic arsenic on urinary excretion of the metal., Materials and Methods: We examined 195 workers at a steelfoundry in Taranto, exposed to very low concentrations of inorganic arsenic and two control groups consisting of 105 subjects resident near the factory and 144 subjects resident approximately 20 Km away. All participants were administered a questionnaire enquiring about general characteristics, lifestyle, occupational and extra occupational exposure to arsenic. Urinary arsenic was determined by atomic-absorption spectrophotometry., Results: Exposed and non exposed subjects were similar as regards general characteristics and lifestyle. The environmental concentration of arsenic for the foundryworkers was invariably lower than 0.1 microg/m3. Urinary excretion of arsenic was higher in the subjects in all three groups, examined singly and together, if they had eaten crustaceans and/or shellfish in the three days before urine collection. There was a significant positive correlation with the consumption of shellfish and a significant negative correlation with the number of days since the last crustacean/shellfish meal. Multiple regression analysis showed a dependence of urinary elimination of arsenic on the days since the last crustacean/shellfish meal., Discussion: The absence of occupational exposure to arsenic allowed us to attribute the higher urinary elimination of arsenic to ingestion of crustaceans and/or shellfish in the three days before collection of the urine, both in subjects exposed to inorganic arsenic and in the two groups belonging to the general population. Our results support the hypothesis that inorganic arsenic, determined by atomic-absorption spectrophotometry, may derive from the catabolism of organic arsenic compounds contained in crustaceans and/or shellfish included in the diet.

The aim of the research was to assess the contribution of dental amalgams and other non-occupational factors of exposure to inorganic mercury (diet, etc.) to the quantity of mercury excreted with urine in workers exposed to low level concentrations of inorganic mercury. Two groups of workers (Groups I and II) were studied who were exposed to low and different environmental concentrations of inorganic mercury. These two groups were compared with a group of subjects not occupationally exposed to mercury in the same geographical area (Group III). All subjects were administered a questionnaire concerning personal data, lifestyle, recent removal and/or insertion of dental amalgam fillings, presence of nasal obstruction or bruxism and consumption of fish. The number of amalgam-filled teeth was established for each subject. Mean environmental exposure to inorganic mercury was 0.0087 mg/m3 for Group I and 0.0030 mg/m3 for Group II. Urinary excretion in the 3 groups was 4.2 +/- 2.8 micrograms/l for Group I, 3.0 +/- 2.1 micrograms/l for Group II and 1.6 +/- 1.2 micrograms/l for Group III. The results showed that of the factors of exposure to inorganic mercury, only occupational exposure (T = 9.18; p = 0.000) and the number of amalgam-filled teeth (T = 2.03; p = 0.043) were able to influence significantly urinary excretion of mercury; the sources of non-occupational exposure did not appear to play any role. The contribution of each amalgam filling to urinary mercury excretion was calculated to be 0.08 microgram/l. Occupational exposure therefore, even at low level doses, is still the main cause of urinary mercury excretion in workers exposed to inorganic mercury; of the non-occupational exposure factors, a significant role is played by amalgam dental fillings, whose contribution needs to be taken into consideration in order to make a correct interpretation of the results of biological monitoring of exposed workers.

The purpose of this study was to assess the role of dental amalgams and diet upon urinary mercury (U-Hg) excretion. 98 subjects (50 men and 48 women) not exposed to inorganic mercury, for either occupational or environmental reasons, and living in coastal and inland districts of Apulia (Southern Italy) were considered. All the subjects were administered a questionnaire with questions concerning life style, medical history, and occupational activity. Dental amalgams were evaluated with respect to their number and their surface areas. Urinary mercury was measured by the cold vapour atomic absorption technique. Expressed in terms of arithmetic mean, U-Hg excretion was found to amount to 1.03 micrograms/g creatinine (5th and 95th percentile: 0.31 and 2.40; range 0.30-3.25). Multiple linear regression analysis showed that, of the several tested independent variables (dental amalgams, age, body mass index, consumption of tuna, bass, swordfish, etc.), only the number of amalgam fillings (T = 5.25; p = 0.025) and the number of restored surfaces (T = 2.33; p = 0.020) were found liable to affect urinary mercury excretion in a significant manner. In conclusion, the results of this study confirm the primary role of amalgam fillings in affecting urinary mercury excretion in those subjects who are not occupationally exposed to inorganic mercury, The resulting urinary mercury levels can no doubt be taken as the reference values for the population of Apulia.