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This study aimed at surveying lower secondary schools in southern Italy, in a highly polluted area. A community close to an industrial area and three villages in rural areas was investigated. Indoor temperature, relative humidity (RH), gaseous pollutants (CO 2 and NO 2 ), selected biological pollutants in indoor dust, and the indoor/outdoor mass concentration and elemental composition of PM 2.5 were ascertained. Temperature and RH were within, or close to, the comfort range, while CO 2 frequently exceeded the threshold of 1000ppm, indicating inadequate air exchange rate. In all the classrooms, median NO 2 levels were above the WHO threshold value. Dermatophagoides p. allergen concentration was below the sensitizing threshold, while high endotoxin levels were detected in the classrooms, suggesting schools may produce significant risks of endotoxin exposure. Concentration and solubility of PM 2.5 elements were used to identify the sources of indoor particles. Indoor concentration of most elements was higher than outdoors. Resuspension was responsible for the indoor increase in soil components. For elements from industrial emission (Cd, Co, Ni, Pb, Sb, Tl, V), the indoor concentration depended on penetration from the outside. For these elements, differences in rural vs industrial concentrations were found, suggesting industrial sources may influence indoor air quality nearby schools.

Physical fitness values by the 6 min Walking Test (6MWT) at baseline (BL) and week-12 (W12) in the walk group divided by gender. Values as expressed as median (Q1-Q3). W12 values were compared to BL values by the Wilcoxon matched-pairs signed rank test. a. At BL, women had higher %HRmean than men (p = 0.021, Mann–Whitney test). HR, heart rate; Δ[La−], difference in lactate blood concentration between before and after 6MWT; Δ RPE, difference in Rate of Perceived Exertion between before and after 6MWT. (DOCX 48 kb)

Values of inflammatory markers at baseline (BL) and week-12 (W12) in the walk group divided by gender. Values as expressed as median (Q1-Q3). W12 values were compared to BL values by the Wilcoxon matched-pairs signed rank test. b. A subset of women (n = 3) and men (n = 7) were tested for cell activation markers. HsCRP, high sensitivity C-reactive protein; IL-6, interleukin-6; IL-8, interleukin-18; sCD14, soluble CD14. (DOCX 51 kb)

1-Repetition Maximum Test performance at baseline (BL) and week-12 (W12). Values are expressed as median (Q1-Q3). W12 values were compared to BL values by the Wilcoxon matched-pairs signed rank test. HR, heart rate; Î [Laâ ], difference in lactate blood concentration between before and after 6MWT; Î RPE, difference in Rate of Perceived Exertion between before and after 6MWT. (DOCX 39 kb)

Body composition and laboratory values at baseline (BL) and week-12 (W12) in the walk group divided by gender. Values as expressed as median (Q1-Q3). W12 values were compared to BL values by the Wilcoxon matched-pairs signed rank test. a. At BL, women had higher % fat in the arm (p = 0.015), leg (p = 0.034) and as total (p = 0.017), lower haemoglobin (p = 0.002) and creatinine levels (p = 0.008), and higher CD4 cell counts (p = 0.009) (Mann–Whitney test). BMI, body mass index; DEXA, Dual-energy X-ray absorptiometry; HOMA, Homeostasis Model Assessment; VACS, Veterans Ageing Cohort Study (DOCX 94 kb)

Values of cholesterol at baseline (BL) and week-12 (W12) in patients untreated with statins. Values are expressed as median (Q1-Q3). W12 values were compared to BL values by the Wilcoxon matched-pairs signed rank test. (DOCX 46 kb)

Performance of participants during the 12Â weeks of training in the walk group divided by gender. Values as expressed as median (Q1-Q3). a. The median HRmean value was calculated for each participant through all his/her walking sessions, to derive the median HRmean value of all participants. HR, heart rate. (DOCX 38 kb)

Additional body composition and laboratory values at baseline (BL) and week-12 (W12). Values are expressed as median (Q1-Q3). W12 values were compared to BL values by the Wilcoxon matched-pairs signed rank test. BMI, body mass index; DEXA, Dual-energy X-ray absorptiometry; HOMA, Homeostasis Model Assessment; VACS, Veterans Ageing Cohort Study (DOCX 100 kb)

Introduction: The RESPIRA study (EU Funded-Italia-Malta) was performed in the district of Gela (Southern Sicily) and Malta in 2012/3 in 10-15 years old schoolchildren (n=2,047) by means of standard respiratory questionnaires (ISAAC),and clinical exam of case and control subjects. Aims and Objectives: To measure total serum IgE levels in cases (n=127, males=45) and controls (n=142, Males=64), specific IgEs in cases (n=90, Males=53), and controls (n=82, Males=38). Materials and Methods: Cases reported wheeze and/or use of asthma medications in the last 12 months, while controls answered negatively to both and rhinitis. Results: Total Serum IgE (n=269) was raised (>144 IU/l) in 70/127 cases vs 25/142 controls (p=.0001). In males 30/76 of cases vs 11/64 controls (p=.005), while in females cases 15/51 vs 14/78, (p=.14). Positive specific IgE (>0.34 U/ml) was noted for at least one allergen in 54/90 cases vs 30/82 controls (p=.002). House dust mite (HDM) was positive in 27/90 cases vs 12/82 controls (p=.0001), Parietaria positive 12/90 cases vs 2/82 controls (p=.011), cat 22/90 vs 13/82 controls (p=.19), while in male subgroup 14/53 of cases were positive vs 4/38 controls (p=.06). Dog 9/90 cases vs 8/82 controls (NS). Olive positive in 13/90 cases vs 8/82 controls (NS). Alternaria positive in 6/90 cases vs 2/82 controls (NS). Goldenrod Solidago Virgaurea positive in 2/90 cases vs 1/82 controls, and Cladosporium 0/90 cases, 1/82 controls. Conclusion: Positive Total serum IgE and specific IgE to HDM and Parietaria in all patients and cat in males only were higher in cases than in controls., peer-reviewed

A biorefinery should ideally integrate biomass conversion processes to produce a range of fuels, power, materials, and chemicals from biomass. The term biorefinery is derived both from the raw material feedstock which is renewable biomass and also from the bioconversion processes often applied in the treatment and processing of the raw materials. Renewable sources are the basis of the alternative energy, fuels and compound obtained in a biorefinery-based way to meet the energy demand in a world where petrol fuels are becoming scarce and more expensive. The current biorefinery schemes are mainly based on hexose sugars (first generation) or lignocellulosic-based material (second generation) as feedstocks and depend on microbial cell factories for obtaining the products of interest. In most cases, the substrate utilization is not optimized jet: at the end of the process in addition to the main product, by-products such as huge quantity of lignin and some C5 sugars (deriving from incomplete utilization of lignocellulose) but also proteins (mainly the exhausted biomass at the end of the production) are still available. Focusing the attention on proteins, they are nowadays mainly absorbed as animal feed, but the quantities produced exceed the demand. Considering the implication related to GMO utilization and the increasing number of biorefineries, it can be stated that proteins could represent an interesting Abstract - 2 - “no-cost” starting material to produce more biofuel, bioproducts or biopower in a biorefinery-based way. This study has the aim to present some examples of production by using the protein side-stream of a biorefinery. In particular, we report the possible destiny of two aminoacids released from said proteins: glycine, to obtain butanol and isobutanol, and glutamate, to produce succinic acid. Butanol represents the most feasible alternative to gasoline, but, differently from ethanol, microbial fermentations are not enough developed to sustain the market demand. The natural producers, different species of the Gram positive group of Clostridia, present many and different limitations which are challenging the scientific communities in finding alternative hosts. Among the different alternative hosts, yeasts can present attractive features, being the most relevant the existence of industrial plants based on Saccharomyces cerevisiae fermentation for the production of ethanol. Up to now, in S. cerevisiae the production of butanol has been obtained by expressing heterologous genes from Clostridia. The best reported butanol titer is only 2.5 mg/L, probably mainly depending on problems related to the overexpression of heterologous prokaryotic enzymes. On the contrary, the isobutanol production is obtained at high level (0.63 g/L) in S. cerevisiae by using and optimizing an endogenous valine degradation pathway. We present an alternative and novel way to produce butanol and isobutanol using glycine as substrate, through a new identified and Abstract - 3 - biochemically characterized pathway. Starting from 15 g/L of glycine we have obtained 92 mg/L of butanol and 58 mg/L of isobutanol. Considering that the pathway has been just discovered and no optimization has been designed or applied, the obtained results has to be considered extremely positive. Since one of the most common stresses that microorganisms have to face during productions is the toxicity of the final product, we investigated also the possibility to use the glycine as protective agent for the cells. This idea originated from different studies that explain how the use of aminoacids, among which glycine, can help the cells to better respond to different stresses, such as high ethanol concentration and hydrogen peroxide. As never reported in literature, we have notably found that butanol causes a sort of oxidative stress, peroxidising the membrane lipids. Moreover, we have demonstrated that glycine can help the cells to better tolerate the presence of hydrogen peroxide, acetic acid, ethanol and butanol. The succinic acid occurs naturally in humans, animals, plants, and microorganisms as TCA cycle intermediate. Its use as a precursor to produce many important commodity chemicals used to make a wide assortment of products such as solvents, fiber and polymer production make it one of the most important relevant bulk chemicals. The microbial production of succinic acid via reductive TCA cycle using engineered Saccharomyces cerevisiae is nowadays on the market commercialised by Reverdia (Cassano Spinola, Italy). Abstract - 4 - Here we investigated the possibility to exploiting the glutamate degradation pathway through the γ-aminobutyric acid (GABA) intermediate as an additional way of obtaining succinate when a source of glutamate is feeded to the cells. Since this pathway, also called GAD/GABA shunt, is transcriptionally controlled, we transcriptionally up-regulated it through the constitutive overexpression of the involved genes, GAD1, UGA1 and UGA2. Even if the pathway was clearly overexpressed, as revealed by transcriptional analysis, the obtained amount of succinic acid did not reflect this modification. Further investigating the possible reason of that, we have demonstrated that, despite the higher transcription levels, the corresponding enzymatic activities of the pathway did not increased. We can argue that posttranslational regulations occur in the pathway, what at the moment impaired the initial aim of our work. However, the data add new and relevant information for the comprehension of the role and the regulation of this pathway in S. cerevisiae. It cannot be excluded that the production of succinic acid in yeasts might be in a future optimized by exploiting the GAD/GABA pathway, once it will be fully characterized. We have finally investigated the possibility to produce succinic acid in a strain of S. cerevisiae deleted in the gene encoding the enzyme glutamate synthase, GLT1. During this study we have tested different growing conditions, since the target gene of deletion is located in a critical node of the nitrogen sensing and utilization, which is related to cellular plasticity and adaptability in perturbed environment. We could demonstrate that Abstract - 5 - when cells are growing in low salts concentration medium, the deleted strain compared to wild type strain is able to accumulate more reduced products, such as succinic acid, malate/fumarate, glycerol and ethanol, indicating a sort of necessity to reduce the excess of NAD(P)H generated by the GLT1 deletion. Regarding this preliminary study, deeper investigations have to be performed to find a fine tuned way to intentionally redirect the metabolic flux and obtain the product of interest, which in turn could be succinic acid, but also malic or fumaric acid, or again ethanol, being this last the product that has the responsibility to prove that bio-based second generation processes can effectively and positively have an impact on our future society.

Cigarette smoke is the main source of indoor chemical and toxic elements. Cadmium (Cd), Thallium (Tl), Lead (Pb) and Antimony (Sb) are important contributors to smoke-related health risks. Data on the association between Rare Earth Elements (REE) Cerium (Ce) and Lanthanum (La) and domestic smoking are scanty. To evaluate the relationship between cigarette smoke, indoor levels of PM2.5 and heavy metals, 73 children were investigated by parental questionnaire and skin prick tests. The houses of residence of 41 "cases" and 32 "controls" (children with and without respiratory symptoms, respectively) were evaluated by 48-hours PM2.5 indoor/outdoor monitoring. PM2.5 mass concentration was determined by gravimetry; the extracted and mineralized fractions of elements (As, Cd, Ce, La, Mn, Pb, Sb, Sr, Tl) were evaluated by ICP-MS. PM2.5 and Ce, La, Cd, and Tl indoor concentrations were higher in smoker dwellings. When corrected for confounding factors, PM2.5, Ce, La, Cd, and Tl were associated with more likely presence of respiratory symptoms in adolescents. We found that: i) indoor smoking is associated with increased levels of PM2.5, Ce, La, Cd, and Tl and ii) the latter with increased presence of respiratory symptoms in children

Supplemental Digital Content is available in the text, Background: Cerebrospinal fluid (CSF) viral escape is a concern in ritonavir-boosted protease inhibitors monotherapy. The aim was to assess HIV-RNA, biomarkers of immune activation and neurodegeneration, and atazanavir concentrations in CSF of patients on successful long-term atazanavir/ritonavir (ATV/r) monotherapy. Methods: This is a substudy of the multicentric, randomized, open-label, noninferiority trial monotherapy once a day with atazanavir/ritonavir (NCT01511809), comparing the ongoing ATV/r along with 2 nucleoside retrotranscriptase inhibitors (NRTIs) regimen to a simplified ATV/r monotherapy. Patients with plasma HIV-RNA < 50 copies/mL after at least 96 study weeks were eligible. We assessed HIV-RNA, soluble (s)CD14, sCD163, CCL2, CXCL10, interleukin-6, and YKL40 by enzyme-linked immunosorbent assay; neopterin, tryptophan, kynurenine, and neurofilament by immunoassays; and ATV concentrations by liquid chromatography–mass spectrometry in paired plasma and CSF samples. Variables were compared with Wilcoxon rank-sum or Fisher exact test, as appropriate. Results: HIV-RNA was detected in the CSF of 1/11 patients on ATV/r monotherapy (114 copies/mL), without neurological symptoms, who was successfully reintensified with his previous 2NRTIs, and in none of the 12 patients on ATV/r + 2NRTIs. CSF biomarkers and ATV concentrations did not differ between the 2 arms. Conclusions: CSF escape was uncommon in patients on long-term ATV/r monotherapy and was controlled with reintensification.