Individual particle SEM-EDS analysis of atmospheric aerosols in rural, urban, and industrial sites of Central Italy.

PM10 samples were collected simultaneously at three representative areas (urban, industrial, and rural areas). Their morphology and elemental composition were determined by scanning electron microscopy coupled with energy-dispersive analysis (SEM-EDS). Twenty-four chemical parameters (C, O, Na, Mg, Al, Si, P, Cd, Cl, K, Ca, S, Sn, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, W, and Pb) were determined and three morphological parameters (area, roundness, and fractal dimension) were measured by Image Pro Analyzer 6.3. The particles were classified into ten groups based on morphology and elemental composition: Ca-rich and metal particles, soot aggregates, cenosphere, alumosilicates, sea salt, calcium sulfate, spherical particles of iron, biological carbonaceous particles, and various. Particles of natural origin were predominantly found in the coarse size fraction and particles of anthropogenic origin in the fine size fraction. The greatest contribution to particulate matter belonged to aluminum-silicates and calcium-rich particles. The cenosphere were recognized only in the coastal urban site, while all the other particles were present in each site. The coastal industrial site was characterized by the prevalence of alumosilicates and Ca-rich particles, due to construction activity in this site during the sampling period (movement of vehicles, transport of terrigenous materials, and use of construction products). The coastal urban site was characterized by a higher amount of soot and by the presence of cenosphere, due to the presence of vehicular traffic.

Source apportionment of PM(2.5) in the harbour-industrial area of Brindisi (Italy): identification and estimation of the contribution of in-port ship emissions.

Harbours are important for economic and social development of coastal areas but they also represent an anthropogenic source of emissions often located near urban centres and industrial areas. This increases the difficulties in distinguishing the harbour contribution with respect to other sources. The aim of this work is the characterisation of main sources of PM2.5 acting on the Brindisi harbour-industrial area, trying to pinpoint the contribution of in-port ship emissions to primary and secondary PM2.5. Brindisi is an important port-city of the Adriatic Sea considered a hot-spot for anthropogenic environmental pressures at National level. Measurements were performed collecting PM2.5 samples and characterising the concentrations of 23 chemical species (water soluble organic and inorganic carbon; major ions: SO4(2-), NO3(-), NH4(+), Cl(-), C2O4(2-), Na(+), K(+), Mg(2+), Ca(2+); and elements: Ni, Cu, V, Mn, As, Pb, Cr, Sb, Fe, Al, Zn, and Ti). These species represent, on average, 51.4% of PM2.5 and were used for source apportionment via PMF. The contributions of eight sources were estimated: crustal (16.4±0.9% of PM2.5), aged marine (2.6±0.5%), crustal carbonates (7.7±0.3%), ammonium sulphate (27.3±0.8%), biomass burning-fires (11.7±0.7%), traffic (16.4±1.7 %), industrial (0.4±0.3%) and a mixed source oil combustion-industrial including ship emissions in harbour (15.3±1.3%). The PMF did not separate the in-port ship emission contribution from industrial releases. The correlation of estimated contribution with meteorology showed directionality with an increase of oil combustion and sulphate contribution in the harbour direction with respect to the direction of the urban area and an increase of the V/Ni ratio. This allowed for the use of V as marker of primary ship contribution to PM2.5 (2.8%+/-1.1%). The secondary contribution of oil combustion to non-sea-salt-sulphate, nssSO4(2-), was estimated to be 1.3 µg/m(3) (about 40% of total nssSO4(2-) or 11% of PM2.5).

Source apportionment of size-segregated atmospheric particles based on the major water-soluble components in Lecce (Italy).

Atmospheric aerosols have potential effects on human health, on the radiation balance, on climate, and on visibility. The understanding of these effects requires detailed knowledge of aerosol composition and size distributions and of how the different sources contribute to particles of different sizes. In this work, aerosol samples were collected using a 10-stage Micro-Orifice Uniform Deposit Impactor (MOUDI). Measurements were taken between February and October 2011 in an urban background site near Lecce (Apulia region, southeast of Italy). Samples were analysed to evaluate the concentrations of water-soluble ions (SO4(2-), NO3(-), NH4(+), Cl(-), Na(+), K(+), Mg(2+) and Ca(2+)) and of water-soluble organic and inorganic carbon. The aerosols were characterised by two modes, an accumulation mode having a mass median diameter (MMD) of 0.35 ± 0.02 µm, representing 51 ± 4% of the aerosols and a coarse mode (MMD=4.5 ± 0.4 µm), representing 49 ± 4% of the aerosols. The data were used to estimate the losses in the impactor by comparison with a low-volume sampler. The average loss in the MOUDI-collected aerosol was 19 ± 2%, and the largest loss was observed for NO3(-) (35 ± 10%). Significant losses were observed for Ca(2+) (16 ± 5%), SO4(2-) (19 ± 5%) and K(+) (10 ± 4%), whereas the losses for Na(+) and Mg(2+) were negligible. Size-segregated source apportionment was performed using Positive Matrix Factorization (PMF), which was applied separately to the coarse (size interval 1-18 µm) and accumulation (size interval 0.056-1 µm) modes. The PMF model was able to reasonably reconstruct the concentration in each size-range. The uncertainties in the source apportionment due to impactor losses were evaluated. In the accumulation mode, it was not possible to distinguish the traffic contribution from other combustion sources. In the coarse mode, it was not possible to efficiently separate nitrate from the contribution of crustal/resuspension origin.

Metal and non-metal micro-pollutants in cultivated soils of Lecce and Brindisi districts (Apulia, southern Italy).

To estimate the presence of metal and non-metal micro-pollutants in the soils of the Lecce and Brindisi districts, an analytical study has been carried out on samples of surface soils taken from agricultural areas. The research has concerned the determination of the following micro-pollutants: Cu, Ni, Cr, Zn, Pb, Be, Cd, As, Hg, Sb e Tl. Statistical techniques, such as Principal Component Analysis and Clustering Analysis, have been utilised to examine the correlations among the different parameters and to define contamination areas. The results show that the amount of micro-pollutants in the superficial stratum of the examined soils is in the range permitted by the regulations in force, with the exception of arsenic and thallium. Arsenic concentrations are near to the maximum admissible value, while thallium concentrations are in 56% of the samples higher than the admissible value both in the soil and underground. The most significant parameter from a toxicological point of view is thallium, which has its maximum concentration in the soils located near the industrial area of Brindisi.

Sea water contamination in underground waters of salento (Southern Italy).

In the present work, a study of a physico-chemical characterisation of underground waters, utilised for agriculture and human use in the Lecce district (Southern Italy) has been reported. The aim of the work has been to define the quality of underground waters in the different areas and to value salt contamination due to seawater intrusion. Statistical techniques, such as Principal Component Analysis (PCA) and Cluster Analysis (CA), have been utilised to examine the correlations among the different parameters and to define contamination areas. The results have shown a high salt contamination in artesian wells of the Ionian Sea coast.

Specific combinations of zein genes and genetic backgrounds influence the transcription of the heavy-chain zein genes in maize opaque-2 endosperms.

The transcript levels of heavy-chain zein genes (zH1 and zH2) and the occurrence of the zH polypeptides in different opaque-2 (o2) lines were investigated by RNA-blot analyses and by sodium dodecylsulfate-polyacrylamide gel electrophoresis or two-dimensional gel electrophoresis protein fractionations. Four mutant alleles o2R, o2T, o2It, and o2-676 introgressed into different genetic backgrounds (GBs) were considered. The mono-dimensional gel electrophoresis zein pattern can be either conserved or different among the various GBs carrying the same o2 allele. Likewise, in the identical GB carrying different o2 alleles, the zein pattern can be either conserved or differentially affected by the different mutant allele. Zein protein analysis of reciprocal crosses between lines with different o2 alleles or the same o2 showed in some case a more than additive zH pattern in respect to the o2 parent lines. Electrophoretic mobility shift assay approaches, with O2-binding oligonucleotide and endosperm extracts from the above o2 lines, failed to reveal o2-specific retarded band in any of the o2 extracts. The results suggest that the promoter of some zH1 and zH2 contains motif(s) that can respond to factors other than O2.

The activity of the maize Opaque2 transcriptional activator is regulated diurnally.

The maize (Zea mays L.) Opaque2 (O2) protein is an endosperm-specific transcriptional activator whose DNA-binding activity is regulated diurnally by a phosphorylation/dephosphorylation mechanism. We show that the O2 transcript undergoes pronounced oscillations during the day-night cycle. The highest level of the O2 message is present at midday and the lowest level at midnight. The level of O2 transcript follows a diurnal rhythm that appears controlled by the circadian clock. Two different endosperm-expressed DNA-binding proteins, PBF (prolamin box-binding factor) and OHP1 (O2-heterodimerizing protein 1), were also analyzed. While the PBF message levels oscillate diurnally, the steady-state levels of OHP1 transcript were constant through the day and night. We present data showing that the seed is not directly involved in the perception of the light signal, but presumably responds to diurnal fluxes of nutrients into the endosperm. Moreover, we show that the O2 protein is not involved in the regulation of its own transcript levels. These data indicate that O2 activity is down-regulated at night by both a reduction in O2 transcript and by hyperphosphorylation of residual O2 protein, and suggest that regulatory gene activity during endosperm development may be acutely sensitive to a diurnal signal(s) emanating from the plant and passing into the developing seeds.

Phosphorylation of Opaque2 changes diurnally and impacts its DNA binding activity.

In the maize endosperm, the Opaque2 (O2) basic leucine zipper transcriptional activator regulates the expression of a subset of the zein seed storage protein gene family. Immunodetection of wild-type or mutant O2 polypeptides fractionated by SDS-PAGE resolved a closely spaced doublet migrating in the 68- to 72-kD range, whereas by using isoelectric focusing, seven to nine isoforms were detected for each allele. Phosphatase treatment simplified the protein patterns to a single band corresponding to the nonphosphorylated component. In vivo and in vitro labeling confirmed that O2 can be phosphorylated. In protein gel blots probed with DNA, only the nonphosphorylated and hypophosphorylated O2 polypeptides were able to bind an oligonucleotide containing the O2 binding sequence. Upon in situ dephosphorylation of the focused isoforms by phosphatase treatment of the isoelectric focusing filter, the hyperphosphorylated forms acquired DNA binding activity. The ratio among the various isoforms remained constant throughout the developmental stages of endosperm growth but changed from daytime to nighttime, with a significant increase of the hyperphosphorylated forms during the night period. These results indicate that O2 exists in vivo as a pool of differently phosphorylated polypeptides and demonstrate that O2 DNA binding activity is modulated by a phosphorylation/dephosphorylation mechanism that appears to be influenced by environmental conditions.

[The role of Ca(2+)-ATPase and oxygen radical in reperfusion injury of rat liver].

The effects of ischemia and reperfusion with and without oxygen radical scavengers and xanthine oxidase inhibitors on Ca(2+)-ATPase activity were examined in the rat liver of 5 min ischemia followed by 5 and 10 min reperfusion. Ischemia was produced by the ligation of right hepatic artery and right portal vein. Superoxide dismutase, catalase and allopurinol were administered by subcutaneous injection of 60,000U/kg, 90,000U/kg and 200mg/kg, respectively before ligation. Reaction products of Ca(2+)-ATPase were morphometrically analyzed by RUZEX IIIU. Histochemically, Ca(2+)-ATPase activities were demonstrated on plasma membrane of liver cells, bile canaliculi and Kupffer cells involving mitochondria in liver cells of control rats. Ca(2+)-ATPase activities were depressed in the central lobes of liver after 5 min ischemia followed by 5 and 10min reperfusion. However, the activities of Ca(2+)-ATPase were not depressed by addition of oxygen radical scavengers and xanthine oxidase inhibitor before ischemia. These results suggest that oxygen free radicals may influence Ca(2+)-ATPase activity and contribute to liver cell damage due to ischemia-reperfusion.

Endocardial mapping of ventricular tachycardia in the intact human heart. II. Evidence for multiuse reentry in a functional sheet of surviving myocardium.

OBJECTIVE: The purpose of this study was to obtain improved detection and characterization of reentrant circuits in the infarcted human ventricle. BACKGROUND: The return path of reentrant ventricular arrhythmias usually is not manifested in clinical mapping studies but is thought to be formed by isolated bundles of surviving myocytes whose presence is difficult to detect by standard recording techniques. METHODS: We obtained simultaneous unipolar and high gain bipolar recordings using a left ventricular endocardial balloon array in 10 patients with chronic ischemic heart disease undergoing intraoperative mapping of ventricular tachycardia. RESULTS: Three patients demonstrated seven separate ventricular tachycardias that utilized a return tract that was manifested on up to 20% of all left ventricular electrode sites. The recordings suggested an extensive sheet of surviving myocardial fibers with multiple entry and exit points allowing for different reentrant paths at different times all in the same heart. In one patient, five different ventricular tachycardias could be induced, four of which utilized such a sheet. Two of these tachycardias had the same exit point (site of origin) but two different entry points with a long and short return path resulting in long and short tachycardia cycle lengths. The same sheet sustained another tachycardia with one entry and two exit points resulting in two separate "sites of origin" on the endocardium. Such sheets also were seen to insert into the left bundle system. In one patient portions of the sheet could be detected epicardially. CONCLUSION: The existence of such a structure of surviving myocardium with functional pleomorphism may account for unexplained changes in tachycardia cycle length, epicardial entrainment and spontaneous morphologic changes during ventricular tachycardia.

Ventricular tachycardia after surgical repair of tetralogy of Fallot: results of intraoperative mapping studies.

OBJECTIVES: Four patients with previous repair of tetralogy of Fallot and ventricular tachycardia underwent map-guided surgery to ablate the arrhythmias. BACKGROUND: Although patients with repaired tetralogy of Fallot are at increased risk of sudden death due to ventricular tachycardia, little is known of the origin and mechanism of this arrhythmia. METHODS: A customized right ventricular balloon with 112 electrodes was used to record endocardial activation and, where possible, simultaneous epicardial recordings were obtained with a sock electrode array. Three patients had an aneurysm of the right ventricular outflow tract and one had a septal aneurysm. All had moderate to severe pulmonary valve insufficiency. Preoperative electrophysiologic study demonstrated inducible rapid (cycle length 180 to 300 ms) hemodynamically unstable monoform ventricular tachycardias. RESULTS: Intraoperatively, five different tachycardias (two in one patient) were induced and mapped. The sites of earliest activation were located in the subendocardium of the right ventricular outflow tract in all, but they varied widely among the septum, free wall and parietal band and could not be identified by visible scar. All were due to a macroreentrant circuit initiated by a critical delay in activation beyond a functional arc of block. Two patients treated by cryoablation while the heart was beating and perfused at normal temperature had inducible ventricular tachycardia postoperatively. In the two subsequent patients, the application of cryoablation under anoxic cardiac arrest resulted in noninducibility of arrhythmia. CONCLUSIONS: Ventricular tachycardia in tetralogy of Fallot in these four patients was caused by macroreentry in the right ventricular outflow tract. Surgical success depends on detailed mapping and cryoablation under anoxic cardiac arrest. In patients at risk of sudden death, map-directed surgery may offer distinct advantages over either implantable devices or drug therapy.

A carrot cell variant temperature sensitive for somatic embryogenesis reveals a defect in the glycosylation of extracellular proteins.

The temperature-sensitive carrot cell variant ts11c, arrested in somatic embryogenesis after the globular stage, was characterized. The sensitivity to a shift from 24 degrees C (permissive temperature) to 32 degrees C (non-permissive temperature) is greatest at the globular stage of embryogenesis, while cells proliferating in unorganized fashion and plantlets are not affected. Embryogenesis in ts11c is also arrested at the permissive temperature by replacement of conditioned culture medium with fresh medium. The timing of sensitivity of ts11c to medium replacement coincides with the sensitivity to temperature shift. Both sensitivities are recessive in somatic hybrids between ts11c and wild-type cells. Extracellular glycoproteins synthesized by ts11c at the non-permissive temperature contain much less fucose than those synthesized by the wild type. The glycoproteins synthesized by the variant under non-permissive conditions do not accumulate at the periphery of the embryo, as their wild-type counterparts do, but instead show a diffuse distribution throughout the embryo. The defect in ts11c can be fully complemented by the addition of extracellular wild-type proteins. A revertant of ts11c was isolated that simultaneously reacquired temperature insensitivity and normal glycosylation ability. Collectively, these observations indicate that ts11c is not able to perform proper glycosylation at the non-permissive temperature and suggest that the activity of certain extracellular proteins, essential for the transition of globular to heart stage somatic embryos, depends on the correct modification of their oligosaccharide side-chains.

Synthesis of Lectin-Like Protein in Developing Cotyledons of Normal and Phytohemagglutinin-Deficient Phaseolus vulgaris.

The genome of the common bean Phaseolus vulgaris contains a small gene family that encodes lectin and lectin-like proteins (phytohemagglutinin, arcelin, and others). One of these phytohemagglutinin-like genes was cloned by L. M. Hoffman et al. ([1982] Nucleic Acids Res 10: 7819-7828), but its product in bean cells has never been identified. We identified the product of this gene, referred to as lectin-like protein (LLP), as an abundant polypeptide synthesized on the endoplasmic reticulum (ER) of developing bean cotyledons. The gene product was first identified in extracts of Xenopus oocytes injected with either cotyledonary bean RNA or LLP-mRNA obtained by hybrid-selection with an LLP cDNA clone. A tryptic map of this protein was identical with a tryptic map of a polypeptide with the same SDS-PAGE mobility detectable in the ER of bean cotyledons pulse-labeled with either [(3)H]glucosamine or [(3)H]amino acids, both in a normal and in a phytohemagglutinin-deficient cultivar (cultivars Greensleeves and Pinto UI 111). Greensleeves LLP has M(r) 40,000 and most probably has four asparagine-linked glycans. Pinto UI 111 LLP has M(r) 38,500. Unlike phytohemagglutinin which is a tetramer, LLP appears to be a monomer by gel filtration analysis. Incorporation of [(3)H]amino acids indicates that synthesis of LLP accounts for about 3% of the proteins synthesized on the ER, a level similar to that of phytohemagglutinin.

A nuclear mutant of Saccharomyces cerevisiae deficient in mitochondrial DNA replication and polymerase activity.

We have isolated a thermosensitive mutant which is transformed into a population of cells devoid of mitochondrial DNA (rho 0 cells) at 35 degrees C and is deficient in mitochondrial (mt) DNA polymerase activity. A single recessive nuclear mutation (mip1) is responsible for rho 0 phenotype and mtDNA polymerase deficiency in vitro. At 25 degrees C (or 30 degrees C) a dominant suppressor mutation (SUP) masks the deficiency in vivo. The meiotic segregants (mip1 sup) which do not harbor the suppressor have a rho 0 phenotype both at 25 and 35 degrees C. They have no mtDNA polymerase activity, in contrast with MIP rho 0 mutants of mitochondrial inheritance which do exhibit mtDNA polymerase activity. In the thermosensitive mutant (mip1 SUP), the replication of mtDNA observed in vivo at 30 degrees C is completely abolished at 35 degrees C. In the meiotic segregants (mip1 sup), no mtDNA replication takes place at 30 and 35 degrees C. The synthesis of nuclear DNA is not affected. DNA polymerases may have replicative and/or repair activity. There is no evidence that mip mutants are deficient in mtDNA repair. In contrast the MIP gene product is strictly required for the replication of mtDNA and for the expression of the mtDNA polymerase activity. This enzyme might be the replicase of mtDNA.

ALG/alg: a single gene controlling the utilization of lactate in the presence of antimycin in the yeast Saccharomyces cerevisiae.

A strain dependent growth on lactate in the presence of antimycin A (AA) has been observed - the strain D261 can grow on lactate and AA, whereas in the strain K8/6C antimycin A prevents the utilization of lactate and the induction of LDH.Genetic analysis demonstrates that growth on lactate in the presence of AA segregates from D261 as a single nuclear factor which we indicate by ALG1 and alg1 in its dominant and recessive states. alg1 complements the gene(s) which give(s) rise to the same phenotype in K8/6C.The analysis of the regulation by lactate of LDH in the absence and presence of AA and in rho (-) cells shows that growth on lactate and antimycin A is not corretated with the induction by lactate of LDH.

Mitochondrial NAD, L-lactate dehydrogenase and NAD, D-lactate dehydrogenase in the yeast Saccharomyces cerevisiae.

Mitochondrial NAD-linked L- and D-lactate dehydrogenase activities have been found in the yeast Saccharomyces cerevisiae grown at high (3%) but were absent at low (0.6%) glucose concentrations. The inhibition of mitochondrial protein synthesis by chloramphenicol and of primary respiration by antimycin A determines the appearance of the two activities even at low (0.6%) glucose concentration. Two respiratory deficient strains belonging respectively to the mit- class (which maintains mitochondrial protein synthesis) and to the rho- class (which loses mitochondrial protein synthesis) display the activities even at low (0.6%) glucose concentration. L- and D-lactate have been detected in the growth medium when the cultures had been undertaken at high glucose concentrations, but were absent at low glucose concentrations.