A combined chemical/size fractionation approach to study winter/summer variations, ageing and source strength of atmospheric particles.

We studied the size distribution of ions (Cl-, NO3-, SO4=, Na+, NH4+, K+, Mg++, Ca++) and elements (As, Ba, Cd, Co, Cs, Cu, Fe, Li, Mn, Ni, Pb, Rb, Sb, Se, Sn, Sr, Ti, Tl, V, Zn) during the winter and summer seasons of seven consecutive years (2008-2014) in an area of the Po Valley (Northern Italy) characterised by industrial, agricultural and urban settings. The study included the collection and analysis of 41 series of size-segregated samples (MOUDI sampler, 10 stages, cut sizes from 0.18 to 18 µm). Ions were analysed by ion chromatography; elemental analysis was carried out by ICP-MS, by applying a chemical fractionation method able to increase the selectivity of PM source tracers. Our results indicate that important winter/summer variations occurred in both the concentration and size distribution of most PM components. These variations were explained in terms of variations in the strength of the prevailing sources of each component. The contribution of biomass burning for domestic heating was highlighted by the well-known tracer K+ but also by the soluble fraction of Rb, Cs and Li. Biomass burning contribution to atmospheric PM was mostly contained in the fine fraction, with a broad size-distribution from 0.18 to 1.8 µm. This source also appreciably increased the concentration of other elements in fine PM (As, Cd, Co, Mn, Pb, Sb, Sn). A few PM components (tracers of sea-spray, brake lining and some industries) did not show marked seasonal variations in concentration and size distribution. However, during winter, for brake lining and industry tracers we observed an upward shift in the dimension of fine particles and a downward shift in the dimension of coarse particles, due to the ageing of the air masses.

Particulate matter concentration and chemical composition in the metro system of Rome, Italy.

Air quality at the main station of the metro system of Rome (Termini hub) has been characterized by the point of view of particulate matter (PM) concentration and chemical composition. Indoor air in different environments (underground train platform and shopping center, metro carriages with and without air conditioning system) has been studied and compared with outdoor air at a nearby urban site. Air quality at the railway station, located outdoor at surface level, has been also considered for comparison. PM chemical characterization included ions, elemental carbon, organic carbon, macro-elements, and the bio-accessible and residual fractions of micro- and trace elements. Train platform and carriages without air conditioning resulted to be the most polluted environments, with indoor/outdoor ratio up to two orders of magnitude for many components. PM mass concentration was determined on filter membranes by the gravimetric procedure as well as from the optical particle counter (OPC) number concentration measurements. The OPC results, taken with the original calibration factor, were below 40 % of the value obtained by the gravimetric measurements. Only a chemical and morphological characterization of the collected dust could lead to a reconciliation of the results yielded by the two methods. Macro-components were used to estimate the strength of the main macro-sources. The most significant contribution is confirmed to derive from wheels, rails, and brakes abrasion; from soil re-suspension (over 50 % at the subway platform); and from organics (about 25 %). The increase in the concentration of elements was mostly due to the residual fraction, but also the bio-accessible fraction showed a remarkable enrichment, particularly in the case of Ba, Zn, Cd, and Ni.

Recognition of B and T cell epitopes by cattle immunized with a synthetic peptide containing the major immunogenic site of VP1 FMDV 01 Campos.

The precise location of B and T cell epitopes have been established in a peptide containing the major immunogenic site (residues 135-160) of FMDV strain 01 Campos (01C) VP1. The peptide (p135-160), administered free or conjugated to bovine serum albumin, induced complete protection in guinea pigs and a strong neutralizing antibody (NAb) response in cattle. Using a set of partially overlapping peptides it was shown that although several B cell epitopes were distributed along the p135-160, the residues responsible for the induction of NAb were restricted to the amino acids 135-144. In addition, bovines immunized with the p135-160 showed a strong proliferative response to this peptide but a very poor response against purified virus. Two T-epitopes were located in the 135-160 sequence: one on the region 135-144 and the other in the region 150-160. The immunodominance of these two T cell epitopes was confirmed in cattle immunized with inactivated virus vaccines.

[Comparison of various methods for the measurement of the humoral immune response in mice vaccinated with aphthous fever virus]. / Comparación de distintos métodos para la medición de la respuesta inmune humoral de ratones vacunados con el virus de la fiebre aftosa.

Kinetics of the humoral immune primary response and seven-day secondary response of adult CF1 mice to FMDV O1 Campos adjuvanted in aluminium hydroxide-saponin (AHS) or in oil emulsion (OE) were evaluated by means of ELISA and passive hemagglutination (PH). Analysis of the response to AHS vaccine showed that ELISA measured maximal titres of primary response at 23 days post-vaccination (dpv), and at day 17 of secondary response, while PH detected maximal titres for primary as well as secondary response around day 60 pv. Mice immunized with OE vaccine studied by ELISA presented a plateau of primary response around 40 dpv, while secondary response was maximal around 80 dpv. The same sera tested by PH showed the highest titres for primary response on day 50 pv and secondary response was maximal on day 40 pv. A criterion for the evaluation of vaccination efficiency in the murine model is proposed based on the methods employed in the determination of antibody level.

Comparación de distintos métodos para la medición de la respuesta inmune humoral de ratones vacunados con el virus de la fiebre aftosa / [Comparison of various methods for the measurement of the humoral immune response in mice vaccinated with aphthous fever virus]

Kinetics of the humoral immune primary response and seven-day secondary response of adult CF1 mice to FMDV O1 Campos adjuvanted in aluminium hydroxide-saponin (AHS) or in oil emulsion (OE) were evaluated by means of ELISA and passive hemagglutination (PH). Analysis of the response to AHS vaccine showed that ELISA measured maximal titres of primary response at 23 days post-vaccination (dpv), and at day 17 of secondary response, while PH detected maximal titres for primary as well as secondary response around day 60 pv. Mice immunized with OE vaccine studied by ELISA presented a plateau of primary response around 40 dpv, while secondary response was maximal around 80 dpv. The same sera tested by PH showed the highest titres for primary response on day 50 pv and secondary response was maximal on day 40 pv. A criterion for the evaluation of vaccination efficiency in the murine model is proposed based on the methods employed in the determination of antibody level.

Comparación de distintos métodos para la medición de la respuesta inmune humoral de ratones vacunados con el virus de la fiebre aftosa / [Comparison of various methods for the measurement of the humoral immune response in mice vaccinated with aphthous fever virus]

Kinetics of the humoral immune primary response and seven-day secondary response of adult CF1 mice to FMDV O1 Campos adjuvanted in aluminium hydroxide-saponin (AHS) or in oil emulsion (OE) were evaluated by means of ELISA and passive hemagglutination (PH). Analysis of the response to AHS vaccine showed that ELISA measured maximal titres of primary response at 23 days post-vaccination (dpv), and at day 17 of secondary response, while PH detected maximal titres for primary as well as secondary response around day 60 pv. Mice immunized with OE vaccine studied by ELISA presented a plateau of primary response around 40 dpv, while secondary response was maximal around 80 dpv. The same sera tested by PH showed the highest titres for primary response on day 50 pv and secondary response was maximal on day 40 pv. A criterion for the evaluation of vaccination efficiency in the murine model is proposed based on the methods employed in the determination of antibody level.

Comparación de distintos métodos para la medición de la respuesta inmune humoral de ratones vacunados con el virus de la fiebre aftosa. / [Comparison of various methods for the measurement of the humoral immune response in mice vaccinated with aphthous fever virus]

Kinetics of the humoral immune primary response and seven-day secondary response of adult CF1 mice to FMDV O1 Campos adjuvanted in aluminium hydroxide-saponin (AHS) or in oil emulsion (OE) were evaluated by means of ELISA and passive hemagglutination (PH). Analysis of the response to AHS vaccine showed that ELISA measured maximal titres of primary response at 23 days post-vaccination (dpv), and at day 17 of secondary response, while PH detected maximal titres for primary as well as secondary response around day 60 pv. Mice immunized with OE vaccine studied by ELISA presented a plateau of primary response around 40 dpv, while secondary response was maximal around 80 dpv. The same sera tested by PH showed the highest titres for primary response on day 50 pv and secondary response was maximal on day 40 pv. A criterion for the evaluation of vaccination efficiency in the murine model is proposed based on the methods employed in the determination of antibody level.

Comparación de distintos métodos para la medición de la respuesta inmune humoral de ratones vacunados con el virus de la fiebre aftosa. / [Comparison of various methods for the measurement of the humoral immune response in mice vaccinated with aphthous fever virus]

Kinetics of the humoral immune primary response and seven-day secondary response of adult CF1 mice to FMDV O1 Campos adjuvanted in aluminium hydroxide-saponin (AHS) or in oil emulsion (OE) were evaluated by means of ELISA and passive hemagglutination (PH). Analysis of the response to AHS vaccine showed that ELISA measured maximal titres of primary response at 23 days post-vaccination (dpv), and at day 17 of secondary response, while PH detected maximal titres for primary as well as secondary response around day 60 pv. Mice immunized with OE vaccine studied by ELISA presented a plateau of primary response around 40 dpv, while secondary response was maximal around 80 dpv. The same sera tested by PH showed the highest titres for primary response on day 50 pv and secondary response was maximal on day 40 pv. A criterion for the evaluation of vaccination efficiency in the murine model is proposed based on the methods employed in the determination of antibody level.

Protection conferred by TrpE fusion proteins containing portions of the C-terminal region of capsid protein VP1 of foot-and-mouth disease virus.

Major immunogenic sites of foot-and-mouth disease virus (FMDV) have been mapped to the C-terminal third of capsid protein VP1; we studied the immunogenicity of a series of TrpE-FMDV fusion proteins containing this region of FMDV strain O1 Campos. Fusion protein TrpE-dCN, which contains a dimer of VP1 amino acid sequences consisting of amino acids 200 to 213 linked by a diproline spacer to amino acids 141 to 158 (200-213 approximately P-P-G approximately 141-158), induced the best response. A single inoculation of guinea-pigs with 100 micrograms TrpE-dCN elicited high levels of neutralizing antibodies and protected all the animals against challenge infection with homologous virus. Although the closely related FMDV strains O1 Campos and O1 Caseros induced high levels of cross-protection, TrpE-dCN-vaccinated guinea-pigs were poorly protected against challenge infection with heterologous FMDV strain O1 Caseros. Nucleotide sequence analysis revealed that amino acid differences at residues 149 and 152 were critical for the induction of cross-protection and that neutralizing epitopes not present in TrpE-dCN are likely to be responsible for conferring a high level of cross-protection between FMDV strains O1 Campos and O1 Caseros.

[Immune response in mice induced by different doses of foot-and-mouth disease virus]. / Respuesta inmune en ratones inducida por diferentes dosis del virus de la Fiebre Aftosa.

Response against foot-and-mouth disease virus 01 Campos on vaccinated female mice and suckling litter was estimated. Vaccine doses ranged between 0.05 and 30 micrograms of virus. The lower dose protected 39.5% of suckling litter whereas 100% protection was achieved using the higher doses. Serum neutralization indexes in females, at 45 days post vaccination, ranged from 3.60 to 4.86 for 0.05 microgram to 0.10 microgram and 4.86 to 5.50 for the high doses. Although the doses were excessively high (eq. 1 mu/g average live weight) and (OH)3Al was used as adjuvant, no adverse effect on the immune system was detected in any of the vaccinated animals.

Respuesta inmune en ratones inducida por diferentes dosis del virus de la Fiebre Aftosa. / [Immune response in mice induced by different doses of foot-and-mouth disease virus]

Response against foot-and-mouth disease virus 01 Campos on vaccinated female mice and suckling litter was estimated. Vaccine doses ranged between 0.05 and 30 micrograms of virus. The lower dose protected 39.5

of suckling litter whereas 100

protection was achieved using the higher doses. Serum neutralization indexes in females, at 45 days post vaccination, ranged from 3.60 to 4.86 for 0.05 microgram to 0.10 microgram and 4.86 to 5.50 for the high doses. Although the doses were excessively high (eq. 1 mu/g average live weight) and (OH)3Al was used as adjuvant, no adverse effect on the immune system was detected in any of the vaccinated animals.

[Use of sarcoma 180 TG cells for obtaining ascitic fluid from mice hyperimmune to the foot-and-mouth disease virus]. / Uso de las células de sarcoma 180 TG para la obtención de fluídos ascíticos hiperinmunes de ratón para virus aftoso.

Mice immunized with FMDV C3 Arg 84 antigen were inoculated intraperitoneally with sarcoma 180 TG. The ascitic fluid obtained by ventral puncture contained high titers of antibodies, similar to those obtained from serum, as determined by neutralization and ELISA tests. Ascitic volumes were 10 to 20 times greater than those obtainable with.

Uso de las células de sarcoma 180 TG para la obtención de fluídos ascíticos hiperinmunes de ratón para virus aftoso. / [Use of sarcoma 180 TG cells for obtaining ascitic fluid from mice hyperimmune to the foot-and-mouth disease virus]

Mice immunized with FMDV C3 Arg 84 antigen were inoculated intraperitoneally with sarcoma 180 TG. The ascitic fluid obtained by ventral puncture contained high titers of antibodies, similar to those obtained from serum, as determined by neutralization and ELISA tests. Ascitic volumes were 10 to 20 times greater than those obtainable with.

[Neutralization of foot-and-mouth disease virus O1 Campos by antibodies induced by a synthetic peptide]. / Neutralización del virus de la fiebre aftosa O1 Campos por anticuerpos inducidos por un péptido sintético.

Foot-and-Mouth disease virus (FMDV), contains a positive single-stranded RNA enclosed in a protein capsid. Previous studies have shown that a synthetic peptide located at the carboxyterminal end of VP1 of FMDV strain O1 Kaufbeuren (O1K) at the amino acid positions 144-159, and coupled to KLH was able to elicit high titers of neutralizing antibodies in guinea pigs and protected them against challenge with the homologous virus (8, 15). FMDV strain O1 Campos (O1 Ca) has a similar amino acid sequence to O1K, differing in amino acid 144 (leucin instead valine). We report that the O1K synthetic peptide 144-159 protects adult mice against challenge with FMDV O1Ca when the peptide is coupled to KLH with glutaraldehyde as coupling agent (Table 2) whereas protection is very low when other carrier proteins are used.

Neutralización del virus de la fiebre aftosa O1 Campos por anticuerpos inducidos por un péptido sintético / Neutraliztion of fost-and-mouth diseases virus O1 Campos by antibodies induced by synthetic peptide

Un hexadecapéptido correspondiente a la secuencia 144-159 de la proteína estructural VP1 del virus de la fiebre aftosa (CFA), cepa O1K, fue sintetizado por métodos químicos. El mismo, acoplado covalentemente a distintas proteínas portadoras, demostró su capacidad de inducir anticuerpos neutralizantes del VFA cepa O1 Campos (O1Ca) en ratones adultos. La protección fue del 100% cuando se lo acopló a la hemocianina de un molusco (KLH) y fue inoculado en cantidad de 60 microng, dividida en dos dosis y adyuvada con mezcla oleosa. Las proteínas utilizadas como portadoras: albúmina bovina, purificado proteico de Mycobacterium (PPD), gliadina y células de Brocelas produjeron una respuesta inmune menor que la KLH

Neutralización del virus de la fiebre aftosa O1 Campos por anticuerpos inducidos por un péptido sintético. / [Neutralization of foot-and-mouth disease virus O1 Campos by antibodies induced by a synthetic peptide]

Foot-and-Mouth disease virus (FMDV), contains a positive single-stranded RNA enclosed in a protein capsid. Previous studies have shown that a synthetic peptide located at the carboxyterminal end of VP1 of FMDV strain O1 Kaufbeuren (O1K) at the amino acid positions 144-159, and coupled to KLH was able to elicit high titers of neutralizing antibodies in guinea pigs and protected them against challenge with the homologous virus (8, 15). FMDV strain O1 Campos (O1 Ca) has a similar amino acid sequence to O1K, differing in amino acid 144 (leucin instead valine). We report that the O1K synthetic peptide 144-159 protects adult mice against challenge with FMDV O1Ca when the peptide is coupled to KLH with glutaraldehyde as coupling agent (Table 2) whereas protection is very low when other carrier proteins are used.

Neutralización del virus de la fiebre aftosa O1 Campos por anticuerpos inducidos por un péptido sintético / Neutraliztion of fost-and-mouth diseases virus O1 Campos by antibodies induced by synthetic peptide

Un hexadecapéptido correspondiente a la secuencia 144-159 de la proteína estructural VP1 del virus de la fiebre aftosa (CFA), cepa O1K, fue sintetizado por métodos químicos. El mismo, acoplado covalentemente a distintas proteínas portadoras, demostró su capacidad de inducir anticuerpos neutralizantes del VFA cepa O1 Campos (O1Ca) en ratones adultos. La protección fue del 100% cuando se lo acopló a la hemocianina de un molusco (KLH) y fue inoculado en cantidad de 60 microng, dividida en dos dosis y adyuvada con mezcla oleosa. Las proteínas utilizadas como portadoras: albúmina bovina, purificado proteico de Mycobacterium (PPD), gliadina y células de Brocelas produjeron una respuesta inmune menor que la KLH (AU)