ESTUDIO DE LA SENSIBILIZACIÓN POR AEROALÉRGENOS CON ESPECIAL ENFOQUE EN LA POLINOSIS COMO FACTOR DE SENSIBILIZACIÓN / Sensitizen by aeroalergens study with special focus on polinosis as sensitizen factor

Fundamento. Las alergias son importantes en Salud Pública; entre distintos tipos, la sensibilidad polínica (SP), en su forma de hipersensibilidad a pólenes, son patologías que afectan a parte de la población. Objetivo. Valorar la importancia del problema de SP en la región y relacionarlas con rinitis, asma, valores de IgE y otras alergias. Lugar de aplicación. Servicio Cátedra de Alergia e Inmunología (SAEI) del Hospital Nacional de Clínicas (HNC), Región Centro del País. Diseño. Estudio de corte. Estudio descriptivo útil para el administrador sanitario ya que permite determinar la carga que significa la enfermedad en la población. Permite conocer la prevalencia (P) de enfermedad. Población. Se analizaron historias clínicas (HC) de los pacientes que concurrieron al SAEI, ubicado en Córdoba capital. Método. Se procesaron las HC de los pacientes que acudieron al SAEI durante los años 2008-09 y a los que se les realizó el prick test, usando el programa Epi-Info 2000 versión 3.3.2. Se las analizó estadísticamente determinando Chicuadrado e intervalo de confianza. Resultados. La P de SP fue de 51,96%. Del total de la muestra, el 24,24% era monosensibilizado el 12,50% de estos lo era a pólenes. Del 75,76% de la muestra restante (polisensibilizado), el 65,33% estaba sensibilizado a pólenes. Conclusión. Analizando la P obtenida se desprende que las SP constituyen una problemática que afecta a más de la mitad de la población estudiada. También se observó que dicha problemática tiene, para la mayoría de los pacientes, una comorbilidad asociada a otros tipos de alergia. (AU)

Background. Allergies are important in Public Health, among the different types Pollen sensitivity, as hypersensitivity to pollen, are pathologies affecting part of the population. Objective. To appreciate the importance of the Pollen sensitivity problem in our region and to relate them to Rhinitis, Asthma, IgE values, and other allergies. Setting. Allergy and Immunology Service (AAIS) of the Hospital Nacional de Clínicas (HNC), in Córdoba city, Central Region of the country. Design. A Cross-sectional study was developed. This is a descriptive study of usefulness for the healthcare administrator because it allows estimating the disease burden significance on the population. It lets to know the sickness Prevalence (P). Population. Clinical Histories (CH) of the patients that attended the AAIS, located in Córdoba city were analyzed. Methods. CH of the patients that attended the AAIS during the years 2008 and 2009 and that the Prick Test was carried out, were processed by using the 2000 Epi-Info Program, 3.3.2. version. Statistically, Chi-square and Confidence Interval were calculated. Results. The Prevalence of Pollen sensitivity was 51.96%. From the total sample, 24.24% was monosensitized patients. Among them, 12.5% were monosensitized to pollens. The rest of the sample (75.76%) were poly-sensitized, from these, 65.33% were sensitized to pollens. Conclusion. Analyzing the obtained Prevalence emerges that Pollen sensitivity represent a problematic that affect more than a half of the studied population. Also it was observed that the pathology presented a comorbidity associated to other types of allergy produced by mites or fungus, among others, for the majority of the patients.(AU)

The coat protein of tobamovirus acts as elicitor of both L2 and L4 gene-mediated resistance in Capsicum.

In Capsicum, the resistance conferred by the L(2) gene is effective against all of the pepper-infecting tobamoviruses except Pepper mild mottle virus (PMMoV), whereas that conferred by the L(4) gene is effective against them all. These resistances are expressed by a hypersensitive response, manifested through the formation of necrotic local lesions (NLLs) at the primary site of infection. The Capsicum L(2) gene confers resistance to Paprika mild mottle virus (PaMMV), while the L(4) gene is effective against both PaMMV and PMMoV. The PaMMV and PMMoV coat proteins (CPs) were expressed in Capsicum frutescens (L(2)L(2)) and Capsicum chacoense (L(4)L(4)) plants using the heterologous Potato virus X (PVX)-based expression system. In C. frutescens (L(2)L(2)) plants, the chimeric PVX virus containing the PaMMV CP was localized in the inoculated leaves and produced NLLs, whereas the chimeric PVX containing the PMMoV CP infected the plants systemically. Thus, the data indicated that the PaMMV CP is the only tobamovirus factor required for the induction of the host response mediated by the Capsicum L(2) resistance gene. In C. chacoense (L(4)L(4)) plants, both chimeric viruses were localized to the inoculated leaves and produced NLLs, indicating that either PaMMV or PMMoV CPs are required to elicit the L(4) gene-mediated host response. In addition, transient expression of PaMMV CP into C. frutescens (L(2)L(2)) leaves and PMMoV CP into C. chacoense (L(4)L(4)) leaves by biolistic co-bombardment with a beta-glucuronidase reporter gene led to the induction of cell death and the expression of host defence genes in both hosts. Thus, the tobamovirus CP is the elicitor of the Capsicum L(2) and L(4) gene-mediated hypersensitive response.

Biological and molecular characterization of P101 isolate, a tobamoviral pepper strain from Bulgaria.

A tobamovirus isolated from pepper crops in Bulgaria has been characterized, and is referred to below as P101. It was closely related to Paprika mild mottle virus (PaMMV) (Dutch isolate), based upon the serological relationship of its coat protein, and the nucleotide sequence analysis of the gene encoding the coat protein and the 3' non-coding region of the viral RNA. The coat proteins of the two isolates differ by two amino acids, and these substitutions may be responsible for the different reactivity of the isolates towards a polyclonal antiserum raised against the virion of the Dutch isolate. The biological behaviour of both isolates was similar in the hosts tested, except in pepper plants where P101 induced delayed and milder symptoms compared with PaMMV, although their accumulation levels were similar. In addition, we investigated the infection pattern of the two isolates in tomato plants. Both isolates accumulated in protoplasts as well as in inoculated leaves, although systemic invasion was limited. This limited spread was not due to activation of defense mechanism(s) in the plant, since the upper uninoculated leaves from P101-infected tomato plants were fully susceptible to challenge inoculation with the virus. Instead, it appears due to a restriction of long-distance movement, that could be overcome in tomato plants co-infected with Tobacco mosaic virus (TMV), but not with either Cucumber mosaic virus or Pepino mosaic virus. The ability of P101 to move systemically in the presence of TMV was not linked to enhanced accumulation of P101 at the cellular level. Thus, a tobamovirus but not the viruses tested from other genera could complement, in trans, the function(s) required for PaMMV to invade the upper uninoculated leaves. Paprika mild mottle virus strain B is proposed as the name for this new isolate.

[Effectiveness and impact in the quality of life of ketotifen ophthalmic solution. Results of zeta study in patients with seasonal allergic conjunctivitis]. / Efectividad e impacto en la calidad de vida del colirio de ketotifeno. Resultados del estudio zeta en pacientes con conjuntivitis alérgica estacional.

PURPOSE: To study the effectiveness of ketotifen ophthalmic solution (0.25 mg/ml) in seasonal allergic conjunctivitis (SAC) and the impact on the patient's quality of life. METHODS: A multicentric, longitudinal, prospective study was designed. 284 Spanish ophthalmologists participated recruiting 1145 patients with SAC. After obtaining the informed consent, a drop of ketotifen ophthalmic solution was instilled. At the visit, clinical symptoms pre and post-treatment were assessed. The patients answered a questionnaire of quality of life (QOL) pre-treatment and minimum one week after initiating the treatment. The qualitative variables were described by the percentage, and the quantitative were described by the average, median, standard deviation, and maximum and minimum values. The effectiveness (change of intensity of the symptoms) and the quality of life were studied by the Wilcoxon test with a significance level of 5% (alpha = 0.05). RESULTS: Following the instillation of the ketotifen ophthalmic solution the intensity of the ocular symptoms (redness, edema, tearing, secretion, photophobia and visual acuity impairment) decreased significantly. Comparing both QOL, we observed a statistically significant reduction of the limitation perceived by the patients in their daily activities, animic state and ocular symptoms. In 0,7% some adverse event was referred, none was serious and only in one case the probable relationship with the drug was specified. CONCLUSION: The results of the ZETA study demonstrate the tolerability and effectiveness of the ketotifen ophthalmic solution for all the symptoms of SAC in clinical practice, observing improvement in the quality of life of the patient.

Altered local and systemic spread of movement deficient virus in transgenic tobacco plants expressing the cucumber mosaic virus 3a protein.

The 3a protein encoded by RNA 3 of cucumber mosaic virus has been identified as the viral cell-to-cell movement protein. The constitutive expression in transgenic tobacco plants of 3a protein from a subgroup I strain was able to complement in trans the short distance movement of a 3a defective CMV mutant belonging to a different taxonomic subgroup. This ability was dependent upon the accumulation levels of the 3a protein in transgenic tobacco plants. However, an initial delay in viral accumulation and spread of the defective virus as compared to the wild type virus was determined in complementation tests. Furthermore, a reduction in disease symptoms as well as a different pattern of systemic viral distribution from those of the wild type virus was detected. These results show that the early events in viral infection affect the long distance spread of the virus. Finally, the wild type virus moved faster in the 3a protein-expressing plants than in control plants, thus indicating that the constitutive expression of the 3a protein favours long-distance viral spread.

The coat protein is required for the elicitation of the Capsicum L2 gene-mediated resistance against the tobamoviruses.

In Capsicum, the resistance against tobamoviruses conferred by the L2 gene is effective against all but one of the known tobamoviruses. Pepper mild mottle virus (PMMoV) is the only virus which escapes its action. To identify the viral factors affecting induction of the hypersensitive reaction (HR) mediated by the Capsicum spp. L2 resistance gene, we have constructed chimeric viral genomes between paprika mild mottle virus (PaMMV) (a virus able to induce the HR) and PMMoV. A hybrid virus with the PaMMV coat protein gene substituted in the PMMoV-S sequences was able to elicit the HR in Capsicum frutescens (L2L2) plants. These data indicate that the sequences that affect induction of the HR mediated by the L2 resistance gene reside in the coat protein gene. Furthermore, a mutant that codes for a truncated coat protein was able to systemically spread in these plants. Thus, the elicitation of the host response requires the coat protein and not the RNA.

Attachment of Agrobacterium tumefaciens B6 and A. radiobacter K84 to Tomato Root Tips.

Agrobacterium tumefaciens B6 and the avirulent Agrobacterium radiobacter strain K84 attached to in vitro-cultured tomato root tips, but the binding of strain B6 to root tips was greater than the binding of strain K84. Strain K84 was not able to block the attachment of A. tumefaciens B6 to in vitro-cultured tomato root tips.

Resistance to pepper mild mottle tobamovirus conferred by the 54-kDa gene sequence in transgenic plants does not require expression of the wild-type 54-kDa protein.

We previously reported that Nicotiana benthamiana plants transformed with the wild-type 54-kDa region of the pepper mild mottle tobamovirus, S strain (PMMoV-S), displayed two different resistance responses against PMMoV infection. Some of the transgenic plants exhibited a complete and highly resistant phenotype while the remaining plants showed a delayed resistance (Tenllado et al., 1995, Virology 211, 170--183). Here we show that some of the N. benthamiana plants transformed with a construct expressing a PMMoV-S truncated 54-kDa protein coding sequence also displayed a complete and highly resistant phenotype similar to that shown by the wild-type 54-kDa transgenic plants. This result indicates that the wild-type, full-length 54-kDa protein is not required in mediating the complete resistance phenotype against PMMoV. The remaining truncated 54-kDa transgenic plants were susceptible to PMMoV infection but showed a variable delay in the appearance of symptoms. Unlike the wild-type 54-kDa transgenic plants, which were initially susceptible to the infection but recovered later, the truncated 54-kDa transgenic plants never exhibited this delayed resistance phenotype. However, they displayed a new type of altered symptomatic phenotype. The truncated 54-kDa transgenic lines also exhibited a lower level of transgenic transcripts compared to the wild-type 54-kDa transgenic lines which could account for the absence of the delayed resistance phenotype.

Accumulation kinetics of CMV RNA 3-encoded proteins and subcellular localization of the 3a protein in infected and transgenic tobacco plants.

The complete nucleotide sequence of RNA 3 of a Spanish isolate of cucumber mosaic virus (CMV-24) has been determined. The encoded putative cell-to-cell movement protein (3a protein) and the coat protein are 279 and 218 amino acids long, respectively. The 3a protein was expressed in Escherichia coli using the vector pT7-7 and was used to raise an immunoserum. We have followed the time course of accumulation of the 3a protein, in parallel to that of the coat protein, and its subcellular localization as a function of time after CMV-24 infection on tobacco plants. The maximum accumulation level of the 3a protein was reached at early stages of infection, being detected in the cytosolic and the cell wall fractions. At later stages of infection, a decline in accumulation levels of the 3a protein was observed, and the protein was essentially associated with the cell wall fractions. These data were corroborated by immunocytochemistry performed in both infected and 3a-expressing transgenic tobacco plants.

Nicotiana benthamiana plants transformed with the 54-kDa region of the pepper mild mottle tobamovirus replicase gene exhibit two types of resistance responses against viral infection.

Nicotiana benthamiana plants transformed with the 54-kDa region of the pepper mild mottle tobamovirus (PMMV) replicase gene were generated and six independently transformed plant lines were analyzed for resistance to PMMV. Two different resistance responses were obtained. Some of the transgenic plants from only two lines showed a preestablished, complete, and highly resistant phenotype since no viral symptoms were observed, although a low level of virus replication occurred. The remaining plants from these two lines and all of the plants from the other four lines tested showed a delayed, induced, and also highly resistant phenotype since they were susceptible early, but were able to recover from the systemic PMMV infection. Recovered, symptomless leaves were resistant to the PMMV strain from which the 54-kDa gene was derived and to a closely related strain but not to tobacco mosaic virus. Such a delayed resistance phenotype has not been previously described for any plant expressing viral replicase sequences. The transgenic plants within the lines displaying complete or delayed resistance phenotypes were analyzed for transgene expression before and after PMMV inoculation and the two types of resistance responses were shown to be independent of the transgene transcript level.

The Capsicum L3 gene-mediated resistance against the tobamoviruses is elicited by the coat protein.

The L3 gene is responsible for the hypersensitive response in Capsicum plants against infection by tobamoviruses. The resistance conferred by this gene is one of the most effective so far described against tobamoviruses. Certain isolates of pepper mild mottle virus (PMMV) are the only tobamoviruses able to overcome the L3 resistance. Chimeric viral genomes between PMMV-S (to which L3 plants are hypersensitive) and PMMV-I (an L3 resistance-breaking isolate) led us to conclude that sequence variation within the coat protein gene of both isolates determines their different virulence in L3L3 plants. Furthermore, the results indicate that a single amino acid substitution, Asn to Met, at position 138 of the PMMV-I coat protein is sufficient to induce the hypersensitive response and localization of viral infection in C. chinense plants. Finally, the use of a mutant coding for a truncated coat protein (maintaining the Met138 coding sequence at the RNA level) demonstrates that a functional coat protein is required for elicitation of the L3 gene-mediated resistance.

The 3a protein from cucumber mosaic virus increases the gating capacity of plasmodesmata in transgenic tobacco plants.

The 3a protein, encoded by RNA 3 of cucumber mosaic virus (CMV), is the putative movement protein of viral progeny in infected plants. An analysis of transgenic tobacco plants constitutively expressing the CMV 3a protein showed that the protein is accumulated in leaves at every stage of development. In fully expanded leaves the protein is immunodetectable mostly in a cell-wall-enriched fraction. Dye-coupling experiments using fluorescent-dextran probes were performed on fully expanded leaves to study the modifying effect of CMV 3a protein on the gating capacity of plasmodesmata. Movement of fluorescein-isothiocyanate-labelled dextran with a mean molecular mass of 10,000 Da, and an approximate Stokes' radius of 2.3 nm, was detected between cells of the 3a protein transgenic plants, but not in the control plants. These results are consistent with the idea that the CMV 3a protein is involved in the modification of plasmodesmata and, therefore, in the cell-to-cell spread of the virus.

Rapid detection and differentiation of tobamoviruses infecting L-resistant genotypes of pepper by RT-PCR and restriction analysis.

A procedure involving reverse transcription followed by polymerase chain reaction (RT-PCR) was developed for typing pathotypes of the tobamoviruses infecting the L-resistant genotypes of pepper. The method provides a much simpler alternative to the bioassay tests for the different Capsicum spp. genotypes previously used. Discrimination between the two pathotypes, P1,2 and P1,2,3, which cannot be differentiated by serological means, was achieved by restriction enzyme analysis of the PCR products. The assay also detects and distinguishes both pathotypes in a single mixed-infected plant. The procedure should be useful for the diagnosis and control of the disease and helpful to breeders and biotechnologists when producing and evaluating resistance in pepper plants.

The nucleotide sequence of the coat protein genes and 3' non-coding regions of two resistance-breaking tobamoviruses in pepper shows that they are different viruses.

The nucleotide sequence of the coat protein genes and 3' non-coding regions of two different resistance-breaking tobamoviruses in pepper have been determined. The deduced coat protein of an Italian isolate of pepper mild mottle virus (PMMV-I) consists of 156 amino acids and its 3' non-coding region is 198 nucleotides long. They have been found to be very similar in sequence and structure to those previously reported for a Spanish isolate (PMMV-S). In contrast, a Dutch isolate termed P 11 codes for a coat protein of 160 amino acids and its 3' non-coding region is 291 nucleotides long, which may have arisen by duplication. The nucleotide and the predicted coat protein amino acid sequence analysis show that this isolate should be considered as a new virus within the tobamovirus group. The term paprika mild mottle virus (PaMMV) is proposed.

Nucleotide sequence of the genomic RNA of pepper mild mottle virus, a resistance-breaking tobamovirus in pepper.

The entire genomic RNA of a Spanish isolate of pepper mild mottle virus (PMMV-S), a resistance-breaking virus in pepper, was cloned and sequenced and shown to be similar to other tobamoviruses in its genomic organization. It consisted of 6357 nucleotides (nt) and contained four open reading frames (ORFs) which encode a 126K protein and a readthrough 183K protein (nt 70 to 4908), a 28K protein (nt 4909 to 5682) and a 17.5K coat protein (nt 5685 to 6158). This is the first tobamovirus in which none of the ORFs overlap. Both its nucleic acid and predicted protein sequences were compared with the previously determined sequences of other tobamoviruses. The variations and similarities found and their relationship with the pathogenicity of this virus are discussed.

Metabolism of halothane in obese Fischer 344 rats.

Halothane is metabolized by an oxidative pathway to stable, nonvolatile end products, trifluoroacetic acid (TFAA) and bromide (Br-), and by reductive pathways to Br-and inorganic fluoride (F-). There is evidence that both oxidatively and reductively formed intermediates may produce hepatotoxicity, although the exact etiology of the fulminant hepatic necrosis seen in humans is unproven. Obese patients receiving volatile anesthetics exhibit higher serum anesthetic metabolite concentrations than do normal-weight patients, and thus might be at greater risk of hepatotoxicity because of higher concentrations of reactive intermediates from halothane metabolism. To eliminate the variables inherent in human clinical studies leading to confounding interpretation of data, this study determined the contributions of oxidative and reductive pathways to halothane metabolism in an animal model of human hypertrophic obesity, the most common form of human obesity. Eight pairs of obese (high-fat diet) and normal-weight (standard chow), male Fischer 344 rats were anesthetized with halothane for 4 h at an inspired concentration of 0.78%. Serum and urinary concentrations of TFAA, Br-, and F-were measured. Thirty-six hours following halothane anesthesia, mean serum TFAA concentrations peaked at 7.3 +/- 1.1 mM in obese rats and 4.7 +/- 0.7 mM in nonobese rats. TFAA urinary excretions during the 180-h period postanesthesia were 519 +/- 69 and 336 +/- 22 mumol, respectively. Peak serum Br- concentrations were 9.1 +/- 1.0 and 6.9 +/- 0.6 mM for obese and nonobese rats, respectively, and Br-urinary excretions were 127 +/- 30 and 79 +/- 14 mumol, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Hemagglutinating activity in phytopathogenic bacteria surface compounds.

Extracellular components of plant pathogenic bacteria were obtained from their culture medium as well as from the whole cells by using NaCl 1 M, pH 6.0; 20% sucrose dissolved in 0.03 M Tris buffer, pH 8.0; or 0.05 M Na2EDTA. All the extracts from Erwinia carotovora subsp. carotovora, Xanthomonas campestris pv. campestris, Pseudomonas syringae pv. phaseolicola, Xanthomonas campestris pv. phaseoli, Pseudomonas solanacearum, and Erwinia carotovora subsp. atroseptica, were assayed for hemagglutinating activity on sheep, rabbit and chicken red blood cells (RBCs). The only active extracts were those obtained by NaCl treatment. They agglutinated sheep and rabbit erythrocytes. Extracts from E. carotovora subsp. atroseptica gave rise to the high agglutination titer on rabbit RBCs. These extracts had the lowest polysaccharide/protein ratio. E. carotovora subsp. carotovora extracts showed only a low titer (18.5 units). The agglutinating activity present in NaCl extracts of the bacteria tested was inhibited by different carbohydrates to various extent. Extracts from E. carotovora subsp. atroseptica appeared to be the most sensitive ones while those of E. carotovora subsp. carotovora least sensitive to the presence of sugar. It is suggested that hemagglutinins observed in plant pathogenic bacteria and those in plant host are similar and that both may, in some way, be involved in the plant-parasite relationship.

Purification and identification of rat hepatic cytosolic enzymes responsible for defluorination of methoxyflurane and fluoroacetate.

Enzymes responsible for the defluorination of methoxyflurane (MOF) and fluoroacetate (FAc) were separated and purified from rat liver cytosol. Both hepatic cytosolic enzymes with defluorination activity were labile and addition of 2-mercaptoethanol had little effect on the stability of these enzymes. Glutathione S-transferase (GT) activity of the same cytosolic fractions was stable for at least 11 days. Separation of defluorination and GT enzymatic activities on DEAE-Sephadex A-50 and reduced glutathione-affinity columns revealed that the defluorinations of MOF and FAc were primarily catalyzed by anionic proteins which also exhibited GT activity. Further identification by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed protein bands with pl values of approximately 6.5 and 6.9 and molecular weights of approximately 20,000. However, other proteins that exhibited no GT activity also defluorinated MOF and FAc, but accounted for only 10% of the total defluorination activity present in anionic proteins. Results from a separate purification experiment using a CM-cellulose column also indicated that the enzymes responsible for defluorination coeluted with cationic GTs. Collectively, these cationic enzymes were responsible for about 20% of the recovered cytosolic defluorination activities. The results suggest that the cytosolic defluorinations of both MOF and FAc are primarily the result of a dehalogenation reaction catalyzed by one or more species of rat liver cytosolic GTs.