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Summary: Hereditary angioedema (HAE) poses a high burden of disease, being its epidemiological and clinical data heterogeneous among countries, with no recent published studies concerning Portuguese patients. Therefore, we aimed to raise awareness of HAE and to contribute to clinical knowledge. An observational, descriptive, retrospective, and cross-sectional study was performed, that included a cohort of 126 patients followed in a single Portuguese Center. We observed a high prevalence of HAE-C1-INH type II (45.2% of patients). Most HAE patients (67.4%) presented the initial manifestations of the disease before adulthood, at a mean age of 12.6 ± 8.4 years. However, we found a long delay in HAE diagnosis, especially in those without family history (mean 20.7 ± 17.3 years). Stress was the most common trigger, followed by trauma and infection. Symptoms involving different systems were increasingly reported with increased disease duration. Cutaneous symptoms (95.0%) were more frequent, followed by gastrointestinal (80.7%), and respiratory symptoms (50.4%). HAE symptoms led to abdominal surgery in 22 (17.5%) patients and induced laryngeal edema requiring intubation/tracheostomy in 8 (6.3%) patients. Most patients were under long-term prophylaxis, mainly with attenuated androgens (62.7% of patients).The correct distinction between HAE and other common causes of angioedema is critical, allowing reduction of diagnostic delay, improvement of adequate management, and ultimately improving outcomes and quality of life of HAE patients.
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BACKGROUND: Autoimmune chronic spontaneous urticaria (CSU) is due to mast cell (MC)-activating autoantibodies, which are screened for by the autologous serum skin test (ASST) and basophil tests (BTs). Many CSU patients are positive in only one of these tests. How often this occurs and why is currently unknown. OBJECTIVES: To characterize the prevalence of mismatched ASST and BTs in CSU patients, and to investigate possible reasons for these mismatches. METHODS: We determined the rates of ASST+/BT- and ASST-/BT+ mismatches in published CSU studies. We assessed sera from 48 CSU patients by ASST, two BTs (basophil histamine release assay, BHRA; basophil activation test, BAT), a MC histamine release assay (MCHRA) and by ex vivo skin microdialysis (SMD). RESULTS: The ASST/BT mismatch rate in published CSU studies was 31% (ASST+/BT-: 22%, ASST-/BT+: 9%). In our patients, the ASST/BHRA and ASST/BAT mismatch rate was 35.4% (ASST+/BHRA-: 18.8% and ASST-/BHRA+: 16.7%) and 31.3% (ASST+/BAT-: 6.3% and ASST-/BAT+: 25.0%), respectively, and the two BTs were significantly correlated (P = 0.0002). The use of heterologous MCs, in vitro and in situ, instead of basophils produced similar results (MCHRA mismatch: 47.9%, ASST+/MCHRA-: 18.8%, ASST-/MCHRA+: 29.2%; SMD mismatch: 40.0%, ASST+/SMD-: 10.0% and ASST-/SMD+: 30.0%), and the MCHRA was highly correlated with SMD results (P = 0.0002). CONCLUSIONS: The ASST and BTs show divergent results in a third of CSU patients. Mismatches cannot be explained by the choice of basophil assay, the type of heterologous cells exposed to CSU serum in vitro (basophils vs. mast cells), nor the experimental setting of heterologous skin mast cells (in vitro vs. in situ). Thus, serum-induced whealing, in CSU patients, seems to involve autologous skin signals modulating MC degranulation.
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Urticaria Crónica , Urticaria , Basófilos , Enfermedad Crónica , Humanos , Pruebas Cutáneas , Urticaria/diagnósticoRESUMEN
BACKGROUND AND OBJECTIVES: The prevalence of fish allergy has increased in recent years. The parvalbumin Gad c 1 is a major cod allergen that is used as a follow-up marker in patients with fish allergy. Objectives: To determine the clinical and laboratory characteristics of a population of patients with fish allergy. To analyze the role of the specific IgE (sIgE) of recombinant Gad c 1 (rGad c 1) and skin prick tests (SPTs) in confirming the acquisition of tolerance to fish. METHODS: We performed a retrospective study of patients with fish allergy from July 1, 2005 to December 31, 2016. The population was characterized according to demographic data, species of fish associated with allergic reactions, and symptoms. The SPT wheal diameter and sIgE for fish and rGad c 1 were evaluated before acquisition of tolerance (T0) and afterwards (T1). RESULTS: The study population comprised 81 patients (68% male). Most reactions were triggered by hake (51%), mackerel (30%), and cod (26%). The most frequent manifestations were urticaria/angioedema (72%), gastrointestinal symptoms (35%), and eczema (33%); 42% of patients experienced anaphylaxis. At T0, the average sIgE values were as follows: cod, 32.2 kUA/L; sardine, 18.4 kUA/L; hake, 17.5 kUA/L; salmon, 13.9 kUA/L; tuna, 4.5 kUA/L; and rGad c 1, 22.9 kUA/L. In patients who acquired tolerance to at least 1 fish species (n=60; 74%), the mean value of rGad c 1 at T1 (5.1 kUA/L) was significantly lower than at T0 (16.8 kUA/L) (P=.001). Significant values were also recorded for the average diameter of the SPT wheal and the evaluations at T0 and T1 for hake (9.42 mm/3.79 mm) and salmon (7.8 mm/2.8 mm) (P=.002 and P=.026, respectively). CONCLUSION: The decrease in sIgE to rGad c 1 and the mean wheal diameter of SPT for hake and salmon can be used as markers of prognosis in the acquisition of tolerance by fish-allergic patients.
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Alérgenos/inmunología , Peces/inmunología , Hipersensibilidad a los Alimentos/diagnóstico , Hipersensibilidad a los Alimentos/inmunología , Parvalbúminas/inmunología , Proteínas Recombinantes/inmunología , Adolescente , Adulto , Animales , Femenino , Humanos , Inmunoglobulina E/inmunología , Masculino , Pronóstico , Estudios Retrospectivos , Pruebas Cutáneas , Adulto JovenRESUMEN
Summary: Objective. Describe the safety and long-term use of omalizumab in chronic urticaria (CU), both spontaneous (CSU) and inducible (CIndU). Methods. Retrospective chart-review (2006-15) of CU patients treated with omalizumab for ≥ 6 months. Statistical analyses: descriptive statistics, Mann-Whitney, generalized linear models. Results. 23 patients with CSU (3 men), 3 with CIndU (2 men). Generalized linear models showed UAS reduction per omalizumab administration of 16% in CIndU and CSU and UAS7, of 15% in CIndU, and 20% in CSU. DLQI score at baseline had a median of 19 (CIndU and CSU) and after omalizumab a median of 0 (in both). Seven CSU patients stopped omalizumab and remain asymptomatic. No side-effects were observed. Conclusion. Omalizumab is safe and efficacious in CU. Stopping omalizumab can be tried, as some patients achieve remission.
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Antialérgicos/uso terapéutico , Omalizumab/uso terapéutico , Urticaria/tratamiento farmacológico , Adulto , Antialérgicos/efectos adversos , Femenino , Humanos , Inmunoglobulina E/sangre , Masculino , Persona de Mediana Edad , Omalizumab/efectos adversos , Portugal , Estudios Retrospectivos , Resultado del Tratamiento , Adulto JovenRESUMEN
SUMMARY: Eosinophilic esophagitis (EoE) is an increasingly frequent diagnosis in our clinical practice, mainly in pediatric age. Allergic responses to food and aeroallergens have been increasingly implicated in the etiology of this disease. We describe a retrospective data analysis of pediatric EoE patients followed in our Immunoallergology Department. Of the 25 children (22 male, average 10.8 years), 88% had prior history of rhinoconjunctivitis, 76% asthma, 48% eczema and 36% food allergy. After evaluation, we identified in 76% and 92% of patients food and aeroallergen sensitization, respectively; 68% had simultaneously food and inhalant sensitization and 96% had at least one positive test to aeroallergens or food allergens. The first (44%) and the most frequent (56%) symptom was dysphagia. The time between symptoms onset and the EoE diagnosis averaged 18.6 ± 29.4 months. A multidisciplinary approach is needed for a correct evaluation, intervention and follow-up of these patients.
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Esofagitis Eosinofílica/epidemiología , Adolescente , Distribución por Edad , Niño , Preescolar , Dermatitis Atópica/epidemiología , Esofagitis Eosinofílica/diagnóstico , Esofagitis Eosinofílica/inmunología , Femenino , Hipersensibilidad a los Alimentos/epidemiología , Humanos , Pruebas Inmunológicas , Lactante , Recién Nacido , Masculino , Portugal/epidemiología , Valor Predictivo de las Pruebas , Hipersensibilidad Respiratoria/epidemiología , Estudios Retrospectivos , Factores de TiempoRESUMEN
A mycelial formulation in sodium alginate pellets of the nematophagous fungus Monacrosporium thaumasium (isolate NF34A) was assessed in the biological control of beef cattle trichostrongyles in tropical Brazil. Two groups of ten male Nellore calves aged 6 months, a fungus-treated group and a control group, were fed on a pasture of Brachiaria decumbens naturally infected with larvae of cattle trichostrongyles. The fungus-treated group received doses of sodium alginate mycelial pellets orally (1 g pellets (0.2 g fungus)/10 kg live weight) twice a week for 12 months. At the end of the study there was a signiï¬cant reduction (P< 0.01) in the number of eggs per gram of faeces and coprocultures of the fungus-treated group--47.8% and 50.2%, respectively--in relation to the control group. There was a 47.3% reduction in herbage samples, collected up to 0-20 cm from faecal pats, between the fungus-treated and control groups, and a 58% reduction when the sampling distance was 20-40 cm from faecal pats (P< 0.01). The treatment with sodium alginate pellets containing the nematode-trapping fungus M. thaumasium reduced trichostrongyles in tropical south-eastern Brazil and could be an effective tool for the biological control of this parasitic nematode in beef cattle. However, in such a tropical climate with low rainfall the fungal viability can be reduced.
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Ascomicetos/fisiología , Enfermedades de los Bovinos/parasitología , Nematodos/microbiología , Infecciones por Nematodos/veterinaria , Control Biológico de Vectores/métodos , Animales , Ascomicetos/genética , Ascomicetos/aislamiento & purificación , Terapia Biológica , Brasil , Bovinos , Enfermedades de los Bovinos/terapia , Masculino , Nematodos/fisiología , Infecciones por Nematodos/parasitología , Infecciones por Nematodos/terapiaRESUMEN
The secreted polypeptide noggin (encoded by the Nog gene) binds and inactivates members of the transforming growth factor beta superfamily of signalling proteins (TGFbeta-FMs), such as BMP4 (ref. 1). By diffusing through extracellular matrices more efficiently than TGFbeta-FMs, noggin may have a principal role in creating morphogenic gradients. During mouse embryogenesis, Nog is expressed at multiple sites, including developing bones. Nog-/- mice die at birth from multiple defects that include bony fusion of the appendicular skeleton. We have identified five dominant human NOG mutations in unrelated families segregating proximal symphalangism (SYM1; OMIM 185800) and a de novo mutation in a patient with unaffected parents. We also found a dominant NOG mutation in a family segregating multiple synostoses syndrome (SYNS1; OMIM 186500); both SYM1 and SYNS1 have multiple joint fusion as their principal feature. All seven NOG mutations alter evolutionarily conserved amino acid residues. The findings reported here confirm that NOG is essential for joint formation and suggest that NOG requirements during skeletogenesis differ between species and between specific skeletal elements within species.
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Anomalías Múltiples/genética , Articulaciones/anomalías , Mutación , Proteínas/genética , Sinostosis/genética , Adolescente , Animales , Proteínas Portadoras , Gatos , Pollos , Mapeo Cromosómico , Femenino , Articulaciones de los Dedos/anomalías , Regulación del Desarrollo de la Expresión Génica , Marcadores Genéticos , Gorilla gorilla , Heterocigoto , Humanos , Articulaciones/fisiología , Masculino , Ratones , Datos de Secuencia Molecular , Morfogénesis , Análisis de Secuencia , Homología de Secuencia de Aminoácido , Homología de Secuencia de Ácido Nucleico , Porcinos , Xenopus laevis , Pez CebraRESUMEN
Altered growth and function of synoviocytes, the intimal cells which line joint cavities and tendon sheaths, occur in a number of skeletal diseases. Hyperplasia of synoviocytes is found in both rheumatoid arthritis and osteoarthritis, despite differences in the underlying aetiologies of the two disorders. We have studied the autosomal recessive disorder camptodactyly-arthropathy-coxa vara-pericarditis syndrome (CACP; MIM 208250) to identify biological pathways that lead to synoviocyte hyperplasia, the principal pathological feature of this syndrome. Using a positional-candidate approach, we identified mutations in a gene (CACP) encoding a secreted proteoglycan as the cause of CACP. The CACP protein, which has previously been identified as both 'megakaryocyte stimulating factor precursor' and 'superficial zone protein', contains domains that have homology to somatomedin B, heparin-binding proteins, mucins and haemopexins. In addition to expression in joint synovium and cartilage, CACP is expressed in non-skeletal tissues including liver and pericardium. The similarity of CACP sequence to that of other protein families and the expression of CACP in non-skeletal tissues suggest it may have diverse biological activities.
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Artropatías/genética , Pericarditis/genética , Proteoglicanos/genética , Proteoglicanos/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Bovinos , Análisis Mutacional de ADN , Femenino , Genotipo , Humanos , Hiperplasia/genética , Hiperplasia/patología , Artropatías/patología , Masculino , Datos de Secuencia Molecular , Mutación , Pericarditis/patología , Fenotipo , Proteoglicanos/química , ARN Mensajero/análisis , ARN Mensajero/genética , Homología de Secuencia de Aminoácido , Síndrome , Membrana Sinovial/metabolismo , Membrana Sinovial/patologíaRESUMEN
Coenosia attenuata, usually known as tiger-fly, is a generalist predator of agricultural and forest pests in both larval and adult stages; it has potential to be an effective biocontrol agent in protected crops. To contribute for the knowledge of which prey this predator larvae potentially consumes, and of the occurrence and the conditions that promote cannibalism by tiger-fly larvae, intact alive specimens and portions of the earthworm Lumbricus terrestris were tested as prey and the cannibalism was evaluated in the presence or in absence of fungus gnat larvae. The tiger-fly larvae fed on the bisected earthworm portions but seem to have difficulty to penetrate in the cuticle of the alive and moving L. terrestris. However, the time to start feeding on the portions of L terrestris was shorter than on fungus gnats. Cannibalism by C. attenuato was not detected, but mortality occurred in several modalities. Nevertheless, escaping from the Petri dishes was the dominant behaviour of the larvae in the cannibalism evaluation assay.
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Dípteros/fisiología , Larva/fisiología , Animales , Dípteros/crecimiento & desarrollo , Conducta Alimentaria , Larva/crecimiento & desarrollo , Oligoquetos/parasitología , Conducta PredatoriaRESUMEN
Bird migratory journeys are often long and hostile, requiring high energetic expenditure, and thus forcing birds to pause between migratory flights. Stopover sites allow migrants to replenish fuel reserves and rest, being crucial for the success of migration. Worldwide, the increasing accumulation of waste on landfills and rubbish dumps has been described to provide superabundant food resources for many bird species not only during the breeding and wintering seasons but also during migration, being used as stopover sites. Using GPS-tracking data of juvenile white storks (Ciconia ciconia) during their first migration from the Iberia Peninsula to the sub-Saharan wintering grounds, we uncover the effects of stopping en route on individual migratory performance. Particularly, we examine the benefits of stopping at artificial sites (landfills and rubbish dumps) when compared to natural stopover sites (wetlands, agricultural or desert areas) and explore the influence of anthropogenic food resources on storks' migratory strategies. Overall, white storks spent up to one-third of the migration in stopovers. We found that birds that stopped for longer periods made more detours, increasing migration duration by half a day for each stopover day. Stopping more often did not reflect on increasing in-flight energetic efficiency nor the likelihood of completing the migration. Juvenile storks used artificial sites in 80 % of the stopover days, spending 45 % less time and 10 % less energy foraging than when using natural stopovers. While stopping in landfills did not translate into differences in migratory performance, individuals in poor body condition possibly rely on these sites to improve body weight before proceeding, enabling them to successfully complete migration. Artificial stopover sites are attractive and likely increase the number and duration of stops for white storks. Even though the consequences of arriving late at the wintering grounds are unknown, it can lead to cascading consequences, influencing individual fitness and population dynamics.
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Migración Animal , Ejercicio Físico , Humanos , África del NorteRESUMEN
The Abrolhos bank is home of the richest coral reef system of the Southwestern Atlantic, where endemic coral species are found. It has been reported that Abrolhos' corals are under intense stress due to increasing of Marine Heat Waves during the last decades. Additionally, anthropic interventions along the adjacent coastal regions are a factor of concern since they contribute to the increase in the sediment load and to organic debris input in the reef domain. In November 2015, the collapse of the Fundão mining tailings dam resulted in the release of approximately 50 million m3 of iron oxide and quartz-rich slurry into the Doce River. Aiming at using a fingerprint of the tailings and to assess the presence of traces of the Fundão dam material from this event on the Abrolhos bank, this work presents new 87Sr/86Sr and 143Nd/144Nd isotope ratios of marine suspended sediment samples collected between 2016 and 2020 from a network of sediment traps throughout the reef and complementary suspended material at sea. In parallel, we monitored meteo-oceanographic parameters and modeled surface marine currents as an attempt to identify the sediment transport between the Doce River mouth and Abrolhos bank. The r isotopes were used as provenance proxies based on the fact that minerals and rocks tend to have specific isotopic signatures reflecting their own geological derivation. In this context, the isotopic ratios of various potential regional sources for the sedimentation in Abrolhos bank were evaluated. Our monitoring and isotopic measurements indicate that Doce River signatures are detected at Abrolhos bank, following the seasonal Doce River discharge at sea. Isotopic signature of Doce River at Abrolhos bank was also observed during the austral winter (July-August) when cold fronts migrate at the Brazilian coast with higher frequency and energy.
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Antozoos , Colapso de la Estructura , Animales , Brasil , Arrecifes de Coral , Sedimentos Geológicos , IsótoposRESUMEN
Climate change is predicted to severely impact interactions between prey, predators and habitats. In Southern Europe, within the Mediterranean climate, herbaceous vegetation achieves its maximum growth in middle spring followed by a three-month dry summer, limiting prey availability for insectivorous birds. Lesser kestrels (Falco naumanni) breed in a time-window that matches the nestling-rearing period with the peak abundance of grasshoppers and forecasted climate change may impact reproductive success through changes in prey availability and abundance. We used Normalised Difference Vegetation Index (NDVI) as a surrogate of habitat quality and prey availability to investigate the impacts of forecasted climate change and extreme climatic events on lesser kestrel breeding performance. First, using 14 years of data from 15 colonies in Southwestern Iberia, we linked fledging success and climatic variables with NDVI, and secondly, based on these relationships and according to climatic scenarios for 2050 and 2070, forecasted NDVI and fledging success. Finally, we evaluated how fledging success was influenced by drought events since 2004. Despite predicting a decrease in vegetation greenness in lesser kestrel foraging areas during spring, we found no impacts of predicted gradual rise in temperature and decline in precipitation on their fledging success. Notwithstanding, we found a decrease of 12% in offspring survival associated with drought events, suggesting that a higher frequency of droughts might, in the future, jeopardize the recent recovery of the European population. Here, we show that extreme events, such as droughts, can have more significant impacts on species than gradual climatic changes, especially in regions like the Mediterranean Basin, a biodiversity and climate change hotspot.
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In this work, 960 réis coins from the period when Brazil was a colony of Portugal were analyzed using the x-ray fluorescence (XRF) spectrometry. The history of these coins, dated between the end of the 17th century and the beginning of the 19th century, had a great influence on the immigration of the Portuguese Prince Regent D. João to Brazil, who arrived in 1808. Bearing in mind the need to expand the timid Brazilian monetary system, the Portuguese crown decided to collect Spanish silver pesos of 8 reales, recoined with a value of 960 réis. The recoinage procedure was carried out using a stamp; therefore, in many cases, it is possible to check the base currency. In this work, were investigated 17 samples of 960 réis coins by XRF, in which the base coin was 8 reales manufactured with raw materials from Mexican mines. In addition to characterizing the elemental composition of the coins, the XRF data were evaluated using multivariate statistical method of Robust Principal Component Analysis (ROBPCA), which was used to classify the coins based on their elemental composition. However, with XRF, elementary information is obtained for a depth of only a few micrometers. One of the essential issues in Ag-Cu metal alloys is the Ag enrichment, which can cause changes to the elemental composition of the surface. Therefore, initially, a study was carried out to verify whether the surface compositions of the coins were altered by the Ag enrichment.
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Mutations in the gene that encodes Fibroblast Growth Factor Receptor 3 (FGFR3) are associated with Achondroplasia (MIM 100800), Hypochondroplasia (MIM 146000), Muenke Syndrome (MIM 602849), Thanatophoric Dysplasia (MIM 187600, MIM 187601) and Lacrimo-Auriculo-Dento-Digital Syndrome (MIM 149730).Here we report a clinical and molecular study in a large cohort of 125 Portuguese patients with these skeletal disorders. The identification of the P250R mutation allowed the confirmation of the Muenke Syndrome in 9 out of the 52 cases referred. Two known mutations were found in the Thanatophoric Dysplasia referred cases. No mutations were identified in the LADD syndrome patient. In Achondroplasia and Hypochondroplasia, genetic heterogeneity was present amongst the 70 clinically diagnosed patients with 5 different mutations identified. As in other studies, complex phenotypic heterogeneity amongst patients carrying the same gene defect was observed. In several cases, the new amino acids encoded, as a consequence of mutations, were related to the severity of patients' phenotype. The presence of 10 misdiagnosed cases emphasizes the importance of performing mutation analysis of the hotspot regions responsible for both dysplasias (Ach and Hch). For patients with an unquestionable clinical diagnosis, lacking the most common mutations, a complete screening of FGFR3 is necessary.
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Anomalías Musculoesqueléticas/diagnóstico , Anomalías Musculoesqueléticas/genética , Mutación , Osteocondrodisplasias/diagnóstico , Osteocondrodisplasias/genética , Receptor Tipo 3 de Factor de Crecimiento de Fibroblastos/genética , Acondroplasia/diagnóstico , Acondroplasia/genética , Adolescente , Adulto , Anciano , Preescolar , Estudios de Cohortes , Femenino , Humanos , Masculino , Persona de Mediana Edad , Fenotipo , Portugal , Displasia Tanatofórica/diagnóstico , Displasia Tanatofórica/genéticaRESUMEN
The formation of scales consists in one of the most relevant problems in the oil prospecting field and occurs when incompatible types of water (injected sea water and formation water) are mixed in the reservoir, unavoidably undergoing chemical interaction followed by mineral precipitation. In this work, scale samples extracted from obstructed oil pipes were characterized through X-Ray Microfluorescence and X-Ray microtomography by analyzing their elemental and structural composition. Different types of scale were found according to their elemental distribution (mainly BaSO4 and CaCO3) and to the way that they were deposited inside the pipes. The results of both techniques provided data that can be used to optimize the prevention and removal methods of such materials from pipes and equipments used in oil facilities.
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In this study, Nerium oleander L. leaves were used as biomonitors to evaluate environmental pollutants levels in a sub-region in the Metropolitan Area of Rio de Janeiro City (Brazil) through X-ray Fluorescence (XRF). Control samples were collected in a rural zone next to the Parque Estadual da Pedra Branca/RJ. The samples were collected during all seasons of 2015 (summer, fall, winter and spring). The concentration of 13 elements (S, Cl, K, Ca, Mn, Fe, Cu, Zn, Br, Rb, Sr, Ba and Pb) was determined. Most of the elements showed, during all seasons, median concentrations of sampling sites higher than the results obtained from the control sites. The principal component analysis (PCA) showed the grouping of the elements in two main factors that can be associated to emission sources of these elements. The elements Cl, K, Ca, Cu, Rb and Sr can be associated as a possible influence from the soil (root uptake and/or resuspension). On the other hand, the elements Fe, Cu, Zn, and Pb can be associated to vehicle and industry emission sources. The study showed that the XRF technique applied to Nerium oleander L. leaves is efficient to environmental pollution analysis in Metropolitan Regions since it is precise, fast and low-cost, besides allowing the monitoring of pollution levels over time.
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Contaminación del Aire/análisis , Metales Pesados/análisis , Nerium/química , Hojas de la Planta/química , Brasil , Ciudades , Monitoreo del Ambiente/métodos , Espectrometría por Rayos XRESUMEN
Extracellular lipase from Yarrowia lipolytica was immobilized by ionotropic gelation with alginate and chitosan as encapsulating agents. Photomicrographs revealed a collapsed and heterogeneous surface of these microcapsules due to freeze-drying process. The optimum reaction temperature for the microencapsulated lipase (40⯰C) was higher than for free lipase (35⯰C) as well as the optimum pH (8.0 and 7.5, respectively). The study of the reaction kinetics showed that a higher maximum reaction rate (Vmax) (221.1â¯U/mg) for the free lipase in comparison to the immobilized form (175.3â¯U/mg). A protective effect of the microcapsule was detected in the storage of the enzyme at room temperature, as after 75â¯days 35% of activity was maintained for the microcapsules, while no activity remained after 15â¯days with the free enzyme. Lower values for inactivation constant (kd) and increase in half-life for immobilized lipase showed that lipase microencapsulation favored the thermostability of this enzyme.
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Alginatos/química , Quitosano/química , Lipasa/química , Yarrowia/enzimología , Cápsulas , Catálisis , Estabilidad de Enzimas/efectos de los fármacos , Enzimas Inmovilizadas/química , Liofilización , Concentración de Iones de Hidrógeno , Microbiología Industrial , Cinética , Microscopía Electrónica de Rastreo , Polímeros/química , Porosidad , Espectroscopía Infrarroja por Transformada de Fourier , Temperatura , Difracción de Rayos XRESUMEN
The assessment of the capacity of antivenoms to neutralize the lethal activity of snake venoms still relies on traditional rodent in vivo lethality assay. ED50 and LD50 assays require large quantities of venoms and antivenoms, and besides leading to animal suffering. Therefore, in vitro tests should be introduced for assessing antivenom neutralizing capacity in intermediary steps of antivenom production. This task is facilitated when one key lethal toxin is identified. A good example is crotoxin, a ß-neurotoxin phospholipase A2-like toxin that presents anticoagulant activity in vitro and is responsible for the lethality of venoms of Crotalus durissus snakes. By using rotational thromboelastometry, we reported recently one sensitive coagulation assay for assessing relative potency of the anti-bothropic serum in neutralizing procoagulant activity of Bothrops jararaca venom upon recalcified factor-XII-deficient chicken plasma samples (CPS). In this study, we stablished conditions for determining relative potency of four batches of the anti-crotalic serum (ACS) (antagonist) in inactivating crotoxin anticoagulant activity in CPS (target) simultaneously treated with one classical activator of coagulation (agonists). The correlation coefficient (r) between values related the ACS potency in inactivating both in vitro crotoxin anticoagulant activity and the in vivo lethality of whole venom (ED50) was 0.94 (p valueâ¯<â¯0.05). In conclusion, slowness in spontaneous thrombin/fibrin generation even after recalcification elicit time lapse sufficient for elaboration of one dose-response curve to pro- or anti-coagulant agonists in CPS. We propose this methodology as an alternative and sensitive assay for assessing antivenom neutralizing ability in plasma of immunized horses as well as for in-process quality control.
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Antivenenos/farmacología , Venenos de Crotálidos/toxicidad , Crotalus , Crotoxina/toxicidad , Tromboelastografía/métodos , Animales , Coagulación Sanguínea/efectos de los fármacos , Pollos , Venenos de Crotálidos/inmunología , Caballos , Pruebas de NeutralizaciónRESUMEN
This study used Raman, FT-IR and XRF spectroscopy and SEM to analyze ceramic fragments dating from the 19th century, excavated from an old farm in the municipality of Pirenópolis, Goiás, Brazil. The results show that the samples were produced in an open oven at a firing temperature below 500°C, using raw materials including kaolinite, hematite, magnetite, quartz, microcline, albite, anhydrite, calcite, illite, orthoclase and MnO2. Although the analyses showed similarities in the manufacturing process and the presence of many minerals was common in all samples, multivariate statistical methods (PCA) allowed a more detailed assessment of similarities and differences in the mineral composition of the samples. The results of the PCA showed that the samples excavated in one of the slave quarters (senzalas) group with those excavated at the farmhouse, where the landowner lived, which indicates a paternalistic attitude towards captives, including the sharing of ceramic materials of everyday use.
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Loxosceles gaucho spider venom induces in vitro platelet activation and marked thrombocytopenia in rabbits. Herein, we investigated the involvement of platelets in the development of the dermonecrosis induced by L. gaucho venom, using thrombocytopenic rabbits as a model. L. gaucho venom evoked a drop in platelet and neutrophil counts 4 h after venom injection. Ecchymotic areas at the site of venom inoculation were noticed as soon as 4 h in thrombocytopenic animals but not in animals with initial normal platelet counts. After 5 days, areas of scars in thrombocytopenic animals were also larger, evidencing the marked development of lesions in the condition of thrombocytopenia. Histologically, local hemorrhage, collagen fiber disorganization, and edema were more severe in thrombocytopenic animals. Leukocyte infiltration, predominantly due to polymorphonuclears, was observed in the presence or not of thrombocytopenia. Thrombus formation was demonstrated by immunohistochemistry at the microvasculature, and it occurred even under marked thrombocytopenia. Taken together, platelets have an important role in minimizing not only the hemorrhagic phenomena but also the inflammatory and wound-healing processes, suggesting that cutaneous loxoscelism may be aggravated under thrombocytopenic conditions.