Potential use of high-resolution melting analyses for SARS-CoV-2 genomic surveillance.

The pandemic caused by COVID-19 and the emergence of new variants of SARS-CoV-2 have generated clinical and epidemiological impacts on a global scale. The use of strategies for monitoring viral circulation and identifying mutations in genomic regions involved in host interaction are important measures to mitigate viral dissemination and reduce its likely complications on population health. In this context, the objective of this work was to explore the potential of high-resolution melting (HRM) analysis combined with one-step real-time reverse transcription PCR in a closed-tube system, as a fast and convenient method of screening for SARS-CoV-2 mutations with possible implications on host-pathogen interactions. The HRM analyses allowed the distinction of the Gamma, Zeta, Alpha, Delta, and Omicron variants against the predecessors (B.1.1.28, B.1.1.33) of occurrence in Brazil. It is concluded that the molecular tool standardized here has the potential to optimize the genomic surveillance of SARS-CoV-2, and could be adapted for genomic surveillance of other pathogens, due to its ability to detect, prior to sequencing, samples suggestive of new variants, selecting them more assertively and earlier for whole genome sequencing when compared to random screening.

Emergence of Dengue Virus Serotype 2 Cosmopolitan Genotype, Brazil.

We used nanopore sequencing and phylogenetic analyses to identify a cosmopolitan genotype of dengue virus serotype 2 that was isolated from a 56-year-old male patient from the state of Goiás in Brazil. The emergence of a cosmopolitan genotype in Brazil will require risk assessment and surveillance to reduce epidemic potential.

Influence of SARS-CoV-2 inactivation by different chemical reagents on the humoral response evaluated in a murine model.

Viral inactivation for antibody induction purposes, among other applications, should ensure biosafety, completely avoiding the risk of infectivity, and preserving viral immunogenicity. ß-propiolactone (BPL) is one of the most used reagents for viral inactivation, despite its high toxicity and recent difficulties related to importation, experienced in Brazil during the SARS-CoV-2 pandemic. In this context, the main objectives of this work were to test different inactivation procedures for SARS-CoV-2 and to evaluate the induction of neutralizing antibodies in mice immunized with antigenic preparations obtained after viral treatment with formaldehyde (FDE), glutaraldehyde (GDE), peroxide hydrogen (H2O2), as well as with viral proteins extract (VPE), in parallel with BPL. Verification of viral inactivation was performed by subsequent incubations of the inactivated virus in Vero cells, followed by cytopathic effect and lysis plaques observation, as well as by quantification of RNA load using reverse transcription-quantitative real time polymerase chain reaction. Once viral inactivation was confirmed, cell culture supernatants were concentrated and purified. In addition, an aliquot inactivated by BPL was also subjected to viral protein extraction (VPE). The different antigens were prepared using a previously developed microemulsion as adjuvant, and were administered in a four-dose immunization protocol. Antibody production was comparatively evaluated by ELISA and Plaque Reduction Neutralization Tests (PRNT). All immunogens evaluated showed some level of IgG anti-SARS-CoV-2 antibodies in the ELISA assay, with the highest levels presented by the group immunized with FDE-inactivated viral antigen. In the PRNT results, except for VPE-antigen, all other immunogens evaluated induced some level of neutralizing anti-SARS-CoV-2 antibodies, and the FDE-antigen stood out again with the most expressive values. Taken together, the present work shows that FDE can be an efficient and affordable alternative to BPL for the production of inactivated SARS-CoV-2 viral antigen.

Searching for plant-derived antivirals against dengue virus and Zika virus.

BACKGROUND: The worldwide epidemics of diseases as dengue and Zika have triggered an intense effort to repurpose drugs and search for novel antivirals to treat patients as no approved drugs for these diseases are currently available. Our aim was to screen plant-derived extracts to identify and isolate compounds with antiviral properties against dengue virus (DENV) and Zika virus (ZIKV). METHODS: Seven thousand plant extracts were screened in vitro for their antiviral properties against DENV-2 and ZIKV by their viral cytopathic effect reduction followed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method, previously validated for this purpose. Selected extracts were submitted to bioactivity-guided fractionation using high- and ultrahigh-pressure liquid chromatography. In parallel, high-resolution mass spectrometric data (MSn) were collected from each fraction, allowing compounds into the active fractions to be tracked in subsequent fractionation procedures. The virucidal activity of extracts and compounds was assessed by using the plaque reduction assay. EC50 and CC50 were determined by dose response experiments, and the ratio (EC50/CC50) was used as a selectivity index (SI) to measure the antiviral vs. cytotoxic activity. Purified compounds were used in nuclear magnetic resonance spectroscopy to identify their chemical structures. Two compounds were associated in different proportions and submitted to bioassays against both viruses to investigate possible synergy. In silico prediction of the pharmacokinetic and toxicity (ADMET) properties of the antiviral compounds were calculated using the pkCSM platform. RESULTS: We detected antiviral activity against DENV-2 and ZIKV in 21 extracts obtained from 15 plant species. Hippeastrum (Amaryllidaceae) was the most represented genus, affording seven active extracts. Bioactivity-guided fractionation of several extracts led to the purification of lycorine, pretazettine, narciclasine, and narciclasine-4-O-ß-D-xylopyranoside (NXP). Another 16 compounds were identified in active fractions. Association of lycorine and pretazettine did not improve their antiviral activity against DENV-2 and neither to ZIKV. ADMET prediction suggested that these four compounds may have a good metabolism and no mutagenic toxicity. Predicted oral absorption, distribution, and excretion parameters of lycorine and pretazettine indicate them as candidates to be tested in animal models. CONCLUSIONS: Our results showed that plant extracts, especially those from the Hippeastrum genus, can be a valuable source of antiviral compounds against ZIKV and DENV-2. The majority of compounds identified have never been previously described for their activity against ZIKV and other viruses.

Potential Use of DMSA-Containing Iron Oxide Nanoparticles as Magnetic Vehicles against the COVID-19 Disease.

Iron oxide magnetic nanoparticles have been employed as potential vehicles for a large number of biomedical applications, such as drug delivery. This article describes the synthesis, characterization and in vitro cytotoxic in COVID-19 cells evaluation of DMSA superparamagnetic iron oxide magnetic nanoparticles. Magnetite (Fe3O4) nanoparticles were synthesized by co-precipitation of iron salts and coated with meso-2,3-dimercaptosuccinic acid (DMSA) molecule. Structural and morphological characterizations were performed by X-ray diffraction (XRD), Fourier transformed infrared (FT-IR), magnetic measurements (SQUID), transmission electron microscopy (TEM), and dynamic light scattering (DLS). Our results demonstrate that the nanoparticles have a mean diameter of 12 nm in the solid-state and are superparamagnetic at room temperature. There is no toxicity of SPIONS-DMSA under the cells of patients with COVID-19. Taken together the results show that DMSA- Fe3O4 are good candidates as nanocarriers in the alternative treatment of studied cells.

Diversity, ecology, and bioprospecting of culturable fungi in lakes impacted by anthropogenic activities in Maritime Antarctica.

In this study, we accessed culturable fungal assemblages present in the sediments of three lakes potentially impacted anthropogenically in the Fildes Peninsula, King George Island, Antarctica and identified 63 taxa. Cladosporium sp. 2, Pseudeurotium hygrophilum, and Pseudogymnoascus verrucosus were recovered from the sampled sediments of all lakes. High concentrations of metals and the lowest fungal diversity indices were detected in the sediments of the Central Lake, which can be influenced by human activities due to their proximity to research stations to those of the other two lakes, which were far from the Antarctic stations. At least one type of biological activity was demonstrated by 40 fungal extracts. Among these, P. hygrophilum, P. verrucosus, Penicillium glabrum, and Penicillium solitum demonstrated strong trypanocidal, herbicidal, and antifungal activities. Our results suggest that an increase of the anthropogenic activities in the region might have affected the microbial diversity and composition. In addition, the fungal diversity in these lakes may be a useful model to study the effect of anthropogenic activities in Antarctica. We isolated a diverse group of fungal taxa from Antarctic lake sediments, which have the potential to produce novel compounds for the both the medical and agriculture sectors.

Diversity and bioprospecting of cultivable fungal assemblages in sediments of lakes in the Antarctic Peninsula.

We recovered 195 fungal isolates from the sediments of different lakes in the Antarctic Peninsula, which were screened to detect bioactive compounds. Forty-two taxa belonging to the phyla Ascomycota, Basidiomycota, and Mortierellomycota were identified. Thelebolus globosus, Antarctomyces psychrotrophicus, Pseudogymnoascus verrucosus, Vishniacozyma victoriae, and Phenoliferia sp. were found to be the most prevalent. The fungal assemblages showed high diversity and richness, but low dominance values. However, the diversity indices and fungal distribution ranged according to the different lake sediments. Sixty fungal extracts displayed at least one biological activity against the evaluated targets. Among them, Pseudogymnoascus destructans showed selective trypanocidal activity, Cladosporium sp. 1 and Trichoderma polysporum showed antifungal activity, and Pseudogymnoascus appendiculatus and Helotiales sp. showed high herbicidal activity. We detected a rich and diverse fungal community composed of cold cosmopolitan and psychrophilic endemic taxa recognized as decomposers, symbiotics, pathogens, and potential new species, in the sediments of Antarctic lakes. The dynamics and balance of this fungal community represents an interesting aquatic web model for further ecological and evolutionary studies under extreme conditions and potential climate changes in the regions. In addition, we detected fungal taxa and isolates able to produce bioactive compounds that may represent the source of prototype molecules for applications in medicine and agriculture.

Cultivable fungi present in deep-sea sediments of Antarctica: taxonomy, diversity, and bioprospecting of bioactive compounds.

We accessed the culturable mycobiota present in marine sediments at different depths in Antarctica Ocean. Acremonium fusidioides, Penicillium allii-sativi, Penicillium chrysogenum, Penicillium palitans, Penicillium solitum, and Pseudogymnoascus verrucosus were identified. Penicillium allii-sativi was the dominant species. At least one isolate of each species was capable to present antifungal, trypanocidal, leishmanicidal, antimalarial, nematocidal, or herbicidal activities. Penicillium produced extracts with strong trypanocidal and antimalarial activities, and the extracts of P. solitum and P. chrysogenum demonstrated strong antimalarial activities. Acremonium fusidioides and P. verrucosus displayed strong selective herbicidal properties. The 1H NMR signals for extracts of A. fusidioides, P. chrysogenum, and P. solitum indicated the presence of highly functionalized secondary metabolites, which may be responsible for the biological activities detected. In the deep marine Antarctic sediments, we detected fungal assemblages in which the Penicillium species were found to be dominant and demonstrated capabilities to survive and/or colonise that poly-extreme habitat. Penicillium being a polyextremophile Antarctic species, exhibited strong biological activities and the presence of aromatic compounds in its extracts may indicate that they are wild ancient strains with high genetic and biochemical potentials that enable them to produce bioactive compounds which can be researched in further studies and used in the chemotherapy of neglected tropical diseases as well as in agriculture.

Cultivable fungi present in Antarctic soils: taxonomy, phylogeny, diversity, and bioprospecting of antiparasitic and herbicidal metabolites.

Molecular biology techniques were used to identify 218 fungi from soil samples collected from four islands of Antarctica. These consisted of 22 taxa of 15 different genera belonging to the Zygomycota, Ascomycota, and Basidiomycota. Mortierella, Antarctomyces, Pseudogymnoascus, and Penicillium were the most frequently isolated genera and Penicillium tardochrysogenum, Penicillium verrucosus, Goffeauzyma gilvescens, and Mortierella sp. 2 the most abundant taxa. All fungal isolates were cultivated using solid-state fermentation to obtain their crude extracts. Pseudogymnoascus destructans, Mortierella parvispora, and Penicillium chrysogenum displayed antiparasitic activities, whilst extracts of P. destructans, Mortierella amoeboidea, Mortierella sp. 3, and P. tardochrysogenum showed herbicidal activities. Reported as pathogenic for bats, different isolates of P. destructans exhibited trypanocidal activities and herbicidal activity, and may be a source of bioactive molecules to be considered for chemotherapy against neglected tropical diseases. The abundant presence of P. destructans in soils of the four islands gives evidence supporting that soils in the Antarctic Peninsula constitute a natural source of strains of this genus, including some P. destructans strains that are phylogenetically close to those that infect bats in North America and Europe/Palearctic Asia.

Fungi associated with rocks of the Atacama Desert: taxonomy, distribution, diversity, ecology and bioprospection for bioactive compounds.

This study assessed the diversity of cultivable rock-associated fungi from Atacama Desert. A total of 81 fungal isolates obtained were identified as 29 Ascomycota taxa by sequencing different regions of DNA. Cladosporium halotolerans, Penicillium chrysogenum and Penicillium cf. citrinum were the most frequent species, which occur at least in four different altitudes. The diversity and similarity indices ranged in the fungal communities across the latitudinal gradient. The Fisher-α index displayed the higher values for the fungal communities obtained from the siltstone and fine matrix of pyroclastic rocks with finer grain size, which are more degraded. A total of 23 fungal extracts displayed activity against the different targets screened. The extract of P. chrysogenum afforded the compounds α-linolenic acid and ergosterol endoperoxide, which were active against Cryptococcus neoformans and methicillin-resistance Staphylococcus aureus respectively. Our study represents the first report of a new habitat of fungi associated with rocks of the Atacama Desert and indicated the presence of interesting fungal community, including species related with saprobes, parasite/pathogen and mycotoxigenic taxa. The geological characteristics of the rocks, associated with the presence of rich resident/resilient fungal communities suggests that the rocks may provide a favourable microenvironment fungal colonization, survival and dispersal in extreme conditions.

Diversity and bioprospection of fungal community present in oligotrophic soil of continental Antarctica.

We surveyed the diversity and capability of producing bioactive compounds from a cultivable fungal community isolated from oligotrophic soil of continental Antarctica. A total of 115 fungal isolates were obtained and identified in 11 taxa of Aspergillus, Debaryomyces, Cladosporium, Pseudogymnoascus, Penicillium and Hypocreales. The fungal community showed low diversity and richness, and high dominance indices. The extracts of Aspergillus sydowii, Penicillium allii-sativi, Penicillium brevicompactum, Penicillium chrysogenum and Penicillium rubens possess antiviral, antibacterial, antifungal, antitumoral, herbicidal and antiprotozoal activities. Bioactive extracts were examined using (1)H NMR spectroscopy and detected the presence of secondary metabolites with chemical shifts. Our results show that the fungi present in cold-oligotrophic soil from Antarctica included few dominant species, which may have important implications for understanding eukaryotic survival in cold-arid oligotrophic soils. We hypothesize that detailed further investigations may provide a greater understanding of the evolution of Antarctic fungi and their relationships with other organisms described in that region. Additionally, different wild pristine bioactive fungal isolates found in continental Antarctic soil may represent a unique source to discover prototype molecules for use in drug and biopesticide discovery studies.

Avaliação da Atividade Antiviral de Extratos Vegetais e de Fungos contra Dengue Virus

Produtos naturais são potenciais fontes alternativas para o desenvolvimento de antivirais para o tratamento da dengue, assim como de outras doenças causadas por vírus da família Flaviviridae ou mesmo para um amplo espectro de viroses. Neste estudo foi feita a triagem da atividade in vitro contra o Dengue virus 2 (DENV-2) de 3101 extratos, provenientes de plantas e de fungos da Coleção de Amostras para Bioensaios da Fiocruz. Para tal, células BHK-21 foram infectadas com DENV-2 e tratadas simultaneamente com 25 μg/mL de extrato sendo o resultado analisado por dois métodos: observação do grau de inibição do efeito citopático (ECP) por microscopia óptica e análise da viabilidade celular pelo ensaio colorimétrico do MTT. Dentre os 3101 extratos testados, 115 extratos apresentaram atividade antiviral contra DENV-2 e foram selecionados para a determinação da respectiva concentração efetiva 50 (CE50).

Cinquenta e cinco destes extratos foram obtidos de plantas pertencentes a 20 famílias distintas: Amaryllidaceae (3), Annonaceae (1), Asteraceae (5), Begoniaceae (1), Clusiaceae (1), Combretaceae (1), Erythroxylaceae (1), Fabaceae (4), Lythraceae (2), Malpighiaceae (8), Malvaceae(1), Melastomataceae (2), Melochia (1), Myrtaceae (3), Rubiaceae (8), Sapindaceae(9), Ochnaceae (1), Primulaceae (1) Vitaceae (1), Vochysiaceae (1). Os demais extratos (60) foram obtidos de culturas de fungos endofíticos coletados no Brasil, no continente Antártico e no Deserto do Atacama, ainda não identificados. Até o momento, os extratos vegetais mais promissores foram obtidos de plantas da família Amaryllidaceae (IS = 32,15) e da família Fabaceae (IS = 20,47) e (IS = 24,47). Vinte extratos fúngicos apresentaram valores de CE50 que variaram entre 3,1 a 12,5 μg/mL e sem citotoxicidade aparente até a concentração de 100 μg/mL. Nossos resultados mostram que tais plantas e fungos são fontes promissoras de substâncias com ação antiviral contra DENV.

Supratentorial primary neuroectodermal tumor (PNET) inside the third ventricle in adult: an rare case report / Tumor neuroectodérmico primário (PNET) supratentorial no terceiro ventrículo em adulto: um raro relato de caso

The PNET of CNS are considered malignant undifferentiated tumors, and it represents about 2,8% of all tumors found on infants and teenagers, more rarely found on adults. In the present article will report the case of a patient, male, 23 years-old, with nodular lesion inside the third ventricle, admitted on emergency room with acute intense headache, drowsiness, vomiting and visual clouding, started three days before. Although there have been advances in diagnosis and treatment of PNET in children, few publications were found on the efficiency of available treatment options on adults. In our patient the lesion was completely removed by a anterosuperior interhemispheric transcallosal craniotomy, and subsequently diagnosed as PNET by anatomopathological. Postoperative hydrocephaly was installed and reverted with a ventriculoperitoneal shunt, with clinical and neurological status improvement. The patient died 18 months after diagnosis, due to respiratory hospitalar infection...

Os PNET do SNC são considerados tumores malignos indiferenciados e representam cerca de 2,8% de todos os tumores encontrados em crianças e adolescentes, sendo mais raramente encontrados em adultos. Neste presente artigo, relatamos o caso de um paciente, do sexo masculino, com 23 anos de idade, com lesão nodular dentro do terceiro ventrículo, que foi admitido na emergência com cefaleia intensa e aguda, sonolência, vômitos e turvação visual, iniciados três dias antes da admissão. Embora tenha havido avanços no diagnóstico e tratamento de PNET em crianças, poucas publicações foram encontradas sobre a eficiência de opções de tratamento disponíveis em adultos. No nosso paciente a lesão foi completamente removida por uma craniotomia ântero-transcalosa inter-hemisférica; posteriormente, ele foi diagnosticado como PNET por meio do exame anatomopatológico. A hidrocefalia instalada no pós-operatório foi revertida com uma derivação ventrículo-peritoneal, com melhora do quadro clínico e neurológico. O paciente morreu 18 meses após o diagnóstico, em decorrência de uma infecção hospitalar respiratória...

Análise pós-operatória tardia da artrodese por via posterior em pacientes ASIA/Frankel e com trauma raquimedular toracolombar em serviço universitário / Postoperative late analysis of posterior arthrodesis in patients ASIA/Frankel e with thoracolumbar spine trauma in university hospital

Objetivo: O propósito deste estudo foi avaliar retrospectivamente o tratamento cirúrgico de pacientes vítimas de trauma raquimedular desse segmento sem déficit neurológico, quanto à evolução da dor local e à deformidade cifótica local e regional do segmento acometido, em um serviço universitário. Método: Foram utilizados dados de prontuário e avaliações clínicas e radiológicas de 16 pacientes submetidos à artrodese pedicular por via posterior, no período de maio de 2003 a dezembro de 2006, operados há pelo menos dois anos. Resultados: A média de idade foi de 43,43 ± 11,44 anos e o nível mais acometido L1. O mecanismo principal do trauma foi queda de altura. Após realização de raios X em perfil e cálculo dos ângulos local sagital (ALS) e regional sagital (ARS), de pós-operatório precoce e tardio, não houve diferença estatisticamente significativa entre os dados obtidos, apesar da diminuição de -15,3º para -10,7º nas cifoses locais e do aumento de -13,7º para 15,9º nas cifoses regionais. Houve diferença estatística na avaliação de dor pela Escala Analógica de Dor (EAD), entre o pré-operatório e o pós-operatório tardio (maior que dois anos), com redução de quatro vezes da intensidade da dor entre os pacientes analisados (p < 0,05). Conclusão: A artrodese pela via posterior é uma proposta alternativa de tratamento quando se discute a dor desses pacientes. Fica a necessidade de abranger um número maior de pacientes com essa patologia, a fim de dispor dados mais fidedignos, respaldando o tratamento como uma alternativa viável no manejo de pacientes com fraturas toracolombares e neurologicamente intactos.

Objective: The aim of this study was a retrospective valuation of surgical treatment of patients with spinal thoracolumbar spine without neurological deficit, for that local pain, local and regional kyphotic deformity at an university hospital. Method: There were used hospital datum from 16 operated patients and clinical and radiological evaluation submitted a pedicular screw instrumentation from May 2003 until December 2006, operated for at least 2 years. Results: The mean age was 43.43 ± 11.44 years and the most fractured level L1. The principal mechanism was height fall. After the x-rays realized and calculated the local sagittal angle and regional sagittal angle, there's no significative difference between them, despite the reduction of -15.3º to -10.7º, at local kyphosis and augmentation of -13.7º to 15.9º at regional kyphosis. There was statistics difference at pain evaluation using the Analogic Pain Scale, between the pre-operative and postoperative (beyond two years), with four times reduction of intensity pain among the analyzed patients (p < 0.05). Conclusion: The arthrodesis by posterior approach is an alternative proposal of treatment when discussing the pain of these patients. It is the need to include a larger number of patients with this pathology, in order to dispose more data reliable, endorsing the treatment as a viable alternative in the management of patients with thoracolumbar fractures and neurologically intact.

Fatores de risco para sibilância no primeiro ano de vida / Risk factors for wheezing in the first year of life

OBJETIVO: Verificar os fatores de risco para sibilância em lactentes no sul do Brasil. MÉTODOS: Estudo transversal, onde foram aplicados questionários padronizados e validados (Estudio Internacional de Sibilancias en Lactantes - EISL), aos pais de lactentes com idade entre 12 e 15 meses que procuraram 35 das 107 unidades de saúde para imunização rotineira no período entre agosto de 2005 e dezembro de 2006. Foi realizada análise univariada entre sibilância e os fatores estudados utilizando razão de prevalência (RP) e intervalo de confiança de 95 por cento (IC95 por cento). Fatores associados à sibilância na análise bivariada foram avaliados com a utilização de regressão de Poisson. RESULTADOS: Três mil e três lactentes participaram do estudo. Os fatores de risco para sibilância foram: gênero masculino (RP = 1,14; IC95 por cento 1,05-1,24), história familiar de asma [mãe (RP = 1,18; IC95 por cento 1,04-1,33); pai (RP = 1,20; IC95 por cento 1,05-1,39); irmãos (RP = 1,23; IC95 por cento 1,08-1,42)], outros animais domésticos (pássaros, coelhos, etc.) presentes durante a gravidez (RP = 1,28; IC95 por cento 1,07-1,53), idade de início na creche [0-3 meses (RP = 1,15; IC95 por cento 0,98-1,34); 4-6 meses (RP = 1,39; IC95 por cento 1,24-1,55); 7-12 meses (RP = 1,20; IC95 por cento 1,07-1,35)], seis ou mais episódios de resfriado (RP = 1,32; IC95 por cento 1,21-1,44), história pessoal de dermatite (RP = 1,09; IC95 por cento 1,003-1,19) e mofo no domicílio (RP = 1,14; IC95 por cento 1,04-1,24). Imunização atualizada (RP = 0,79; IC95 por cento 0,63-0,98) e banheiro no domicílio (RP = 0,83; IC95 por cento 0,68-1,01) foram fatores de proteção. CONCLUSÃO: Os fatores de risco independentes para sibilância no primeiro ano de vida também são conhecidos como risco para asma em crianças e adolescentes. Esses dados são úteis para prever o diagnóstico de asma e instituição de medidas de prevenção quando cabíveis.

OBJECTIVE: To assess risk factors for wheezing in infants in southern Brazil. METHODS: Cross-sectional study using a standardized and validated questionnaire (Estudio Internacional de Sibilancias en Lactantes - EISL, or International Study of Wheezing in Infants). Parents of infants aged 12-15 months who attended 35 of 107 health centers between August 2005 and December 2006 for regular immunization were interviewed. The association between wheezing and factors studied was made using a prevalence ratio (PR) and confidence interval of 95 percent (95 percentCI) to perform a univariate analysis. Factors associated with wheezing in the bivariate analysis were studied using Poisson regression. RESULTS: Three thousand and three parents of infants filled out the questionnaire. The risk factors were male gender (PR = 1.14; 95 percentCI 1.05-1.24), history of asthma in the family [mother (PR = 1.18; 95 percentCI 1.04-1.33); father (PR = 1.20; 95 percentCI 1.05-1.39); siblings (PR = 1.23; 95 percentCI 1.08-1.42)], other pets in the home during pregnancy (PR = 1.28; 95 percentCI 1.07-1.53), age when child started daycare [0-3 months (PR = 1.15; 95 percentCI 0.98-1.34); 4-6 months (PR = 1.39; 95 percentCI 1.24-1.55); 7-12 months (PR = 1.20; 95 percentCI 1.07-1.35)], six or more episodes of cold (PR = 1.32; 95 percentCI 1.21-1.44), personal history of dermatitis (PR = 1.09; 95 percentCI 1.003-1.19), and mold in the home (PR = 1.14; 95 percentCI 1.04-1.24). Up-to-date immunization (PR = 0.79; 95 percentCI 0.63-0.98) and bathroom in the home (PR = 0.83; 95 percentCI 0.68-1.01) were protective factors. CONCLUSIONS: Independent risk factors for wheezing in the first year of life are also known risks for asthma in children and adolescents. These data are useful to predict the diagnosis of asthma and to promote its prevention (when applicable).