Genomic diversity of the human pathogen Paracoccidioides across the South American continent.

Paracoccidioidomycosis (PCM) is a life-threatening systemic mycosis widely reported in the Gran Chaco ecosystem. The disease is caused by different species from the genus Paracoccidioides, which are all endemic to South and Central America. Here, we sequenced and analyzed 31 isolates of Paracoccidioides across South America, with particular focus on isolates from Argentina and Paraguay. The de novo sequenced isolates were compared with publicly available genomes. Phylogenetics and population genomics revealed that PCM in Argentina and Paraguay is caused by three distinct Paracoccidioides genotypes, P. brasiliensis (S1a and S1b) and P. restrepiensis (PS3). P. brasiliensis S1a isolates from Argentina are frequently associated with chronic forms of the disease. Our results suggest the existence of extensive molecular polymorphism among Paracoccidioides species, and provide a framework to begin to dissect the connection between genotypic differences in the pathogen and the clinical outcomes of the disease.

Geographical distribution and ecological niche modeling of the etiological agents of human sporotrichosis in Venezuela.

The geographical distribution and ecological niche of the two circulating species of the Sporothrix genus in Venezuela was established. For this, 68 isolates of Sporothrix spp. from patients of different regions of the country were analyzed. A molecular taxonomy analysis was conducted using a fragment of the calmodulin gene (CAL), and ITS regions, confirming the presence of S. schenckii (62%) and S. globosa (38%). Computational models of ecological niche for each species were obtained by the maximum entropy method using the MaxEnt software, which predicted the best environmental conditions for the presence of the two species. These models predict that the main variables influencing the presence of S. schenckii were altitude and annual mean temperature, while for S. globosa, the more influent variable was the land use, with 82% of S. globosa located at urban areas vs 56% for S. schenckii. The results here presented could contribute to understand the specific environmental factors that might modulate the occurrence of Sporothrix spp. as well as its transmission. To our knowledge, our analyses show for the first time Sporothrix spp.-specific ecological niche data, a valuable tool to promote evidence-based public health policymaking within endemic areas of sporotrichosis.

Comparison of Cell Wall Polysaccharide Composition and Structure Between Strains of Sporothrix schenckii and Sporothrix brasiliensis.

Sporothrix schenckii, Sporothrix brasiliensis, and Sporothrix globosa are the main causative agents of sporotrichosis, a human subcutaneous mycosis. Differences in virulence patterns are associated with each species but remain largely uncharacterized. The S. schenckii and S. brasiliensis cell wall composition and virulence are influenced by the culturing media, with little or no influence on S. globosa. By keeping constant the culturing media, we compared the cell wall composition of three S. schenckii and two S. brasiliensis strains, previously described as presenting different virulence levels on a murine model of infection. The cell wall composition of the five Sporothrix spp. strains correlated with the biochemical composition of the cell wall previously reported for the species. However, the rhamnose-to-ß-glucan ratio exhibits differences among strains, with an increase in cell wall rhamnose-to-ß-glucan ratio as their virulence increased. This relationship can be expressed mathematically, which could be an important tool for the determination of virulence in Sporothrix spp. Also, structural differences in rhamnomannan were found, with longer side chains present in strains with lower virulence reported for both species here studied, adding insight to the importance of this polysaccharide in the pathogenic process of these fungi.

Expression of Paracoccidioides brasiliensis CHS3 in a Saccharomyces cerevisiae chs3 null mutant demonstrates its functionality as a chitin synthase gene.

We report the isolation and sequencing of CHS3, a gene that encodes one of several chitin synthases in Paracoccidioides brasiliensis, a medically important fungus restricted geographically to Latin America. The gene contains a single open reading frame of 3817 bp with two introns (71 and 86 bp) and encodes a 1220 amino acid polypeptide with high similarity to other fungal chitin synthases. Northern analysis reveals a high expression of CHS3 in the pathogenic yeast-like phase of the fungus and at the end of the mycelium-yeast transition. Expression of P. brasiliensis CHS3 in a Saccharomyces cerevisiae chs3 null mutant enhanced calcofluor white staining in parallel to an increase in total chitin synthase activity and chitin content in its cell wall.

Cell wall glucan synthases and GTPases in Paracoccidioides brasiliensis.

In this report we identified orthologues of fungal AGS1, RHO1, RHO2, RAC1 and CDC42 genes in the dimorphic fungus Paracoccidioides brasiliensis. Based on its homology to known fungal sequences, P. brasiliensis Ags1 was identified as an alpha-1,3-glucan synthase, while Rho1, Rho2, Rac1 and Cdc42 proteins were classified into the Rho1, Rho2, Rac1 and Cdc42 subgroups of fungal Rho GTPases, respectively. Of them, Rho1 is one of two subunits of a putative beta-1,3-glucan synthase complex, the other being the synthase itself (Fks1), while Rho2 has been associated to the alpha-1,3-glucan synthase (Ags1). Expression studies showed that mRNAs levels of RHO2 and AGS1 kept a direct relationship but the levels of RHO1 and FKS1 did not. P. brasiliensis RHO1 successfully restored growth of Saccharomyces cerevisiae rho1 mutant under restrictive temperature conditions. Chemical analyses of P. brasiliensis alpha-1,3-glucan, synthesized by Ags1p, indicated that it is essentially a linear polysaccharide, with <3% of alpha-1,4-linked glucose branches, occasionally attached as single units to the alpha-1,3-backbone.

Cell walls of the dimorphic fungal pathogens Sporothrix schenckii and Sporothrix brasiliensis exhibit bilaminate structures and sloughing of extensive and intact layers.

Sporotrichosis is a subcutaneous mycosis caused by pathogenic species of the Sporothrix genus. A new emerging species, Sporothrix brasiliensis, is related to cat-transmitted sporotrichosis and has severe clinical manifestations. The cell wall of pathogenic fungi is a unique structure and impacts directly on the host immune response. We reveal and compare the cell wall structures of Sporothrix schenckii and S. brasiliensis using high-pressure freezing electron microscopy to study the cell wall organization of both species. To analyze the components of the cell wall, we also used infrared and 13C and 1H NMR spectroscopy and the sugar composition was determined by quantitative high-performance anion-exchange chromatography. Our ultrastructural data revealed a bi-layered cell wall structure for both species, including an external microfibrillar layer and an inner electron-dense layer. The inner and outer layers of the S. brasiliensis cell wall were thicker than those of S. schenckii, correlating with an increase in the chitin and rhamnose contents. Moreover, the outer microfibrillar layer of the S. brasiliensis cell wall had longer microfibrils interconnecting yeast cells. Distinct from those of other dimorphic fungi, the cell wall of Sporothrix spp. lacked α-glucan component. Interestingly, glycogen α-particles were identified in the cytoplasm close to the cell wall and the plasma membrane. The cell wall structure as well as the presence of glycogen α-particles varied over time during cell culture. The structural differences observed in the cell wall of these Sporothrix species seemed to impact its uptake by monocyte-derived human macrophages. The data presented here show a unique cell wall structure of S. brasiliensis and S. schenckii during the yeast parasitic phase. A new cell wall model for Sporothrix spp. is therefore proposed that suggests that these fungi molt sheets of intact cell wall layers. This observation may have significant effects on localized and disseminated immunopathology.

Dando sentido ao ensino do Brinquedo Terapêutico: a vivência de estudantes de enfermagem / Giving meaning to the teaching of Therapeutic Play: the experience of nursing students

Objetivo: Compreender o significado atribuído pelo graduando de enfermagem quanto ao ensino e à prática do Brinquedo Terapêutico (BT) no Curso de Graduação em Enfermagem. Métodos: Pesquisa qualitativa, realizada à luz do Interacionismo Simbólico. Participaram 17 estudantes de enfermagem de uma universidade pública, sendo os dados coletados por meio de entrevista semiestruturada e analisados pela Análise Qualitativa de Conteúdo Convencional. Resultados: Possibilitaram compreender o significado atribuído pelo graduando de enfermagem ao ensino do BT no curso de graduação, como uma intervenção necessária e importante para promover um cuidado de enfermagem qualificado e humano; em vista disso, seu ensino deve ser garantido aos alunos de graduação. Considerações Finais: Espera-se que seus resultados contribuam para uma reflexão sobre o ensino do BT, visando que esse conteúdo ultrapasse as fronteiras da sala de aula e da prática acadêmica, favorecendo a sensibilização do aluno para sua utilização futura, quando enfermeiro.

Ataque isquémico transitorio: incidencia de accidente cerebrovascular fatal / Transient ischemic attack: incidence of fatal stroke

El AIT (ataque isquémico transitorio) se considera predictor de ACV. El seguimiento a seis meses mostró un riesgo cercano al 10%. Son dos espectros de la enfermedad cerebro vascular isquémica de alto impacto en la morbi-mortalidad a nivel mundial. Objetivo: evaluar la incidencia de ACV en pacientes con AIT hospitalizados en dos instituciones con seguimiento a 180 días. Metodología: estudio de cohorte, el AIT se definió por historia y examen clínico, excluyendo el infarto por neuroimagen. Se evaluó el riesgo de ACV con la escala ABCD2, documentándolo por neuroimagen. Resultados: ingresaron 85 casos (abril 2012/ abril 2013). Edad promedio 68.1 años (DE 13.5), 62.4% mujeres. Los factores de riesgo más frecuentes fueron HTA (69.4%), dislipidemia (56.4%), tabaquismo (31.1%) y antecedente de ACV (18.8%). El 24.6% presentó ACV (n:19), con dos fallecimientos (2.4%). El ABCD2 ≥ 5 fue predictor de ACV, HR 4.7 [IC 95% 1.1-20.7]. Entre los pacientes con previo antecedente de ACV, la mitad de ellos repitió el evento (8/16), HR 2.2 [IC 95% 0.81-6.1]. Conclusión: hay alta incidencia de ACV después de AIT. Se requieren estudios con muestra de mayor tamaño.

Transient ischemic attacks (TIAs) are considered predictors of CVA. A six-month follow-up showed a risk of nearly 10%. There are two spectrums of ischemic cerebrovascular disorders with high impact on morbidity and mortality worldwide. Objective: to assess the incidence of CVAs after a 180-day follow-up in patients with prior TIA hospitalized in two institutions. Methodology: a cohort study: TIA was defined by medical record and physical exam, excluding infarct by neuroimaging. CVA risk was assessed using the ABCD2 scale and documented by neuroimaging. Results: eighty-five cases were admitted between April, 2012 and April, 2013; mean age was 68.1 years (SD 13.5); 62.4% were females. The most frequent risk factors were, ATH (69.4%), dyslipidemia (56.4%), smoking (31.1%) and prior stroke (18.8%). A CVA occurred in 24.6% (n: 19), accounting for 2 deaths (2.4%). An ABCD2 score ≥ 5 was a predictor of CVA, HR 4.7 [CI 95% 1.1-20.7]. Of patients with a prior CVA half presented a new event (8/16), HR 2.2 [CI 95% 0.81-6.1]. Conclusion: there is a high incidence of CVA after a TIA. Studies with a larger sample size are required.

Síncope convulsivo como primera manifestación clínica de paraganglioma carotídeo / Convulsive syncope as first clinical manifestation of carotid paraganglioma

El síncope convulsivo se define como episodio de pérdida de la conciencia con postura tónica. Hace parte de los diagnósticos diferenciales de la epilepsia, como evento paroxístico con manifestaciones motoras, clónica y/o mioclónica, sin descargas epileptiformes en el electroencefalograma. La crisis del síncope hace parte de la respuesta del cerebro frente a estados de hipoxia y representa un reto que ha generado controversia acerca del comportamiento y conducta a seguir. Se reporta el caso de un hombre de 53 años que ingresa a urgencias por presentar un único episodio sincopal, asociado con postura tónica en extensión de extremidades superiores, supraversión de la mirada y relajación de esfínteres. Los estudios de extensión mostraron una masa dependiente de la carótida izquierda en su bifurcación compatible con paraganglioma, que a la palpación generaba la aparición de eventos similares al del ingreso simulando semiología ictal...

Convulsive syncope is defined as an episode of loss of consciousness and tonic posture. It is part of epilepsy´s differential diagnoses, as a paroxysmal event with motor clonic and/or myoclonic activity, showing no epileptiform discharges in the electroencephalogram. Syncope episodes are a component of the brain´s response to hypoxia and represent a challenge which remains controversial on adequate handling and therapy. We report on a 53-year old male patient who presented to the emergency room for having experienced one syncope episode associated with tonic postures involving extension of the upper extremities, upward gaze and urine and fecal incontinence. The diagnostic work up revealed a mass in the left common carotid artery bifurcation compatible with a paraganglioma. Similar events to that of admission were triggered by palpation of the mass simulating ictal semiology...

Perfil epidemiológico dos pacientes diagnosticados com hanseníase através de exame de contato no município de Campos dos Goytacazes, RJ / Epidemiological profile of patients diagnosed with leprosy by contact exam in the municipality of Campos dos Goytacazes, RJ

JUSTIFICATIVA E OBJETIVOS: Realizou-se esse estudo para obter informações pontuais visando estabelecer e valorizar medidas de controle da cadeia epidemiológica da doença. O objetivo deste estudo foi investigar a prevalência de pacientes diagnosticados com hanseníase por exame de contato assim como o seu perfil epidemiológico. MÉTODO: Trata-se de um estudo transversal composto por 82 pacientes diagnosticados por exame de contato entre 2006 e 2010, a partir da análise das fichas de notificação do SINAN (Sistema de Informação de Agravos de Notificação) disponíveis no Programa Municipal de Controle da Hanseníase. Foi determinada a prevalência deste modo de detecção em relação ao número total de casos e as variáveis analisadas foram: sexo, idade, formas clínicas e classificação operacional. RESULTADOS: Constatou-se prevalência de 16% de pacientes detectados por exame de contato do total de 564 casos notificados. Observou-se que 51% eram do sexo feminino. A faixa etária mais acometida englobou crianças até 15 anos, correspondendo a 25% (21 casos). A classificação operacional paucibacilar foi a mais prevalente com 75% do total, sendo a forma tuberculoide presente em 50% dos casos. CONCLUSÃO: Observou-se a importância do exame de contato para interromper a cadeia epidemiológica da doença, diagnosticando precocemente sinais e sintomas e instituindo o tratamento adequado, para que possam evitar as formas graves da doença, incluindo suas incapacidades.

BACKGROUND AND OBJECTIVES: We conducted this study to obtain specific information to establish and enhance measures to control disease epidemiological chair. The objective of this study were investigated the prevalence of patients diagnosed with leprosy by examination of contact as well as the epidemiological profile of them. METHOD: This is a cross-sectional study comprising 82 patients diagnosed by examination of contact between 2006 and 2010, from the analysis of reporting forms SINAN (Information System for Notifiable Diseases) available in the Municipal Program for the Control of Leprosy. We determined the prevalence of this mode of detection in the total number of cases and the variables analyzed were gender, age, clinical forms and clinical classification. RESULTS: We found prevalence of 16% of patients detected by contact exam of the total 564 cases reported. It was observed that 51% were female. The most affected age group comprised children less than 15 years, corresponding to 25% (21 cases). The operational classification paucibacillary was most prevalent with 75% of the total, with the tuberculoid form present in 50% of cases. CONCLUSION: It may be noted the importance of this contact to stop the epidemiology of the disease, diagnosing early signsand symptoms and instituting proper treatment so they can prevent serious disease, including their disabilities the total number of cases and the variables analyzed were gender, age, clinical forms and clinical classification.

Primers for clinical detection of Paracoccidioides brasiliensis.

From a 0.72-kb fragment universally generated in Paracoccidioides brasiliensis strains, primers were designed and tested on genomic DNA of this and other pathogenic fungi. They were specific and highly sensitive for P. brasiliensis DNA. Positive results were obtained when these were tested in clinical samples.

Doença do armazenamento lisossomal induzida pelo consumo de Sida carpinifolia em bovinos do Rio Grande do Sul / Lysosomal storage disease caused by Sida carpinifolia in cattle in Rio Grande do Sul

This paper reports the natural poisoning by Sida carpinifolia (guanxuma, chá-da-índia) in cattle in Rio Grande do Sul, Brazil. Five cattle were affected in the period 2001-2008. Clinical signs included weight loss, incoordination, walking difficulty, generalized tremors, frequent falls, and death. Microscopically, the main changes were vacuolation of Purkinje neurons in the cerebellum, pancreatic acinar cells, and thyroid follicular cells. Transmission electron microscopy revealed vacuoles bordered by membrane containing finely granular material. Lectin histochemistry showed positive staining in neurons with the lectins Concanavalia ensiformis (Con-A), Triticum vulgaris (WGA), and Succinyl Triticum vulgaris (sWGA).

Relata-se a intoxicação natural por Sida carpinifolia (guanxuma, chá-da-índia) em bovinos no Rio Grande do Sul. Foram afetados cinco bovinos no período 2001-2008. O quadro clínico foi caracterizado por emagrecimento, incoordenação, dificuldade de locomoção, tremores generalizados, quedas frequentes e morte. Microscopicamente, as principais alterações foram vacuolização dos neurônios de Purkinje do cerebelo, das células acinares do pâncreas e das células foliculares da tireoide. A microscopia eletrônica evidenciou vacúolos com conteúdo finamente granulado e delimitado por membrana. Na lectina-histoquímica, observou-se marcação em neurônios com as lectinas Concanavalia ensiformis (Con-A), Triticum vulgaris (WGA) e Succinyl Triticum vulgaris (sWGA).

Anemia Hemolítica auto-imune em crianças / Autoimmune hemolytic Anemia in children

Anemia hemolítica auto-imune é uma doença rara, na qual ocorre destruição prematura das hemácias devido à presença de auto-anticorpos. Em crianças, o tipo mais comum dessa doença é a anemia hemolítica auto-imune por anticorpos da classe IgG, sendo comumente associado a outras patologias. A primeira linha de tratamento é a corticoterapia, no entanto nem todos os casos apresentam uma resposta efetiva. Na grande maioria, o prognóstico é bom, mas existem casos graves e até mesmo fatais.

Variação da capacidade residual funcional e da capacidade inspiratória após o uso do broncodilatador / Variation of functional residual capacity and inspiratory capacity with bronchodilator

Introdução: a resposta aos broncodilatadores (BD) aferida do volume expiratório forçado no primeiro segundo (VEF1) já está universalmente estabelecida, porém as variações dos volumes pulmonares como critérios de responsividade aos BD precisam ser melhor avaliadas. Métodos: os pacientes foram divididos em dois grupos: Distúrbio Ventilatório Obstrutivo (DVO) e Normais (N), pela espirometria e classificados segundo o Consenso Brasileiro de Espirometria de 1996. Posteriormente foram subdivididos em responsivos ao BD (RBD) e não responsivos (NRBD). Todos realizaram medidas dos volumes pulmonares: Volume Residual (VR), Capacidade Residual Funcional (CRF), Capacidade Inspiratória (CI) através da pletismografia. Resultados: foram estudados 429 pacientes, 172 normais e 275 com distúrbio ventilatório obstrutivo. Os pacientes normais apresentaram uma variação média na CRF de 120ml, no VR de 115ml e na CI de 50ml com BD (p<0,05). Os pacientes com DVO apresentaram variação média na CRF de 263ml, no VR de 271ml e na CI de 185ml (p<0,05). Entre os pacientes DVO com RBD pela espirometria encontrou-se variação média na CRF de 387ml, no VR de 287ml e na CI de 295ml (p<0,001). Entre os pacientes DVO NRBD pela espirometria, a CRF variou 180ml, o VR 140ml e a CI 111ml (p<0,004). Conclusões: pacientes DVO RBD pelo VEF1, tiveram variações médias da CRF, do VR e da CI superiores ao limite superior do intervalo de confiança de 95% (IC 95%) da variação dos volumes dos pacientes normais. Entretanto, nos pacientes com DVO NRBD pelo VEF1 apenas a variação média da CI ultrapassou o limite superior do IC95% dos pacientes normais (100ml), indicando que apesar de não haver resposta ao BD pelo VEF1 (critério de fluxo), houve resposta ao BD pela CI (critério de volume).

Introduction: the response to brochodilators (BD) measured on the forced expiratory volume on the 1 second (FEV1) is already universally established, but the variations of pulmonary volumes as responsiveness criteria to BD need to be better assessed. Methods: patients were divided in two groups (obstructive ventilatory disturb OVD and normal) on the spirometry and were classified according to the 1996's Brazilian Guidelines. These groups were subdivided in responsive to the BD (RBD) and non responsive (NRBD). All patients were submitted to pulmonary volumes measurements (residual volume-RV, functional residual volume-FRC, and inspiratory capacity IC) on the plethysmography. Results: we studied 429 patients, 172 normal and 257 with obstructive disorder. The normal patients presented a medium variation of 120ml on the FRC, 115ml on the RV, and 50ml on the IC with the BD (p<0,05). The obstructive patients presented a medium variation of 263ml on the FRC, 271ml on the RV, and 185ml on the IC (p<0,05). Among the patients with OVD-RBD by the spirometry was found a medium variation of 180ml on the FRC, 140ml on the RV, and 111ml on th IC (p<0,004). Conclusion: patients with obstructive ventilatory disorder RBD on the FEV1 had superior variations of the FRC, RV and IC compared to the volumes of normal patients. However in patients with OVD NRBD on the FEV1 only the IC exceeded the superior limit of normal patients (100ml), indicating that even though there was no response to the BD on the FEV1 (flow criteria), there was response to the BD on the IC (volume criteria)

Hipertensäo arterial pulmonar em paciente com hipotireoidismo em uso de anticoncepcional oral / Pulmonary arterial hypertension in one patient with hypothyroidism and on oral contraceptive

É apresentado um caso de mulher branca, de 34 anos de idade em tratamento para hipotireoidismo há oito anos, que desenvolveu hipertensäo arterial pulmonar há cerca de seis meses, após o início de uso de anticoncepcional oral. A evoluçäo foi favorável, com suspensäo do progestágeno, administraçäo de bloqueador de canais de cálcio, diurético e anticoagulante oral. Há relatos de disfunçöes da tireóide e uso de contraceptivo hormonal serem fatores de risco de desencadeamento de hipertensäo arterial pulmonar.

Relación entre el bajo peso de los recién nacidos y las edades de las madres, en los nacimientod registrados en la Maternidad Nacional del Hospital de Clínicas, desde enero de 1997 hasta diciembre de 1999

Estudio analítico observacional de una población enfocada a mujeres de 10 a 35 años, para determinar la relación existente entre el bajo peso de los recién nacidos y las madres adolescentes. Presenta las complicaciones obstétricas y fetales, además de las implicancias de los factores de riesgo

Opinión de Profesores Instructores y alumnos de la Escuela de Enfermería del Instituto Dr. Andrés Barbero de la Universidad Nacional de Asunción con relación a las actividades que se realizan en la práctica profesional

Estudio descriptivo, que presenta la importancia de la práctica profesional, tanto para docentes como estudiantes de la Escuela de Enfermería de la UNA. Ofrece el perfil profesional del Licenciado/a en Enfermería y describe factores de la Política de las actividades en la práctica profesional; relacionados con la enseñanza