ABSTRACT Objective: The aim of this double-blind, placebo-controlled and parallel- arm randomized clinical trial was to evaluate the effects of Lactobacillus rhamnosus SP1-containing probiotic sachet and azithromycin tablets as an adjunct to nonsurgical therapy in clinical parameters and in presence and levels of Tannerella forsythia, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans. Material and Methods: Forty-seven systemically healthy volunteers with chronic periodontitis were recruited and monitored clinically and microbiologically at baseline for 3, 6 and 9 months after therapy. Subgingival plaque samples were collected from four periodontal sites with clinical attachment level ≥1 mm, probing pocket depth ≥4 mm and bleeding on probing, one site in each quadrant. Samples were cultivated and processed using the PCR technique. Patients received nonsurgical therapy including scaling and root planing (SRP) and were randomly assigned to a probiotic (n=16), antibiotic (n = 16) or placebo (n = 15) group. L. rhamnosus SP1 was taken once a day for 3 months. Azithromycin 500mg was taken once a day for 5 days. Results: All groups showed improvements in clinical and microbiological parameters at all time points evaluated. Probiotic and antibiotic groups showed greater reductions in cultivable microbiota compared with baseline. The placebo group showed greater reduction in number of subjects with P. gingivalis compared with baseline. However, there were no significant differences between groups. Conclusions: The adjunctive use of L. rhamnosus SP1 sachets and azithromycin during initial therapy resulted in similar clinical and microbiological improvements compared with the placebo group.

RESUMO Os fungos podem se adaptar a condições ambientais e produzir diferentes respostas fisiológicas. O objetivo deste estudo foi verificar a existência de ecótipos de temperatura para Sclerotinia sclerotiorum em isolados do Brasil e dos EUA. Dez isolados de S. sclerotiorum de regiões tropicais e subtropicais do Brasil e seis dos EUA foram avaliados para crescimento micelial, patogenicidade, agressividade em feijão, canola e soja e formação de apotécio em temperaturas contrastantes. Para o crescimento micelial, independente da origem do isolado, todos cresceram mais rápido a 20oC em comparação com 27oC. Para patogenicidade e agressividade, a severidade da doença foi maior a 20oC do que a 30oC, para todos isolados. Para produção de apotécio, somente isolados do Brasil foram capazes de produzir apotécio sem pré-condicionamento. Com pré-condicionamento a 4oC por 40 dias, os isolados do Brasil e dos EUA produziram apotécio. Nenhum dos 16 isolados foi capaz de produzir apotécio a 30oC, após 40 dias. Os resultados não mostraram uma adaptação de isolados de S. sclerotiorum do Brasil para crescer ou colonizar folíolos em temperaturas mais altas, em comparação com os isolados dos EUA. Apenas escleródios de isolados de S. sclerotiorum do Brasil foram capazes de germinar carpogenicamente sem pré-condicionamento.

ABSTRACT Fungi can adapt to environmental conditions and produce different physiological responses. The aim of this study was to verify the existence of Sclerotinia sclerotiorum temperature ecotypes in isolates from Brazil and the USA. Ten S. sclerotiorum isolates from tropical and subtropical regions of Brazil and six isolates from the USA were used to measure mycelial growth, pathogenicity and aggressiveness on bean, canola and soybean, as well as apothecial formation at contrasting temperatures. For mycelial growth, regardless of the origin, all isolates grew faster at 20°C, compared to 27°C. For pathogenicity and aggressiveness, disease severity was greater at 20°C than at 30°C considering all isolates. As regards apothecial production, only Brazilian isolates were capable of producing apothecia with no preconditioning. After preconditioning at 4°C during 40 days, isolates from Brazil and the USA produced apothecia. None of the 16 isolates was capable of producing apothecia at 30oC after 40 days. Results indicated no adaptation of S. sclerotiorum isolates from Brazil to grow or colonize leaflets at higher temperatures, compared to isolates from the USA. Only sclerotia from S. sclerotiorum isolates from Brazil were capable of germinating carpogenically without preconditioning.

Congenital hyperinsulinism (CH) is the most frequent cause of persistent hypoglycemia in the newborn and it is characterized by an inappropriately elevated insulin level in presence of hypoglycemia. Initial management is medical treatment, but if it fails, partial pancreatectomy is the surgical procedure of choice. Objective: To report a case of a newborn with CH to aware on this condition and update its multidisciplinary management. Case report: The case of a full term newborn with severe symptomatic hypoglycemia unresponsive to medical treatment is reported . Ultrasonography and abdominal MRI were normal, Gallium 68 PET/CT showed uptake in the body of the pancreas. Laparoscopic corporocaudal pancreatectomy was performed. The patient resolved hypoglycemia and was discharged without incidents. Genetic study was consistent with focal CH, confirmed later by biopsy. Discussion: This case report exposes the clinical presentation of CH and its diagnosis. It emphasizes on the surgical treatment and the advantages of minimally invasive technique.

El hiperinsulinismo congénito (HC) es la causa más frecuente de hipoglicemia persistente del recién nacido (RN), y se caracteriza por secreción no regulada de insulina frente a hipoglicemia. El manejo inicial es médico, pero en caso de fracaso, la pancreatectomía parcial es el tratamiento quirúrgico de elección. Objetivo: Comunicar el caso de un RN con HC con énfasis en el estudio genético, de imágenes y manejo quirúrgico, incluyendo la cirugía laparoscópica. Caso clínico: RN de término, con hipoglicemia prolongada por HC que no respondió al tratamiento médico. La ultrasonografía y resonancia nuclear magnética abdominal fueron normales. PET/CT Galio 68 evidenció aumento de metabolismo glucídico focalizado en el cuerpo pancreático. Se realizó pancreatectomía córporo-caudal laparoscópica sin incidentes, con mejoría de la hipoglicemia. El estudio genético fue compatible con HC focal, confirmado luego por biopsia. Conclusión: Se expone acerca de las características clínicas y diagnóstico del HC, en particular el rol de la cirugía y ventajas del abordaje laparoscópico.

Objective The aim of this double- blind, placebo- controlled parallel- arm, randomised clinical trial was to evaluate the clinical effects of a sachet of Lactobacillus rhamnosus SP1-containing probiotic as an adjunct to non-surgical therapy. Material and method Fourty- nine subjects were screened for their elegibility to participate in this study. Twenty-eight systemically healthy volunteers with chronic periodontitis were enrolled and monitored clinically at baseline, and at 3 and 6 months after therapy. Clinical parameters measured included plaque index, bleeding on probing, pocket probing depths (PPD), and clinical attachment loss. Patients received non-surgical therapy including scaling and root planing (SRP), and were randomly assigned to a test (SRP + probiotic, n = 14) or control (SRP + placebo, n = 14) group. The administration of a sachet of Lactobacillus rhamnosus SP1 probiotic once a day for 3 months commenced after the last session of SRP. Results Both test and control groups showed improvements in clinical parameters at all time points evaluated. Furthermore, at initial visits and after 6 months follow-up, the test group showed a statistically significant reduction in percentage of sites, teeth, and number of participants with PPD ≥ 5 mm. Conclusions The results of this trial indicate that oral administration of L. rhamnosus SP1 sachets during initial therapy resulted in similar clinical improvements compared to SRP alone.

Objetivo El objetivo de este ensayo clínico aleatorizado, doble ciego, de brazos paralelos y controlado por placebo fue evaluar el efecto clínico del consumo de Lactobacillus rhamnosus SP1 en un polvo de disolución oral, adicional a la terapia periodontal no quirúrgica. Material y método Cuarenta y nueve sujetos fueron examinados para participar en este estudio. Veintiocho participantes sistémicamente sanos, con diagnóstico de periodontitis crónica, fueron reclutados y monitorizados clínicamente en el tiempo basal, 3 y 6 meses después de la terapia periodontal. Los parámetros clínicos registrados fueron presencia de placa, sangrado al sondaje, profundidad al sondaje y pérdida de inserción clínica. Todos recibieron terapia periodontal no quirúrgica, incluyendo pulido y alisado radicular (PAR), y fueron asignados aleatoriamente al grupo experimental (PAR + probiótico, n = 14) o control (PAR + placebo, n = 14). Luego de la última sesión de PAR debieron ingerir un sobre con polvo de disolución oral de Lactobacillus rhamnosus SP1 o placebo, una vez al día durante 3 meses. Resultados Ambos grupos mejoraron sus parámetros clínicos en todos los tiempos evaluados. Además, el grupo experimental redujo significativamente el porcentaje de sitios, dientes y número de participantes con profundidad al sondaje ≥ 5 mm entre el tiempo basal y los 6 meses postratamiento. Conclusiones La administración oral de L. rhamnosus SP1 asociado a la terapia periodontal genera similares mejorías en los parámetros clínicos comparado con solo usar terapia periodontal en el tratamiento de la periodontitis crónica en adultos.

Durante as safras 2008/09, 2009/10 e 2010/11 foi monitorada a sensibilidade do fungo Phakopsora pachyrhizi aos fungicidas tebuconazol, ciproconazol, metconazol e protioconazol (inibidores da desmetilação, IDMs). Folhas destacadas de soja foram tratadas com os fungicidas em doses variando de zero a 32 mg L-1 (tebuconazol, ciproconazol e metconazol) e zero a 8 mg L-1 (protioconazol) e inoculadas com esporos de P. pachyrhizi provenientes de lavouras de soja de diferentes regiões produtoras. As folhas inoculadas foram incubadas em placas de Petri com papel umedecido, a 23°C ± 2ºC, e a severidade da ferrugem estimada 15 dias após a inoculação. Foram determinadas as doses efetivas para reduzir 50% da severidade da doença (DE50). O fungicida protioconazol apresentou a maior atividade intrínseca, com valores de DE50 variando de 0,000001 mg L-1 a 0,39 mg L-1. Os valores de DE50variaram de 0,001 mg L-1 a 1,49 mg L-1 para tebuconazol; 0,001 mg L-1 a 3,27 mg L-1 para ciproconazol e 0,004 mg L-1 a 3,89 mg L-1 para metconazol. As medianas de DE50 para todos os fungicidas avaliados foram inferiores a 0,5 mg L-1, nas três safras. As correlações (r) entre as DE50 dos quatro fungicidas foram significativas (p<0,05), mostrando a existência de resistência cruzada entre os fungicidas. A variação nos valores de DE50 no monitoramento mostra uma coexistência de populações com diferentes níveis de sensibilidade aos IDMs no campo.

During 2008/09, 2009/10 and 2010/11 growing seasons, the sensitivity of the fungus Phakopsora pachyrhizi to the fungicides tebuconazole, cyproconazole metconazole and prothioconazole (demethylation inhibitors, DMIs) was monitored. Detached soybean leaves were treated with those fungicides at concentrations varying from zero to 32 mg L-1 (tebuconazole, cyproconazole and metconazole) and from zero to 8 mg L-1(prothioconazole) and were inoculated with P. pachyrhizi spores collected from different Brazilian soybean producing regions. Inoculated leaves were incubated in Petri plates containing moistened paper towel at 23° C ± 2° C, and the disease severity was estimated at 15 days after inoculation. The effective concentrations to reduce the disease severity by 50% (EC50) were estimated. The fungicide prothioconazole presented the highest intrinsic activity and its EC50 ranged from 0.000001 mg L-1 to 0.39 mg L-1. EC50 values ranged from 0.001 mg L-1 to 1.49 mg L-1 for tebuconazole, 0.001 mg L-1 to 3.27 mg L-1 for cyproconazole and 0.004 mg L-1 to 3.89 mg L-1 for metconazole. EC50medians of all tested fungicides were lower than 0.5 mg L-1 in the three growing seasons. The correlations (r) among the EC50 values of the four fungicides were significant (p<0.05), indicating cross-resistance among the products. The EC50 variation in the monitoring shows coexistence of populations with different levels of sensitivity to DMIs in the field.

A estimativa visual da severidade de doenças em plantas nem sempre se correlaciona com o efeito desta sobre a atividade fotossintética do hospedeiro. O objetivo do trabalho foi avaliar a interferência dos fungos Corynespora cassiicola e Erysiphe diffusa, causadores da mancha-alvo e do oídio na cultura da soja, respectivamente, na eficiência fotossintética de folhas infectadas. A fotossíntese foi relacionada com a área foliar doente por meio da equação Px/ Po=(1-x)β. Os parâmetros β (± erro padrão) estimados foram 2,78 (± 0,28) (p<0,05) para as folhas infectadas com C. cassiicola e 0,72 (±0,09) (p<0,05) e 0,77 (±0,15) (p=0,15) com E. diffusa, na primeira e segunda repetições. O valor obtido para C. cassiicola (β>1) indica que houve redução da eficiência fotossintética no tecido lesionado e em parte do tecido verde remanescente, enquanto que os valores obtidos para E. diffusa (β≤1) indicam que a estimativa visual da severidade da doença é um bom indicador do efeito do fungo na taxa fotossintética do hospedeiro.

The visual estimate of the disease severity in plants does not always correlate with its effect on the host's photosynthetic activity. The aim of this study was evaluate the interference of the fungi Corynespora cassiicola and Erysiphe diffusa, causal agents of target spot and powdery mildew on soybeans, in the photosynthetic efficiency of infected leaves. Photosynthesis was related to the diseased leaf area by the equation Px/Po = (1-x)β. The estimated parameters β (± standard error) were 2.78 (± 0.28) (p<0.05) for the leaves infected with C. cassiicola and 0.72 (± 0.09) (p<0.05) and 0.77 (± 0.15) (p=0.15) for those infected with E. diffusa in the first and the second replicates. The value obtained for C. cassiicola (β>1) indicates that there was a reduction in the photosynthetic efficiency in the injured tissue and in part of the remaining green tissue, while the values obtained for E. diffusa (β≤1) indicate that the visual estimate of the disease severity is a good indicator of the effect of the fungus on the photosynthetic rate.

O objetivo deste trabalho foi avaliar a diversidade genética entre isolados de Sclerotinia sclerotiorum do Brasil e dos EUA, determinar sua variabilidade quanto à agressividade e verificar a existência de interação isolado-cultivar. A variabilidade dos isolados foi determinada por agrupamento de compatibilidade micelial (ACM), e a agressividade dos isolados, por meio de inoculações, com corte da haste, em cultivares de soja. Dois experimentos de ACM e dois de agressividade foram realizados, com dois conjuntos de isolados. O primeiro conjunto incluiu nove isolados, do mesmo campo de soja no Brasil, e nove da região Meio-Oeste dos EUA. O segundo conjunto incluiu 16 isolados de várias regiões do Brasil e um dos EUA. No primeiro conjunto, 18 isolados formaram 12 ACMs diferentes. No segundo conjunto, 81% dos isolados do Brasil formaram um único ACM. Nenhum ACM comum foi observado entre os isolados do Brasil e dos EUA. Os isolados apresentaram diferenças quanto à agressividade no primeiro conjunto de isolados, mas não no segundo. Embora a agressividade tenha diferido no primeiro conjunto, as cultivares de soja e os isolados não interagiram significativamente. A classificação das cultivares permaneceu a mesma, independentemente da diversidade genética, da diferença quanto à agressividade e da região ou país de origem do isolado. Resultados da seleção de cultivares de soja, realizada pelo método de inoculação de corte da haste nos EUA, podem ser utilizados como referência para pesquisadores no Brasil.

The objective of this work was to evaluate the genetic diversity among Sclerotinia sclerotiorum isolates from Brazil and the USA, assess their aggressiveness variability, and verify the existence of an isolate-cultivar interaction. Isolate variability was determined by mycelial compatibility grouping (MCG), and isolate aggressiveness by cut-stem inoculations of soybean cultivars. Two experiments for MCGs and two for aggressiveness were conducted with two sets of isolates. The first set included nine isolates from the same soybean field in Brazil and nine from the Midwest region of the USA. The second set included 16 isolates from several regions of Brazil and one from the USA. In the first set, 18 isolates formed 12 different MCGs. In the second set, 81% of the isolates from Brazil grouped into a single MCG. No common MCGs were observed among isolates from Brazil and the USA. The isolates showed aggressiveness differences in the first set, but not in the second. Although aggressiveness differed in the first set, soybean cultivars and isolates did not interact significantly. Cultivar rank remained the same, regardless of the genetic diversity, aggressiveness difference, and region or country of origin of the isolate. Results from screening of soybean cultivars, performed by the cut-stem method in the USA, can be used as reference for researchers in Brazil.

The incidence of target spot disease on soybean has increased in recent years in Brazil even with intensive use of fungicides, and fungal resistance has been reported in recent studies. The objective of this study was to determine the fungicide sensitivity to carbendazim and prothioconazole in a sample of 24 isolates of Corynespora cassiicola from soybean collected from 1996 to 2011 in the states of Paraná, Mato Grosso and São Paulo ( Brazil) and Corpus Christi (Paraguay). The 50% effective concentration (EC50) values were estimated by the relative mycelium growth reduction on fungicide-amended medium with the doses of 0, 0.5, 1, 10, 100 and 1000 µg of active ingredient/ mL. For carbendazim, four highly resistant isolates (EC50>50 µg/mL) were observed from samples collected from Mato Grosso in 2008 and from Paraná and Mato Grosso in 2011. The EC50 values for prothioconazole ranged from 0.47 µg/mL to 26.44 µg/mL (mean: 5.02 µg/ ml). The results reinforce the occurrence of C. cassiicola resistance to benzimidazole in Paraná and Mato Grosso states.

Objetivou-se detectar a influência do estádio fenológico e da idade da folha de soja na infecção por Phakopsora pachyrhizi, agente causal da ferrugemasiática (FA). Plantas das cultivares BRS 154 e BRS 258 foram inoculadas, com suspensão de 10(5) urediniósporos/mL, nos estádios fenológicos V3, R1 e R5. Após 24 horas de câmara úmida, as plantas foram acondicionadas em condições de casa de vegetação por 20 dias. Avaliou-se o período latente médio (PLM) e a severidade. Para a avaliação da suscetibilidade de trifólios à FA utilizou-se a cultivar BRS 154 (V5). A inoculação foi realizada nos quatro primeiros trifólios. Aos 15 dias após a inoculação, os folíolos foram avaliados quanto à severidade, tamanho médio de lesão e freqüência de infecção. O estádio das plantas de soja não influenciou no PLM. As cultivares BRS 154 e BRS 258 tiveram PLM de 8 e 9 dias, respectivamente. As cultivares não se diferenciaram quanto à severidade da doença. Não houve diferença de severidade nos estádios V3 e R1, porém, os valores de severidade nesses estádios foram superiores ao valor de severidade no estádio R5, na avaliação realizada 8 dias após a inoculação. Em relação à suscetibilidade de folhas, o trifólio mais velho apresentou maiores valores de doença.

This work was conducted to study the influence of soybean growth stage and leaf age on the infection of Phakopsora pachyrhizi, the soybean rust pathogen. Soybean plants (cv. BRS 154 and BRS 258) at the V3, R1 and R5 growth stages were inoculated with a 1 x 10(5) urediniospores per mL suspension. After a period of 24 hours in dew chambers, all plants were removed from the chambers and placed under greenhouse conditions for 20 days. Mean latent period (PLM) and disease severity were estimated. The susceptibility of trifoliate leaves to soybean rust was estimated on cv. BRS 154 at the growth stage R5. Pathogen inoculation was done at the first four trifoliate leaves. Fifteen days after inoculation, leaflets of each trefoil were evaluated for disease severity, lesion mean size and infection frequency. Plants' growth stage did not influence the PLM. Cultivars BRS 154 and BRS 258 presented PLM of 8 and 9 days, respectively. There was no difference in disease severity at the growth stages V3 and R1, but those values were higher than at the R5 growth stage, 8 days after inoculation. The oldest trefoil showed the highest disease values.

One hundred and sixty seven fecal samples and three complete digestive tracts obtained from these penguins were analyzed using different parasitological methodologies. The presence ofTetrabothrius spp. eggs (Cestoda: Pseudophyllida), and the observation of Sporozoa oocists and Nematoda eggs both of undetermined species in the fecal samples is reported. In the same way a large number of nematodes identified as Streptocara spp. (Nematoda: Acuariidae) in different evolutionary stages in stomach, and the recovery of a whole 81 cm. Tetrabothrius spp. specimen (without the scolex) and one inmature tapeworm (Cestoda: Cyclophyllida) with unarmed rostellum in guts of Adéliepenguins chiks found dead in a protected zone of the Antarctic territory is reported.

Usando diferentes técnicas parasicológicas, fueron analizadas 167 muestras de heces y 3 tractos digestivos completos obtenidos de pingüinos Adelia {Pygoscelis adeliae) de una zona Antartica protegida. Se reporta la presencia de: huevos y una estróbila de Tetrabothrius spp. (Cestoda: Pseudophyllida); una tenia con un rostellum inerme (Cestoda: Cyclophyllida) en estado inmaduro; ooquistes de Esporozoos y huevos de Nematodos sin identificación; una gran cantidad de Streptocara spp. (Nematoda: Acuariidae) en diferentes estados de evolución.

A ferrugem da soja destacou-se nas últimas safras devido à alta severidade que vem causando nas lavouras de soja. Diversos estudos estão em andamento, para buscar informações sobre a resistência genética dos cultivares atualmente plantados. O objetivo do trabalho foi estudar parâmetros monocíclicos e o progresso da ferrugem da soja em diferentes genótipos e posições da copa, em casa-de-vegetação. Foram utilizados 7 cultivares (Uirapuru e BRS 134 Pintado, BRS 154, BRS 215, FT 2, BRS 231) e uma PI 459025 com gene de resistência Rpp4, inoculados com suspensão de esporos de Phakopsora pachyrhizi. Foram estudados para cada genótipo os períodos de incubação e latente. A avaliação de incidência da ferrugem foi realizada na planta toda e a da severidade a partir do aparecimento dos sintomas, a cada cinco dias, até o declínio das plantas em três posições na copa. Os valores foram transformados em área abaixo da curva de progresso da incidência (AACPI) e da severidade (AACPS) da doença. O período de incubação foi de seis dias para todos genótipos avaliados. Entretanto, o período latente variou de 6 a 12 dias. Houve diferença significativa entre os cultivares para AACPI. Os cultivares BRS 134, FT 2 e BRS 231 apresentaram maior valor de AACPI, diferenciando dos demais cultivares. Entre os cultivares com menor valor de AACPI destacou-se a PI 459025. Variação na intensidade da doença nos 8 genótipos avaliados, em relação à posição da copa, só pode ser observada para o terço médio da planta.

Soybean rust caused by the fungus Phakopsora pachyrhizi, has been known to drastically reduce yields. Diverse studies are now in progress, to search information on the genetic resistance of cultivar currently planted. The objective of this study was to evaluate the both incubation and latent period and the progress of soybean rust on several genotypes and three canopy position in greenhouse. The cultivars used were Uirapuru, BRS 134 Pintado, BRS 154, BRS 215, BRS 231, FT 2 and PI 459025 with a Ppp4 gene of resistance for soybean rust. Inoculations were performed by spraying inoculum suspension on the leaves. Disease incidence and severity were assessed every five days after the first disease symptoms were observed until plants were dead. The incubation period was six days for all genotypes evaluated. However, the latent period varied from six to twelve days. The incidence and severity values were transformed in proportion as the area under the disease incidence curve (AUDIC) and area under the disease severity curve (AUDSC). The cultivars BRS 134 Pintado, FT 2 and BRS 231 showed high values for AUDIC and were significant different from the others. PI 459025 showed the lowest value for AUDIC. AUDSC for PI 459025 cultivar was the lowest and FT 2 showed the highest value. Difference from disease intensity on eight genotypes, in function of the canopy position, only was observe to plant medium part.

This study try to know the teacher’s perception about the programs of early stimulation, researching on the areas of work, the fortresses and limitations that they find in their daily work with the children. To get the information, a field work was done interviewing 14 teachers of kinder garden (publics and privates). An evaluation of context was done using a constructed interview. The principal results were: 100% works as a priority the area of communication and language; 64.30% indicates that all the areas of the early stimulation are important; 71.43% does not rely on the right material and 50% on the suitable infrastructure. Likewise 50% of the interviewed ones think that the parents are not compromised in the work by the children.

Esta investigación busca conocer la percepción que tiene los maestros sobre los programa de estimulación temprana, indagando sobre las áreas de trabajo, las fortalezas y limitaciones que encuentran ellos en su trabajo diario con los niños. Para la recogida de información se hizo un trabajo de campo entrevistando a 14 profesoras de educación inicial y de nidos (nacionales y particulares). Se hizo una evaluación de contexto a través de una entrevista estructurada. Los principales resultados fueron: el 100% trabaja prioritariamente el área de comunicación y lenguaje; el 64.30% señala que todas las áreas de la estimulación temprana son importantes; el 71.43% no cuenta con el material adecuado y el 50% con la infraestructura adecuada. Asimismo, el 50% de las entrevistadas consideran que los padres no están comprometidos en el trabajo con los niños.