Resumo O conteúdo de dopamina nas estruturas cerebrais tem sido relacionado à excitabilidade neuronal e várias abordagens têm sido utilizadas para estudar este fenômeno durante o período de vulnerabilidade às crises. No presente trabalho, descrevemos os efeitos da depleção de dopamina após a administração de 6-hidroxidopamina (6-OHDA) na região pars compacta da substância negra de ratos submetidos ao modelo de epilepsia com pilocarpina. A susceptibilidade ao estado de mal epiléptico induzido pela pilocarpina, bem como a frequência de crises espontâneas e recorrentes durante o período crônico do modelo foi determinada. Sendo o hipocampo uma das principais estruturas afetadas no desenvolvimento desse modelo experimental de epilepsia, os níveis de dopamina nessa região foram determinados após a administração da droga. No primeiro experimento, 62% (15/24) dos ratos pré-tratados com 6-OHDA e 45% (11/24) daqueles que receberam ácido ascórbico como solução controle evoluíram para crises límbicas motoras e para o estado de mal epiléptico, após a administração de pilocarpina. A gravidade das crises durante o período agudo do modelo foi significativamente maior nos ratos epilépticos experimentais (56,52%) do que nos ratos controle (4,16%). No segundo experimento, não houve diferença significante entre os grupos quanto à frequência de crises na fase crônica do modelo. Nossos dados mostraram que a dopamina pode desempenhar um papel importante na gravidade das crises na fase aguda da pilo, o que parece ser exercido por sua ação inibitória da dopamina sobre a expressão motora das crises.

Abstract The dopamine content in cerebral structures has been related to neuronal excitability and several approaches have been used to study this phenomenon during seizure vulnerability period. In the present work, we describe the effects of dopamine depletion after the administration of 6-hidroxidopamine (6-OHDA) into the substantia nigra pars compacta of male rats submitted to the pilocarpine model of epilepsy. Susceptibility to pilocarpine-induced status epilepticus (SE), as well as spontaneous and recurrent seizures (SRSs) frequency during the chronic period of the model were determined. Since the hippocampus is one of main structures in the development of this experimental model of epilepsy, the dopamine levels in this region were also determined after drug administration. In the first experiment, 62% (15/24) of 6-OHDA pre-treated rats and 45% (11/24) of those receiving ascorbic acid as control solution progressed to motor limbic seizures evolving to SE, after the administration of pilocarpine. Severeness of seizures during the model´s the acute period, was significantly higher in epileptic experimental rats (56.52%), than in controls (4.16%). In the second experiment, the frequency of seizures in the model’s chronic phase did not significantly change between groups. Our data show that dopamine may play an important role on seizure severity in the pilo’s model acute period, which seems to be due to dopamine inhibitory action on motor expression of seizure.

Reuse made by the separation of whey can reduce the waste of human milk. However, the authors are not aware of the literature on treatments in human whey, made available by pasteurization holder, freeze-drying, spray drying, and high hydrostatic pressure. In this context, the present study applied treatments to human whey and evaluated their lipid and immunological quality. Among the results, a different formation in the triacylglycerol profile was evidenced after the application of spray drying and high hydrostatic pressure, while pasteurization and freeze-drying presented ion intensity close to the control human whey. In addition, pasteurization proved to be adequate for lipid nutritional quality and transformation factor-β1 (TGF-β1) concentration, while an increase in interleukin-10 (IL-10) levels was promoted, between 73 and 80%, after freeze-drying, spray drying, and high hydrostatic pressure. Through the principal component analysis, it is noteworthy that the processes presented divergences in terms of the effects caused, with similarity only between pasteurization and freeze-drying in the composition of fatty acids. However, it was observed that all processes were able to maintain the nutrients. Nevertheless, it is relevant to consider individual characteristics presented and the interest in the desired quality, which can be promising as a complementary product to infant feeding.

ABSTRACT Objective: To report outcomes from the largest multicenter series of penile cancer patients undergoing video endoscopic inguinal lymphadenectomy (VEIL). Materials and Methods: Retrospective multicenter analysis. Authors of 21 centers from the Penile Cancer Collaborative Coalition-Latin America (PeC-LA) were included. All centers performed the procedure following the same previously described standardized technique. Inclusion criteria included penile cancer patients with no palpable lymph nodes and intermediate/high-risk disease and those with non-fixed palpable lymph nodes less than 4 cm in diameter. Categorical variables are shown as percentages and frequencies whereas continuous variables as mean and range. Results: From 2006 to 2020, 210 VEIL procedures were performed in 105 patients. Mean age was 58 (45-68) years old. Mean operative time was 90 minutes (60-120). Mean lymph node yield was 10 nodes (6-16). Complication rate was 15.7%, including severe complications in 1.9% of procedures. Lymphatic and skin complications were noted in 8.6 and 4.8% of patients, respectively. Histopathological analysis revealed lymph node involvement in 26.7% of patients with non-palpable nodes. Inguinal recurrence was observed in 2.8% of patients. 10y- overall survival was 74.2% and 10-y cancer specific survival was 84.8%. CSS for pN0, pN1, pN2 and pN3 were 100%, 82.4%, 72.7% and 9.1%, respectively. Conclusion: VEIL seems to offer appropriate long term oncological control with minimal morbidity. In the absence of non-invasive stratification measures such as dynamic sentinel node biopsy, VEIL emerged as the alternative for the management of non-bulky lymph nodes in penile cancer.

ABSTRACT Mangrove ecosystems are sensitive to oil, as spills can impair developmental processes of mangrove vegetation. Since the 2010s, the Brazilian equatorial margin, more specifically the Pará-Maranhão Basin and the mouth of the Amazonas River, has been affected by oil runoff from urban activities and the increased risk from exploratory deepwater drilling for oil extraction. Dispersal of mangrove propagules occurs during the tidal cycles, when the presence of tensors in the water can affect germination. We analyzed the effects of diesel oil on the germination of propagules of the two most common mangrove species in the region, Laguncularia racemosa and Avicennia germinans, in six treatments of diesel oil in the water (0.5%, 1%, 1.5%, 2%, 3% and 4%) and a diesel-free control. The response variables were germinability (G%), mean germination time, mean germination speed and the germination speed index (GSI). G% and GSI in L. racemosa propagules differed significantly between the control and all treatments (G% and GSI < 10% in the 3% treatment). Propagules of A. germinans were more resistant, and the physiological variables did not differ significantly among treatments and control (G% > 90% in all treatments). Our results indicate that, at the germination stage, L. racemosa was more susceptible than A. germinans to contamination by automotive diesel oil.

RESUMO Os ecossistemas de manguezais são sensíveis ao óleo, pois os derramamentos podem prejudicar os processos de desenvolvimento da vegetação de mangue. Desde a década de 2010, a margem equatorial brasileira, mais especificamente a Bacia do Pará-Maranhão e a foz do Rio Amazonas, vem sendo afetada pelo vazamento de petróleo proveniente de atividades urbanas e pelo aumento do risco de perfurações exploratórias em águas profundas para extração de petróleo. A dispersão dos propágulos do manguezal ocorre durante os ciclos das marés, quando a presença de tensores na água pode afetar a germinação. Analisamos os efeitos do óleo diesel na germinação de propágulos de duas espécies de mangue mais comuns na região, Laguncularia racemosa e Avicennia germinans, em seis tratamentos de óleo diesel na água (0,5%, 1%, 1,5%, 2%, 3% e 4%) e um controle sem óleo diesel. As variáveis resposta foram germinabilidade (G%), tempo médio de germinação, velocidade média de germinação e índice de velocidade de germinação (IVG). G% e IVG dos propágulos de L. racemosa diferiram significativamente entre o controle e todos os tratamentos (G% e GSI < 10% no tratamento 3%). Os propágulos de A. germinans foram mais resistentes e as variáveis fisiológicas não diferiram significativamente entre tratamentos e controle (G% > 90% em todos os tratamentos). Nossos resultados indicam que, no estágio de germinação, L. racemosa foi mais suscetivel que A. germinans à contaminação por óleo diesel automotivo.

Originária da América Tropical, a goiabeira ( Psidium guajava L. ) tem grande símbolo para o Brasil. Contudo, a evolução do patossistema designado como declínio da goiabeira, problema fitossanitário provocado pelo parasitismo das raízes pelo nematoide-das-galhas ( Meloidogyne enterolobii Yang &amp; Eisenback) em associação com fungos oportunistas, dizimou muitos pomares em todas as regiões do Brasil e em outros países. No presente estudo, propagou-se vegetativamente, por miniestaquia, acessos de goiabeiras seminíferas mantendo seus genótipos preservados e reavaliando a resistência por meio dos clones de forma a comprovar ou não as reações das plantas hospedeiras. Os resultados apontam para alta virulência do parasita, bem como alta hospedabilidade da espécieP. guajava , além da existência de grande variação da reação entre plantas do mesmo genótipo e entre genótipos distintos, o que indica que a estratégia de preservação do germoplasma e a reavaliação da reação em clones pode ser importante na busca e seleção de germoplasma com algum grau de ou resistência de tolerância a M. enterolobii . A progênie da cv. Paluma P02R5R2 obteve uma menor média de Fator de Reprodução do parasita (FR = 22,11) entre os genótipos avaliados, sendo classificado como moderadamente resistente e preservado para estudos posteriores.

Originária da América Tropical, a cultura da goiabeira (Psidium guajava L.) tem grande relevância para o Brasil. Contudo, surgimento do patossistema designado como declínio da goiabeira, problema fitossanitário provocado pelo parasitismo das raízes por nematoides da espécie Meloidogyne enterolobii Yang &amp; Eisenback em associação com fungos oportunistas, dizimou muitos pomares em todas as regiões do Brasil e em outros países. No presente estudo, propagou-se vegetativamente, por miniestaquia, acessos de goiabeiras seminíferas mantendo seus genótipos preservados e reavaliando a resistência por meio dos clones de forma a comprovar ou não as reações das plantas hospedeiras. Os resultados apontam para alta virulência do parasita, bem como alta hospedabilidade da espécie P. guajava, além da existência de grande variação da reação entre plantas do mesmo genótipo e entre genótipos distintos, o que indica que a estratégia de preservação do germoplasma e a reavaliação da reação em clones pode ser importante na busca e seleção de germoplasma com algum grau de resistência ou características de tolerância ao M. enterolobii. A progênie da cv. Paluma P02R5R2 obteve a menor média de Fator de Reprodução do parasita (FR = 22,11) entre os genótipos avaliados, sendo classificada como moderadamente resistente e preservada para estudos posteriores.

Resumen Introducción. El interés de la población por el consumo de productos frescos como las hortalizas ha ido en aumento a través de los años. Existen diferentes métodos de cultivo para producir hortalizas como zanahoria, tomate y repollo. Los sistemas de cultivo convencional y orgánico son los más comunes. Objetivo. Comparar la microbiología de zanahoria, tomate y repollo, provenientes de fincas con sistemas productivos de agricultura orgánica o convencional en Costa Rica. Materiales y métodos. El trabajo se realizó en Cartago, Heredia y Alajuela, Costa Rica, de setiembre 2020 a noviembre 2021, durante la época lluviosa en todos los casos. Se muestrearon hortalizas de fincas modelo de producción de zanahoria, tomate y repollo, una de producción orgánica y tres de producción convencional. Se determinaron recuentos totales de aerobios mesófilos, coliformes totales, Escherichia coli, mohos y levaduras, y la ausencia/presencia de Listeria monocytogenes. Resultados. Los recuentos totales aerobios, de mohos y levaduras y de coliformes totales de la zanahoria, el repollo y el tomate de cultivo convencional y orgánico no difirieron entre sí. En las hortalizas estudiadas los recuentos de E. coli fueron de <1 log UFC/g y hubo ausencia de Listeria monocytogenes. Se encontró presencia de Listeria sp. en las muestras de zanahoria de cultivo convencional, en contraste con su ausencia en la zanahoria de cultivo orgánico. Conclusiones. L. monocytogenes estuvo ausente en las hortalizas estudiadas, que en general, tuvieron cargas microbianas aceptables según la normativa y similares a estudios reportados a nivel mundial. Bajo condiciones controladas, si se comparan con aquellas obtenidas de sistemas de producción convencional, es posible obtener hortalizas con la técnica de cultivo orgánico sin detrimento en su calidad microbiológica y con los beneficios derivados para la salud del consumidor y el ambiente.

Abstract Introduction. Interest in consuming fresh products such as vegetables has been increasing over the years. Different cultivation methods to produce vegetables such as carrots, tomatoes, and cabbages are available. Conventional and organic farming systems are the most common. Objective. To compare the microbiology of carrot, tomato and cabbage from farms with organic or conventional agricultural production systems in Costa Rica. Materials and methods. Sampling was carried out in Cartago, Heredia and Alajuela, Costa Rica, from September 2020 to November 2021, during the rainy season in all cases. Vegetables from model farms of carrot, tomato and cabbage, one of organic production and three of conventional production, were sampled. Microbial counts of total aerobic mesophilic, total coliforms, Escherichia coli, yeasts and molds, and the absence/presence of Listeria monocytogenes were determined. Results. Total aerobic, yeast and mold and total coliform counts for conventionally and organically grown carrots, cabbages, and tomatoes did not differ from each other. In the vegetables studied, E. coli counts were <1 log CFU/g and Listeria monocytogenes was absent. Presence of Listeria sp. in carrot samples from conventional cultivation was found in contrast to its absence in carrots from organic cultivation. Conclusions. L. monocytogenes was absent in the vegetables studied, which in general had acceptable microbial loads according to regulations and similar to studies reported worldwide. Under controlled conditions, when compared to those obtained from conventional production systems, it is possible to obtain vegetables from organic farming without detriment to their microbiological quality and with the derived benefits for the health of the consumer and the environment.

ABSTRACT Cowpea is a legume that is grown worldwide and used for different purposes, especially as green grains. However, considering the low availability of cowpea cultivars for green grain production, selecting genotypes that have better traits for this purpose is necessary. In this context, the objective of this study was to estimate genetic parameters and evaluate different selection indices for identifying superior cowpea genotypes and subsidizing cowpea breeding programs focused on green grain production. A field experiment was conducted at the Center of Agricultural Sciences of the Federal University of Ceará (UFC), Ceará, Brazil. The treatments consisted of 42 cowpea genotypes from the Active Germplasm Bank of the UFC. Fourteen traits were used for characterization. The experiment was conducted in an augmented block design with four controls. The data obtained were subjected to analysis of variance, and genetic parameters, correlations, and selection indices were determined. The traits days to flowering (DFL), days to fruiting (DFR), green pod weight (GPW), green pod width (GW), green pod length (GPL), green pod thickness (GPT), number of grains per pod (NGP), and green grain thickness (GGT) showed heritability higher than 70%, indicating that selection in an early generation is favorable. The genetic correlations between the trait pairs DFL×DFR, GPW×GPL, and GW×GGT were higher than the phenotypic and environmental correlations. Genotypes CE-228, CE-688, CE-994, CE-165, CE-796, and BRS-Paraguaçu showed simultaneous superiority for the evaluated traits and are the most appropriate for green grain production.

RESUMO O feijão-caupi é uma das leguminosas mais cultivadas no mundo, sendo comercializado para diversas finalidades, como o feijão verde. Considerando a reduzida disponibilidade de cultivares de feijão-caupi para o mercado de feijão verde, é necessário selecionar genótipos que ofereçam características intrínsecas para este fim. Assim, objetivou-se estimar parâmetros genéticos e índice de seleção na identificação de genótipos superiores de feijão-caupi, fornecendo subsídios para programas de melhoramento visando a produção de grãos verdes. O experimento foi conduzido em campo, no Centro de Ciências Agrárias da Universidade Federal do Ceará (UFC), Ceará, Brasil. Os tratamentos consistiram de 42 genótipos de feijão-caupi do Banco Ativo de Germoplasma da UFC. Quatorze descritores foram utilizados para a caracterização. O experimento foi montado em um delineamento de blocos aumentados, com quatro testemunhas adicionais. Foi realizada análise de variância, parâmetros genéticos, correlação e índices de seleção. As características de dias para floração, dias para frutificação, peso da vagem verde, largura da vagem verde, comprimento da vagem verde, espessura da vagem verde, número de grãos por vagem e espessura do grão verde apresentaram valores de herdabilidade acima de 70%, indicando que a seleção em geração precoce com base nestas características é favorável. As correlações genéticas entre os pares de caracteres DFL×DFR, GPW×GPL, e GW×GGT foram superiores às correlações fenotípicas e ambientais. Os genótipos CE-228, CE-688, CE-994, CE-165, CE-796 e BRS-Paraguaçu apresentam superioridade simultânea para as características avaliadas e os mais adequados para o mercado de grãos e vagens verdes.

Abstract This study compared the preparation of long oval-shaped root canals using WaveOne Gold and XP-endo Shaper systems, both supplemented or not with manual instrumentation. Twenty-four long oval-shaped canals of mandibular incisors were distributed into two groups according to the instrumentation: WaveOne Gold Primary or XP-endo Shaper systems. All root canals were manually instrumented with a size 25 K-file after automated preparation. The specimens were scanned by using a micro-CT device (17.42 µm) before and after automated preparation and manual instrumentation. The increased surface of the root canal and the percentage of untouched areas were assessed. Both WaveOne Gold and XP-endo Shaper systems increased the root canal surface and had similar untouched areas (p>0.05). Supplementary instrumentation increased the surface of the root canal and decreased the untouched walls (p<0.05). WaveOne Gold and XP-endo Shaper systems provided a similar preparation of long oval-shaped canals and manual instrumentation improved their preparation.

Resumo Este estudo comparou o preparo de canais radiculares ovais longos usando os sistemas WaveOne Gold e XP-endo Shaper, ambos complementados ou não com instrumentação manual. Vinte e quatro canais longos e ovais de incisivos inferiores foram distribuídos em dois grupos de acordo com a instrumentação: sistemas WaveOne Gold Primary ou XP-endo Shaper. Todos os canais foram instrumentados manualmente com uma lima K tamanho 25 após preparo automatizado. Os espécimes foram escaneados usando um dispositivo de micro-CT (17,42 µm) antes e após o preparo automatizado e instrumentação manual. O aumento de superfície do canal radicular e a porcentagem de áreas intocadas foram avaliadas. Ambos os sistemas WaveOne Gold e XP-endo Shaper aumentaram a superfície do canal radicular e apresentaram áreas intocadas semelhantes (p>0,05). A instrumentação complementar aumentou a superfície do canal radicular e diminuiu o percentual de áreas intocadas (p <0,05). Os sistemas WaveOne Gold e XP-endo Shaper tiveram o preparo de canais ovais longos semelhantes, e a instrumentação manual melhorou o preparo.

Abstract Background Postdural puncture headache (PDPH) is a common complication of neuraxial techniques which delays patients’ discharge. Sphenopalatine ganglion block (SPGB) is a safe bedside technique with comparable efficacy to Epidural Blood Patch, the gold-standard treatment. There is no evidence on the ideal timing for SPGB performance. We aimed to evaluate the difference between early versus late SPGB concerning efficacy, symptom recurrence and hospital length of stay. Methods We present an observational study with 41 patients diagnosed with PDPH who were submitted to SPGB with ropivacaine 0,75%. The study sample (n = 41) was divided in two groups: an early (less than 24 hours after diagnosis) and a late (more than 24 hours after diagnosis) SPGB group. Pain was evaluated 15 minutes after the block and follow up occurred daily until patients were discharged. Patients’ demographic characteristics, neuraxial technique, timing of SPGB, qualitative pain relief and post-SPGB length of stay were registered and analyzed with SPSS statistics (v26) software. Results Early SPGB resulted in a significant reduction in length of stay (p = 0,009) and symptom recurrence (p = 0,036), showing equally effective pain relief, compared to late SPGB. Conclusions SPGB was equally effective in both groups. Data showed that early SPGB reduces length of hospital stay and symptom recurrence, which potentially allows early resumption of daily activities and a reduction in total health costs.

RESUMEN En tomate (Solanum lycopersicum L.), las plantas injertadas pueden conducirse a más de un eje, siendo de interés discriminar si las respuestas se deben al portainjerto o a la forma de conducción. Este trabajo tuvo como objetivo estudiar el efecto del injerto y el número de ejes por planta sobre su comportamiento fenológico y productivo. Se condujeron dos ensayos bajo un invernadero metálico parabólico de 24x40 m en La Plata, Buenos Aires, Argentina (33º56'42,6”S, 60º33'35,6”W), evaluando: 1) Planta: ‘SVTH 2900’ sin injertar, autoinjertado e injertado sobre ‘Multifort’; 2) Ejes por planta: uno, dos, cuatro. El diseño fue en bloques completos aleatorizados con arreglo factorial 2x3 con 3 repeticiones (2020) y 3x3 con 4 repeticiones (2021). Se registraron los días y grados-día acumulados desde trasplante, rendimiento y peso medio de frutos. Los datos fueron analizados usando un análisis de varianza y prueba de Tukey. No hubo diferencias en el momento de inicio de cosecha, y el ciclo productivo se prolongó por el injerto y el mayor número de ejes. El injerto incrementó el peso medio de frutos, el rendimiento total en 32 a 34% y la producción de frutos con peso >150 g en 45%. La conducción a dos ejes promovió el rendimiento total. Además, el número de frutos fue superior en plantas a dos ejes en 2020, mientras que aumentó en plantas injertadas conducidas a cuatro ejes en 2021. En las condiciones de ensayo, puede recomendarse el uso de plantas injertadas y la conducción a dos ejes, ya que se observa un mejor comportamiento fenológico y productivo.

ABSTRACT Grafted tomato (Solanum lycopersicum L.) plants can be pruned to more than one stem, and therefore knowing if the observed growth responses are triggered by rootstock or pruning method is a matter of interest. The aim of this work was to study the effect of grafting and number of stems per plant on its phenological and productive behavior. Two greenhouse trials were carried out under a 24x40 m metallic greenhouse with parabolic roof in La Plata, Buenos Aires, Argentina (33º56'42,6”S, 60º33'35,6”W), evaluating: 1) Plant: non-grafted ‘SVTH 2900’, self-grafted, grafted on ‘Multifort’; 2) Stems per plant: one, two, four. The experimental design was a randomized complete block with a 2x3 factorial arrangement with 3 replications (2020), and a 3x3 factorial arrangement with 4 replications (2021). Number of days and accumulated degree days from transplant as well as yield and average weight of fruits were recorded. Data were analyzed using an analysis of variance and Tukey's test. No differences were observed at the beginning of the harvest, while the productive cycle was prolonged by grafting and greater number of stems. Grafting increased the average weight of fruits, total yield by 32-34%, and the production of fruits weighing >150 g by 45%. Total yield was promoted in plants with two stems. In addition, the number of fruits was higher in plants with two stems in 2020, and increased in grafted plants with four stems in 2021. Under the trial conditions, the use of grafted tomato plants and plants with two stems is recommended since a better phenological and productive behavior is observed.

Abstract Until now, there is no protocol for the micropropagation of pecan trees (Carya illinoinensis) from explants of adult trees. Persistent microbial contamination and browning of tissues have impeded the success of previously proposed methods. Specifically, the adult tree leaf explant constitutes an unexplored starting point for the regeneration of C. illinoinensis. Therefore, the objective of this study was to determine a disinfection method and in vitro culture conditions that induce greater callus production in adult pecan tree leaf explants. Five disinfection methods were tested: (1) 70% ethanol for 50 s, 1.8-2.7% sodium hypochlorite with 2 drops/L of Tween 80 for 50 s, (2) 70% ethanol for 2 min, 1.8-2.7% sodium hypochlorite with 2 drops/L of Tween 80 for 2 min, (3) 70% ethanol for 2 min, 1.8-2.7% sodium hypochlorite with 2 drops/L of Tween 80 for 2 min, 1 g/L of carbendazim with 2 drops/L of Tween 80 for 20 min, (4) 70% ethanol for 2 min, 1.8-2.7% sodium hypochlorite with 2 drops/L of Tween 80 for 2 min, 8.8 g/L of carbendazim with 2 drops /L of Tween 80 for 20 min, and (5) 70% ethanol for 2 min, 1.8-2.7% sodium hypochlorite with 2 drops/L of Tween 80 for 2 min, 1 g/L of prochloraz with 2 drops/L of Tween 80 for 20 min. In addition, 6 different culture media were tested where the carbon source (sucrose, glucose and maltose), the inclusion of an antioxidant (AgNO3 and activated carbon), and the concentration of MS salts (maximum or one third of the maximum concentration) were varied. The explants were incubated for 50 days; a portion of the explants was incubated in the dark and the other under a photoperiod of 16 h. Likewise, some explants were inoculated in abaxial orientation, and others in abaxial orientation. The level of browning, the percentage of explants with callus, the fresh and dry weight of the calluses, and the percentage of explants contaminated by microorganisms were evaluated. The lowest percentage of contaminated explants was observed in the disinfection treatment that included immersion in prochloraz, and prochloraz was the only fungicide that allowed callogenesis in 100% of the treated explants. The darkening in the explants did not increase when the concentration of MS salts tripled. Glucose significantly reduced browning, but also significantly decreased the percentage of explants with callus, compared to that observed when using sucrose or maltose. The adaxial orientation promoted a higher percentage of explants with callus, with a fresh and dry weight, as well as a higher moisture percentage. The photoperiod prevented callus production, while incubation in the dark allowed more than 80% of the explants to produce callus in each treatment. The medium without antioxidant and the medium with 5 mg/L of silver nitrate gave similar browning results, while 1 g/L of activated carbon left the explants unviable. The present work proposes a method for the aseptic establishment and in vitro culture conditions for callus initiation in leaflets of adult pecan tree, with reproducible results.

Resumen Hasta ahora, no existe un protocolo para la micropropagación de nogal pecanero (Carya illinoinensis) a partir de explantes de árbol adulto. La contaminación microbiana persistente y el oscurecimiento oxidativo de los tejidos han impedido el éxito de métodos previamente propuestos. Específicamente, el explante de hoja de árbol adulto constituye un punto de partida inexplorado para la regeneración de C. illinoinensis. Por ello, el objetivo de este estudio fue determinar un método de desinfección y las condiciones de cultivo in vitro que inducen una mayor producción de callo en explantes de hoja de árbol adulto de nogal pecanero. Se probaron 5 métodos de desinfección: (1) etanol 70% por 50 s, 1.8-2.7% hipoclorito de sodio con 2 gotas/L de Tween 80 por 50 s, (2) etanol 70% por 2 min, 1.8-2.7% hipoclorito de sodio con 2 gotas/L de Tween 80 por 2 min, (3) etanol 70% por 2 min, 1.8-2.7% hipoclorito de sodio con 2 gotas/L de Tween 80 por 2 min, 1 g/L de carbendazim con 2 gotas/L de Tween 80 por 20 min, (4) etanol 70% por 2 min, 1.8-2.7% hipoclorito de sodio con 2 gotas/L de Tween 80 por 2 min, 8.8 g/L de carbendazim con 2 gotas/L de Tween 80 por 20 min, y (5) etanol 70% por 2 min, 1.8-2.7% hipoclorito de sodio con 2 gotas/L de Tween 80 por 2 min, 1 g/L de procloraz con 2 gotas/L de Tween 80 por 20 min. Además, se probaron 6 diferentes medios de cultivo donde se varió la fuente de carbono (sacarosa, glucosa y maltosa), la inclusión de un antioxidante (AgNO3 y carbón activado), y la concentración de sales MS (máxima o un tercio de la concentración máxima). Los explantes fueron incubados durante 50 días; una porción de los explantes fue incubada en la oscuridad y la otra ante un fotoperiodo de 16 h. Asimismo, unos explantes fueron inoculados en orientación abaxial, y otros en orientación abaxial. Se evaluó el oscurecimiento oxidativo de los explantes, el porcentaje de explantes con callo, el peso fresco y seco de los callos, y el porcentaje de explantes contaminados por microorganismos. El menor porcentaje de explantes contaminados se observó en el tratamiento de desinfección que incluyó inmersión en procloraz, además procloraz fue el único fungicida que permitió la callogénesis en el 100% de los explantes tratados. El oscurecimiento en los explantes no aumentó al triplicar la concentración de sales MS. La glucosa redujo significativamente el oscurecimiento de los explantes, pero también disminuyó de manera significativa el porcentaje de explantes con callo, en comparación con lo observado al utilizar sacarosa o maltosa. La orientación adaxial, promovió un mayor porcentaje de explantes con callo, con un peso fresco y seco, así como un porcentaje de humedad superiores. El fotoperiodo impidió la producción de callo, mientras que la incubación en la oscuridad permitió que más del 80% de los explantes produjeran callo. El medio sin antioxidante y el medio con 5 mg/L de nitrato de plata dieron resultados semejantes de oscurecimiento, mientras que 1 g/L de carbón activado dejó los explantes inviables. El presente trabajo propone un método para el establecimiento aséptico y condiciones de cultivo in vitro para la iniciación de callos en hojas de nogal pecanero adulto, con resultados reproducibles.

Abstract Yacon is mainly constituted of water and carbohydrates [single sugars and fructooligosaccharides (FOS)], thus being an excellent alternative for the growth and preservation of bacterial culture. Latilactobacillus sakei ACU-2 and Staphylococcus vitulinus ACU-10 comprised the autochthonous starter culture SAS-1 designed for the manufacture of dry sausages. This study evaluated the use of yacon juice as a potential growth medium and cryoprotectant for these bacteria. The growth medium was prepared with yacon juice supplemented with peptone and dipotassium phosphate. After growing, cells were resuspended in yacon juice (5, 10 and 25 mL/100 mL) and lyophilized. Viable cells were count before, immediately after lyophilization, and along 6 months of refrigerated storage. Both bacteria grew in every yacon concentration tested; however, juice concentration affected their growth. Latilactobacillus sakei grew at μ = 0.256 ± 0.01 giving the highest bacterial density at 10 mL/100 mL (Log DOmax 0.33 ± 0.01). While 5 mL/100 mL yacon juice provided the best conditions for S. vitulinus growth (μ = 0.215 ± 0.016; Log DOmax 0.32 ± 0.01). After lyophilization, the survival rate was 91.1% for L. sakei and 65.8% for S. vitulinus. Throughout storage, high cell counts suggested good stability of both bacteria. Results revealed that yacon juice comprises a nutritive substrate for the growth and cryopreservation of tested strains from the genus Latilactobacillus and Staphylococcus.

Resumo O yacon é constituído principalmente por água e carboidratos (açúcares simples e fruto-oligossacarídeos), sendo uma excelente alternativa para o crescimento e a preservação da cultura bacteriana. Latilactobacillus sakei ACU-2 e Staphylococcus vitulinus ACU-10 compuseram a cultura starter autóctone SAS-1, destinada à fabricação de embutidos secos. Este estudo avaliou o uso do suco de yacon como potencial meio de crescimento e crioprotetor para essas bactérias. O meio de crescimento foi preparado com suco de yacon suplementado com peptona e fosfato dipotássico. Após o crescimento, as células foram ressuspensas em suco de yacon (5, 10 e 25 mL/100 mL) e liofilizadas. As células viáveis foram contadas antes e imediatamente após a liofilização, e ao longo de seis meses de armazenamento refrigerado. Ambas as bactérias cresceram em cada concentração de yacon testada; no entanto, a concentração de suco afetou seu crescimento. Latilactobacillus sakei cresceu a μ = 0,256 ± 0,01, dando a maior densidade bacteriana em 10 mL/100 mL (Log DOmax 0,33 ± 0,01), enquanto a concentração 5 mL/100 mL de suco de yacon apresentou as melhores condições para o crescimento de S. vitulinus (μ = 0,215 ± 0,016; Log DOmax 0,32 ± 0,01). Após a liofilização, a taxa de sobrevivência foi de 91,1% para L. sakei e 65,8% para S. vitulinus. Ao longo do armazenamento, altas contagens de células sugeriram boa estabilidade de ambas as bactérias. Os resultados revelaram que o suco de yacon é um substrato nutritivo para o crescimento e a criopreservação das cepas testadas dos gêneros Latilactobacillus e Staphylococcus.

Abstract Spinal muscular atrophy (SMA) is considered one of the most common autosomal recessive disorders, with an estimated incidence of 1 in 10,000 live births. Testing for SMA has been recommended for inclusion in neonatal screening (NBS) panels since there are several therapies available and there is evidence of greater efficacy when introduced in the pre/early symptomatic phases. In Brazil, the National Neonatal Screening Program tests for six diseases, with a new law issued in 2021 stating that it should incorporate more diseases, including SMA. In the present study, dried blood spot (DBS) samples collected by the Reference Services of Neonatal Screening of RS and SP, to perform the conventional test were also screened for SMA, using real-time PCR, with SALSA MC002 technique. A total of 40,000 samples were analyzed, enabling the identification of four positive cases of SMA, that were confirmed by MLPA. Considering our sampling, Brazil seems to have an incidence comparable to the described in other regions. This work demonstrated that the use of the MC002 technique in samples routinely collected for the conventional NBS program is suitable to screen for SMA in our conditions and can be included in the expansion of the neonatal screening programs.