RITA selectively inhibits proliferation of BAP1-deficient cutaneous melanoma cells in vitro / 南方医科大学学报

OBJECTIVE@#To screen for small molecular compounds with selective inhibitory activity against cutaneous melanoma cells with BAP1 deletion.@*METHODS@#Cutaneous melanoma cells expressing wild-type BAP1 were selected to construct a BAP1 knockout cell model using CRISPR-Cas9 system, and small molecules with selective inhibitory activity against BAP1 knockout cells were screened from a compound library using MTT assay. Rescue experiment was carried out to determine whether the sensitivity of BAP1 knockout cells to the candidate compounds was directly related to BAP1 deletion. The effects of the candidate compounds on cell cycle and apoptosis were detected with flow cytometry, and the protein expressions in the cells were analyzed with Western blotting.@*RESULTS@#The p53 activator RITA from the compound library was shown to selectively inhibit the viability of BAP1 knockout cells. Overexpression of wild-type BAP1 reversed the sensitivity of BAP1 knockout cells to RITA, while overexpression of the mutant BAP1 (C91S) with inactivated ubiquitinase did not produce any rescue effect. Compared with the control cells expressing wild-type BAP1, BAP1 knockout cells were more sensitive to RITA-induced cell cycle arrest and apoptosis (P < 0.0001) and showed an increased expression of p53 protein, which was further increased by RITA treatment (P < 0.0001).@*CONCLUSION@#Loss of BAP1 results in the sensitivity of cutaneous melanoma cells to p53 activator RITA. In melanoma cells, the activity of ubiquitinase in BAP1 is directly related to their sensitivity to RITA. An increased expression of p53 protein induced by BAP1 knockout is probably a key reason for RITA sensitivity of melanoma cells, suggesting the potential of RITA as a targeted therapeutic agent for cutaneous melanoma carrying BAP1-inactivating mutations.

Seasonal analysis of food items and feeding habits of endangered riverine catfish Rita rita (Hamilton, 1822) / Análise sazonal de itens e hábitos alimentares de bagres ribeirinhos Rita rita (Hamilton, 1822) ameaçados de extinção

This investigation presents the food and feeding activity of and endangered riverine catfish Rita rita, during February 2017-January 2018. A total of 225 fish individuals was analyzed for stomach contents by characterizing the dominant food items and morphometric features. The results divulged ten major food items consumed, preferably fish scales and eggs, teleost fishes, copepods, cladocerans, rotifers, and mollusks. Total length and body weight of fish varied between 9-34 cm (20.53 ± 6.90 cm) and 10-400 g (9125.94 ± 102.07 g), respectively. The index of relative importance (IRI%) showed the importance of rotifers over the other food items. Morisita's index of diet overlap indicated seasonal variations in catfish diets with summer and monsoon displaying the least overlap, while maximum overlap during monsoon and winter seasons. The non-metric multidimensional scaling (nMDS) indicated the close association between the food items available during summer and winter seasons with a significant difference among the seasons (ANOSIM, R = 0.638, P = 0.013). Levin's niche breadth index arranged in the order of 0.88>0.81>0.78>0.63>0.43 for the size classes of V, IV, III, II and I, respectively. The PCA explained 95.39% of the total variance among the food items and fish size groups. Small-sized fish individuals displayed a greater correlation with food items suitable for their mouth size. In conclusion, the variety and frequency of food items recorded indicated considerable feeding plasticity and opportunistic feeding behavior with a shift from carnivorous to omnivorous feeding nature. This study could render useful information on the food and feeding habits of R. rita and provide background for preparing its diet for future aquaculture practices.

Esta investigação apresenta a alimentação e a atividade alimentar de bagres ribeirinhos Rita rita ameaçados de extinção, no período de fevereiro de 2017 a janeiro de 2018. Um total de 225 peixes foi analisado quanto ao conteúdo do estômago, caracterizando os itens alimentares dominantes e as características morfométricas. Os resultados apresentaram dez principais itens alimentares consumidos, preferencialmente escamas e ovos de peixes, teleósteos, copépodes, cladóceros, rotíferos e moluscos. O comprimento total e o peso corporal dos peixes variaram entre 9-34 cm (20,53 ± 6,90 cm) e 10-400 g (9125,94 ± 102,07 g), respectivamente. O índice de importância relativa (IRI%) mostrou a importância dos rotíferos sobre os demais itens alimentares. O índice de sobreposição de dieta de Morisita indicou variações sazonais nas dietas de bagres no verão e nas monções, exibindo a menor sobreposição, enquanto a sobreposição máxima ocorreu durante as estações das monções e do inverno. A escala multidimensional não métrica (nMDS) indicou a estreita associação entre os itens alimentares disponíveis durante o verão e o inverno, com uma diferença significativa entre as estações (ANOSIM, R = 0,638, P = 0,013). O índice de amplitude de nicho de Levin foi organizado na ordem de 0,88 > 0,81 > 0,78 > 0,63 > 0,43 para as classes de tamanho de V, IV, III, II e I, respectivamente. O PCA explicou 95,39% da variação total entre os itens alimentares e os grupos de tamanho de peixes. Peixes de pequeno porte apresentaram maior correlação com itens alimentares adequados ao tamanho da boca. Em conclusão, a variedade e a frequência dos itens alimentares registrados indicaram plasticidade alimentar considerável e comportamento alimentar oportunista, com uma mudança da natureza carnívora para onívora. Este estudo pode fornecer informações úteis sobre os hábitos alimentares e de alimentação de R. rita e apresentar subsídios para a preparação de sua dieta em futuras práticas de aquicultura.

Seasonal analysis of food items and feeding habits of endangered riverine catfish Rita rita (Hamilton, 1822) / Análise sazonal de itens e hábitos alimentares de bagres ribeirinhos Rita rita (Hamilton, 1822) ameaçados de extinção

This investigation presents the food and feeding activity of and endangered riverine catfish Rita rita, during February 2017-January 2018. A total of 225 fish individuals was analyzed for stomach contents by characterizing the dominant food items and morphometric features. The results divulged ten major food items consumed, preferably fish scales and eggs, teleost fishes, copepods, cladocerans, rotifers, and mollusks. Total length and body weight of fish varied between 9-34 cm (20.53 ± 6.90 cm) and 10-400 g (9125.94 ± 102.07 g), respectively. The index of relative importance (IRI%) showed the importance of rotifers over the other food items. Morisita's index of diet overlap indicated seasonal variations in catfish diets with summer and monsoon displaying the least overlap, while maximum overlap during monsoon and winter seasons. The non-metric multidimensional scaling (nMDS) indicated the close association between the food items available during summer and winter seasons with a significant difference among the seasons (ANOSIM, R = 0.638, P = 0.013). Levin’s niche breadth index arranged in the order of 0.88>0.81>0.78>0.63>0.43 for the size classes of V, IV, III, II and I, respectively. The PCA explained 95.39% of the total variance among the food items and fish size groups. Small-sized fish individuals displayed a greater correlation with food items suitable for their mouth size. In conclusion, the variety and frequency of food items recorded indicated considerable feeding plasticity and opportunistic feeding behavior with a shift from carnivorous to omnivorous feeding nature. This study could render useful information on the food and feeding habits of R. rita and provide background for preparing its diet for future aquaculture practices.

Esta investigação apresenta a alimentação e a atividade alimentar de bagres ribeirinhos Rita rita ameaçados de extinção, no período de fevereiro de 2017 a janeiro de 2018. Um total de 225 peixes foi analisado quanto ao conteúdo do estômago, caracterizando os itens alimentares dominantes e as características morfométricas. Os resultados apresentaram dez principais itens alimentares consumidos, preferencialmente escamas e ovos de peixes, teleósteos, copépodes, cladóceros, rotíferos e moluscos. O comprimento total e o peso corporal dos peixes variaram entre 9-34 cm (20,53 ± 6,90 cm) e 10-400 g (9125,94 ± 102,07 g), respectivamente. O índice de importância relativa (IRI%) mostrou a importância dos rotíferos sobre os demais itens alimentares. O índice de sobreposição de dieta de Morisita indicou variações sazonais nas dietas de bagres no verão e nas monções, exibindo a menor sobreposição, enquanto a sobreposição máxima ocorreu durante as estações das monções e do inverno. A escala multidimensional não métrica (nMDS) indicou a estreita associação entre os itens alimentares disponíveis durante o verão e o inverno, com uma diferença significativa entre as estações (ANOSIM, R = 0,638, P = 0,013). O índice de amplitude de nicho de Levin foi organizado na ordem de 0,88 > 0,81 > 0,78 > 0,63 > 0,43 para as classes de tamanho de V, IV, III, II e I, respectivamente. O PCA explicou 95,39% da variação total entre os itens alimentares e os grupos de tamanho de peixes. Peixes de pequeno porte apresentaram maior correlação com itens alimentares adequados ao tamanho da boca. Em conclusão, a variedade e a frequência dos itens alimentares registrados indicaram plasticidade alimentar considerável e comportamento alimentar oportunista, com uma mudança da natureza carnívora para onívora. Este estudo pode fornecer informações úteis sobre os hábitos alimentares e de alimentação de R. rita e apresentar subsídios para a preparação de sua dieta em futuras práticas de aquicultura.

Seasonal analysis of food items and feeding habits of endangered riverine catfish Rita rita (Hamilton, 1822)

Abstract This investigation presents the food and feeding activity of and endangered riverine catfish Rita rita, during February 2017-January 2018. A total of 225 fish individuals was analyzed for stomach contents by characterizing the dominant food items and morphometric features. The results divulged ten major food items consumed, preferably fish scales and eggs, teleost fishes, copepods, cladocerans, rotifers, and mollusks. Total length and body weight of fish varied between 9-34 cm (20.53 ± 6.90 cm) and 10-400 g (9125.94 ± 102.07 g), respectively. The index of relative importance (IRI%) showed the importance of rotifers over the other food items. Morisitas index of diet overlap indicated seasonal variations in catfish diets with summer and monsoon displaying the least overlap, while maximum overlap during monsoon and winter seasons. The non-metric multidimensional scaling (nMDS) indicated the close association between the food items available during summer and winter seasons with a significant difference among the seasons (ANOSIM, R = 0.638, P = 0.013). Levins niche breadth index arranged in the order of 0.88>0.81>0.78>0.63>0.43 for the size classes of V, IV, III, II and I, respectively. The PCA explained 95.39% of the total variance among the food items and fish size groups. Small-sized fish individuals displayed a greater correlation with food items suitable for their mouth size. In conclusion, the variety and frequency of food items recorded indicated considerable feeding plasticity and opportunistic feeding behavior with a shift from carnivorous to omnivorous feeding nature. This study could render useful information on the food and feeding habits of R. rita and provide background for preparing its diet for future aquaculture practices.

Resumo Esta investigação apresenta a alimentação e a atividade alimentar de bagres ribeirinhos Rita rita ameaçados de extinção, no período de fevereiro de 2017 a janeiro de 2018. Um total de 225 peixes foi analisado quanto ao conteúdo do estômago, caracterizando os itens alimentares dominantes e as características morfométricas. Os resultados apresentaram dez principais itens alimentares consumidos, preferencialmente escamas e ovos de peixes, teleósteos, copépodes, cladóceros, rotíferos e moluscos. O comprimento total e o peso corporal dos peixes variaram entre 9-34 cm (20,53 ± 6,90 cm) e 10-400 g (9125,94 ± 102,07 g), respectivamente. O índice de importância relativa (IRI%) mostrou a importância dos rotíferos sobre os demais itens alimentares. O índice de sobreposição de dieta de Morisita indicou variações sazonais nas dietas de bagres no verão e nas monções, exibindo a menor sobreposição, enquanto a sobreposição máxima ocorreu durante as estações das monções e do inverno. A escala multidimensional não métrica (nMDS) indicou a estreita associação entre os itens alimentares disponíveis durante o verão e o inverno, com uma diferença significativa entre as estações (ANOSIM, R = 0,638, P = 0,013). O índice de amplitude de nicho de Levin foi organizado na ordem de 0,88 > 0,81 > 0,78 > 0,63 > 0,43 para as classes de tamanho de V, IV, III, II e I, respectivamente. O PCA explicou 95,39% da variação total entre os itens alimentares e os grupos de tamanho de peixes. Peixes de pequeno porte apresentaram maior correlação com itens alimentares adequados ao tamanho da boca. Em conclusão, a variedade e a frequência dos itens alimentares registrados indicaram plasticidade alimentar considerável e comportamento alimentar oportunista, com uma mudança da natureza carnívora para onívora. Este estudo pode fornecer informações úteis sobre os hábitos alimentares e de alimentação de R. rita e apresentar subsídios para a preparação de sua dieta em futuras práticas de aquicultura.

Atuação fonoaudiológica no NASF do município de Santa Rita ­ PB / Speech-language therapy practice in NASF of the city of Santa Rita ­ PB / Atuación fonoaudiológica en la NASF del municipio de Santa Rita ­ PB

Introdução: O Núcleo de Apoio à Saúde da Família ­ NASF objetiva ampliar as ações de saúde na atenção básica e aumentar a resolubilidade de problemas clínico-sanitários nos territórios. Objetivo: Caracterizar a atuação fonoaudiológica no NASF do município de Santa Rita ­ PB. Método: Utilizou-se método descritivo e transversal, por intermédio de um questionário contendo 36 questões com respostas abertas e fechadas que investigavam o processo de trabalho no NASF. A coleta foi realizada com sete fonoaudiólogas, que atenderam aos seguintes critérios de elegibilidade: trabalhar no NASF há no mínimo seis meses e assinar o termo de consentimento livre e esclarecido. Os dados foram tabulados e analisados quantitativamente. Resultados: As fonoaudiólogas relataram realizar de maneira satisfatória, ações como PSE, Visita Domiciliar, Capacitação e Suporte aos ACS, Ações conjuntas com a ESF, Orientações, Atendimento individual e também evidenciaram a necessidade de contratação de mais fonoaudiólogos. Relataram ainda utilizar com frequência ferramentas como o Atendimento compartilhado com profissionais do NASF, o Apoio Matricial e a Clínica Ampliada. Sobre a infraestrutura, houve constância entre todas as profissionais percebendo que este quesito obteve uma pontuação pouco satisfatória. Conclusão: As fonoaudiólogas demonstraram sincronia no processo de trabalho e nas ações realizadas, declarando ainda que mesmo em meio à problemática encontrada no quesito infraestrutura, o NASF de Santa Rita ­ PB tem desempenhado seu papel de maneira efetiva.

Introduction: The Family Health Support Center - NASF aims to expand health care in basic care and increase the resolubility of clinical and health problems in the territories. Purpose: To characterize the speech-language therapy performance by the NASF in the city of Santa Rita ­ PB. Methods: A descriptive and cross-sectional method was used, through a questionnaire containing 36 questions with open and closed answers that investigated the work process in NASF. The collection was performed with seven speech therapists, who met the following eligibility criteria: working at NASF for at least six months and signing the informed consent form. Data were tabulated and analyzed quantitatively. Results: Some speech-language therapists have reported to realize in satisfactory way actions such as PSE, home visit, capacity and support to the ACS, join actions with the ESF, guidelines, individual care and they also pointed the need of speech-language therapists' employment. They also reported utilizing with frequency implements such as shared with NASF professionals, the matrix assistance and the amplified clinic. On the Infrastructure, there was constancy among all the professionals that this requirement occurred on a rather unsatisfactory way. Conclusion: The speech-language therapists have demonstrated synchrony in the process of working and in the actions taken, and they also declared that in spite of the precarious infrastructure situation, the NASF in Santa Rita ­ PB, has played its roll effectively.

Introducción: El Núcleo de Apoyo a la Salud de la Familia - NASF tiene como objetivo ampliar las acciones de salud en la atención básica y aumentar la resolución de problemas clínico-sanitarios en los territorios. Objetivo: Caracterizar la actuación fonoaudilógica em el NASF del município de Santa Rita ­ PB. Métodos: Se utilizó método descriptivo y transversal, a través de un cuestionario que contenía 36 preguntas con respuestas abiertas y cerradas que investigaban el proceso de trabajo en el NASF. La recolección fue realizada con siete fonoaudiólogas, que atendieron a los siguientes criterios de elegibilidad: trabajar en el NASF por lo menos seis meses y firmar el término de consentimiento libre y esclarecido. Los datos fueron tabulados y analizados cuantitativamente. Resultados: Las fonoaudiólogas relataron realizar de manera satisfactoria, acciones como PSE, Visita Domiciliaria, Capacitación y Apoyo a los ACS, Acciones conjuntas con la ESF, Directrices, Atención individual y también evidenciaron la necessidad de contratación de más fonoaudiólogos. Señalaron utilizar com frecuencia herramientas como el Servicio compartido com profesionales de la NASF, el Apoyo Matrical y la Clínica Ampliada. Sobre la infraestructua, hubo constância entre todas las profesionales al darse cuenta de que este punto há obtenido uma puntuación poco satisfactoria. Conclusión: Las fonoaudiólogas demostraron una sincronía en el proceso de trabajo y en las acciones realizadas, declarando que incluso en medio de la problemática encontrada en el ámbito de la infraestructura, el NASF de Santa Rita ­ PB ha desempeñado su papel de manera efectiva.

Medios de cultivo líquidos: un avance para la micropropagación comercial de zábila (Aloe barbadensis Mill.) / Liquid medium culture: an approach for the commercial micropropagation of aloe (Aloe barbadensis Mill.)

La micropropagación es una alternativa para la producción comercial de plantas de zábila (Aloe barbadensis Mill.) limitada por los altos costos de producción. Con el objetivo de prescindir de los agentes gelificantes, reduciendo costos, se comparó el medio de cultivo líquido con el medio de cultivo gelificado en las diferentes etapas de micropropagación de la zábila. En la etapa de establecimiento se observó mayor porcentaje de explantes contaminados en el medio de cultivo líquido estático (25.55 %) que en el medio gelificado (11.11 %); y aunque el resto de los explantes se establecieron independientemente de la condición del medio de cultivo, en el medio líquido alcanzaron mayor altura (3.81 cm) que en el medio gelificado (3.03 cm). En la etapa de multiplicación, la altura de los explantes (entre 4.43 y 6.01 cm) fue superior en los recipientes de inmersión temporal automatizado (RITA®) en comparación con el medio gelificado (entre 3.24 y 3.42 cm); sin diferencias significativas entre el número de brotes/explante. Todos los brotes enraizaron a los 30 días independientemente del medio de cultivo empleado (líquido estático y gelificado), sin observar variaciones en la altura del brote y, número y longitud de las raíces. El empleo de los medios de cultivo líquidos y la implementación de los sistemas de inmersión temporal tipo RITA® permiten reducir los costos de producción al prescindir de los agentes gelificantes, lo que representa un avance para la micropropagación comercial de zábila.

Micropropagation is considered a successful alternative for aloe (Aloe barbadensis Mill.) plant production. However, it has limited use due to the high production cost. Liquid media were compared to agar-gelled medium during all micropropagation stages of aloe to reduce the cost for gelling agent used. In the establishment stage, there was a higher percentage of contaminated explants in static liquid medium (25.55%) than those cultured in agar-gelled medium (11.11%), although all the explants were established independently of the culture medium used, higher height (3.81 cm) was observed in liquid medium than those growing in agar-gelled medium (3.03 cm). In the multiplication stage, explant height was higher in the recipients used for automated temporary immersion system (RITA®) (4.43 - 6.01 cm) than those cultured in agar-gelled medium (3.24 - 3.42 cm), there was no significant difference for number of shoots/explant. All shoots had roots at 30 days independently of used culture media (static liquid or agar-gelled media). Shoot height, number and root length had similar values in both culture media. The implementation of liquid media and automated temporary Immersion system RITA® may allow to reduce production costs of gelling agent used, it represents an approach for the commercial micropropagation of aloe.

Micropropagación de plantas de lechosa en recipientes de inmersión temporal a partir de brotes axilares / Micropropagation of papaya plants in temporary immersion recipients from axilary shoots

Se estandarizaron las condiciones de iniciación, multiplicación, enraizamiento y aclimatización de plantas hermafroditas de lechosa cv Maradol provenientes de brotes axilares, producidos en recipientes de inmersión temporal RITA®. En cada envase, contentivo de 200 ml de medio de cultivo líquido de Fitch, se colocaron cuatro brotes de 2 a 3 cm de longitud. Los biorreactores se conectaron a tres líneas de inmersión de 5, 2 y 1 min cada 4h y se colocaron 6 envases en promedio por línea, en condiciones de fotoperíodo de 16 h. Transcurridos 30 a 45 días, se cuantificaron los brotes y se clasificaron de acuerdo al tamaño: < 2 cm (pequeños), entre 2 a 3 cm (medianos), > 3 cm con y sin raíz (grandes). Los dos primeros tipos de brotes se continuaron multiplicando en los mismos medios; y los más elongados se aclimatizaron utilizando el Sistema Autotrófico Hidropónico (SAH). Se determinó la sanidad y la fidelidad de las plantas producidas mediante pruebas de ELISA y RAPD, respectivamente. Durante un periodo de 6 meses se reciclaron un total de 47 recipientes, los cuales produjeron 1.091 brotes: 377 pequeños; 482 medianos; 175 grandes sin raíz y 57 con raíz. Usando el SAH se obtuvo 89,5% de plantas aclimatizadas cuando se usaron brotes enraizados, y 41,6% a partir de brotes sin raíces. Con la combinación de las técnicas RITA y SAH se logró un sistema continuo y eficiente de producción de plantas sanas y fieles al tipo, en comparación con los métodos convencionales de micropropagación y aclimatización.

We standardized initiation, multiplication, rooting and acclimatization conditions of papaya cv Maradol hermaphrodite plants from axillary buds produced in temporary immersion reactor RITA®. Recipients contained 200 ml of Fitch liquid culture medium, and four shoots of 2 to 3 cm. in length were placed in each. The bioreactors were connected to three different immersion lines of 5, 2, and 1 min each 4h, with 6 containers per line on average, in 16 h photoperiod. After 30 to 45 days, the shoots produced were quantified and classified according to size: <2 cm (small), from 2 to 3 cm (medium), >3 cm with or without roots (large). The first two types of shoots were multiplied in the same culture media, and more elongated shoots were acclimatized using Autotrophic Hydroponic System (AHS). The sanity and fidelity of the produced plants were determined using ELISA and RAPD, respectively. For a period of six months 47 vessels were recycled and 1,091 shoots were produced: 377 small; 482 medium; 175 large without roots and 57 rooted shoots. Using AHS, 89.5% acclimatized plants were obtained when rooted shoots were used, and 41.6% from rootless shoots. With the combination of RITA and AHS techniques we achieved a continuous and efficient production of healthy and true to type papaya plants, in comparison to conventional micropropagation and acclimatization procedures.

Multiplicación in vitro de Psidium guajava L. en sistemas de inmersión temporal / In vitro multiplication of Psidium guajava L. in temporary immersion systems

La introducción de nuevos cultivares de guayabo (Psidium guajava L.) amerita su propagación masiva, lo cual solo puede ser satisfecho mediante la micropropagación. Sin embargo la micropropagación convencional dejó de ser económicamente eficiente, debido al uso de agentes gelificantes y el elevado número de operaciones manuales, por esta razón se planteó en esta investigación, generar una metodología que permita disminuir los costos de producción por la exclusión del gelificante en los medios de cultivo, evaluando los sistemas de inmersión temporal (SIT) en la multiplicación in vitro de guayabo. Para lo cual, se evaluó el efecto del cultivo en SIT, se comparó los SIT tipo BIT® y RITA® y se evaluó el tiempo (1 y 2 min) y frecuencia (3 y 4 veces/día) de inmersión. Luego de seis semanas de cultivo se evaluó: número de brotes (NB), numero de nudos (NN), longitud de brote (LB) y coeficiente de multiplicación (CM). Con el empleo de SIT se logró valores superiores para NB (2,17), NN (3,5), LB (10,7 mm) y CM (2,8). En la comparación entre SIT tipo RITA y BIT, valores superiores se obtuvieron con el RITA® para NB (3,8), NN (3,8), LB (16,6 mm) y CM (10,4). Se determinó que con 2 min de inmersión se logró los mayores valores de NB (3,7), NN (13,4), LB (15,3 mm) y con 2 min de inmersión 3-4 veces/día el mayor CM (9,4 y 10,4). Se concluye que el cultivo en RITA® en la multiplicación favoreció crecimiento y la proliferación de brotes de guayabo.

The introduction of new cultivars of guava (Psidium guajava L.) deserves its mass propagation, which can only be satisfied by micropropagation. However conventional micropropagation stopped being economically efficient due to the use of gelling agents and the high number of manual operations. For this reason was considered in this research, generate a methodology to reduce production costs by exclusion of gelling in culture media, assessing temporary immersion systems (TIS) in the in vitro multiplication of guava. For which, the effect of the culture way was evaluated in TIS, type TIB® and RITA® compared the TIS and was evaluated the time (1 and 2 min) and frequency (3 and 4 times / day) of immersions. After six weeks of culture were evaluated: shoots number (NS), nodes number (NN), shoot length (SL) and multiplication rate (MR). With the use of TIS higher values for NS (2.17), NN (3.5), SL (10.7 mm) and MC (2.8) was achieved. When comparing RITA® and TIB, higher values were obtained with the RITA® for NS (3.8), NN (3.8), SL (16.6 mm) and MC (10.4). It was determined that 2 min of immersion with the highest values of NS (3.7), NN (13.4), SL (15.3 mm) and 2 min immersion 3-4 times/day achieved the highest MC (9.4 and 10.4). We conclude that the RITA® culture favored the multiplication in growth and proliferation of shoots of guava.

Multiplicación en sistemas de inmersión temporal y enraizamiento ex vitro de ocumo blanco (Xanthosoma sagittifolium (L) Schott) / Multiplication in temporary immersion systems and ex vitro rooting of white cocoyam (Xanthosoma sagittifolium (L) Schott)

El ocumo (Xanthosoma sagittifollium (L.) Schott) es una Arácea cultivada en países tropicales debido al valor nutritivo de sus cormos. La principal limitante para su cultivo es la carencia de semilla de calidad, por esta razón se planteó evaluar la multiplicación de brotes de ocumo blanco en sistemas de inmersión temporal, y el enraizamiento ex vitro de los mismos, para lo cual se estudió el tiempo y la frecuencia de inmersión, y la densidad de explantes sobre la proliferación de los brotes. Asimismo, el efecto del ácido indolacético (AIA) y ácido indolbutírico (AIB) sobre el enraizamiento ex vitro de brotes. De acuerdo con los resultados obtenidos, la mayor eficiencia en la proliferación de brotes se obtuvo utilizando el sistema de inmersión temporal del tipo RITA®, con una frecuencia y tiempo de inmersión de 6 veces/día y 5 min, respectivamente, y una densidad de 9 explantes/RITA®. En el enraizamiento ex vitro se determinó que bajo las condiciones de cultivo empleadas no es necesario el uso de auxinas. Se concluye que es posible la multiplicación eficiente de ocumo blanco en sistemas de inmersión temporal, y realizar el enraizamiento ex vitro sin el uso de auxinas.

The white cocoyam (Xanthosoma sagittifollium (L.) Schott), is an Arácea cultivated in tropical countries, due to the nutritional value of its corms. The main limiting factor for cultivation is the lack of healthy seed, by this reason be outlined to evaluate the multiplication of shoots of white cocoyam in temporary immersion systems and the ex vitro rooting of the same. For that which, itself study, the time and frequency of immersion and the density of explants on the proliferation of the shoots. As well as, the effect of the indole acetic acid (IAA) and indole butyric acid (IBA) on ex vitro rooting the shoots was studied. According to the results obtained, the greater efficiency in the proliferation of shoots was obtained utilizing the temporary immersion system of the type RITA®, with a frequency and time of immersion of 6 times/day and 5 min, respectively and a density of 9 explantes/RITA®. In the ex vitro rooting was determined that under the conditions of employed cultivation is not necessary the use of auxins. It is concluded that is possible the efficient multiplication of white cocoyam in temporary immersion systems and to carry out the ex vitro rooting without the use of auxins.

Lens proteome map and -crystallin profile of the catfish Rita rita.

Crystallins are a diverse group of proteins that constitute nearly 90% of the total soluble proteins of the vertebrate eye lens and these tightly packed crystallins are responsible for transparency of the lens. These proteins have been studied in different model and non-model species for understanding the modifications they undergo with ageing that lead to cataract, a disease of protein aggregation. In the present investigation, we studied the lens crystallin profile of the tropical freshwater catfish Rita rita. Profiles of lens crystallins were analyzed and crystallin proteome maps of Rita rita were generated for the first time. A-crystallins, member of the -crystallin family, which are molecular chaperons and play crucial role in maintaining lens transparency were identified by 1-and 2-D immunoblot analysis with anti-A-crystallin antibody. Two protein bands of 19-20 kDa were identified as A-crystallins on 1-D immunoblots and these bands separated into 10 discrete spots on 2-D immunoblot. However, anti-B-crystallin and antiphospho-B-crystallin antibodies were not able to detect any immunoreactive bands on 1- and 2-D immunoblots, indicating B-crystallin was either absent or present in extremely low concentration in Rita rita lens. Thus, Rita rita -crystallins are more like that of the catfish Clarias batrachus and the mammal kangaroo in its A- and B-crystallin content (contain low amount from 5-9% of aB-crystallin) and unlike the dogfish, zebrafish, human, bovine and mouse -crystallins (contain higher amount of B-crystallin from 25% in mouse and bovine to 85% in dogfish). Results of the present study can be the baseline information for stimulating further investigation on Rita rita lens crystallins for comparative lens proteomics. Comparing and contrasting the -crystallins of the dogfish and Rita rita may provide valuable information on the functional attributes of A- and B-isoforms, as they are at the two extremes in terms of A-and B-crystallin content.

Fitossociologia de um Cerrado denso em área de influência de torre de fluxo de carbono, Pé-de-Gigante, Parque Estadual de Vassununga, SP / Phytossociology of a dense Cerrado on the footprint of a carbon flux tower, Pé-de-Gigante, Vassununga State Park, SP

O domínio do Cerrado compreende uma área contínua nos estados centrais do Brasil e áreas disjuntas em outros estados, incluindo São Paulo. Essa vegetação ocupava originalmente 21 por cento do território brasileiro, restando atualmente apenas 21,6 por cento de sua extensão original. A área recoberta por essa vegetação em São Paulo cobria 14 por cento de sua área total e seus remanescentes recobrem menos de 1 por cento da ocorrência original dessa vegetação. Estudos recentes indicam que o valor da produtividade líquida no Cerrado Pé-de-Gigante (SP) constitui um pequeno dreno de carbono e indicou que a sazonalidade foi o fator determinante do valor observado. Os estudos dos fluxos de carbono em ecossistemas terrestres são raramente acompanhados de abordagens ecofisiológicas de modo a explorar a relação funcional das espécies que compõem o ecossistema e os valores líquidos obtidos para o mesmo. Assim, o objetivo deste trabalho foi caracterizar estruturalmente a vegetação presente na área de maior influência da torre de fluxo instalada no Cerrado Pé-de-Gigante, visando possibilitar estudos relacionados à quantificação em longo prazo da dinâmica dos fluxos de água, energia e CO2 na vegetação de Cerrado. Para isso foram levantadas 20 parcelas (10 x 10 m) em 0,2 ha de Cerrado, e amostraram-se todas as plantas com perímetro ao nível do solo >6 cm (exceto lianas e árvores mortas). A distribuição das classes de diâmetro e estrutura vertical, assim como os parâmetros fitossociológicos foram analisados. Encontramos 1451 indivíduos, distribuídos em 85 espécies, 52 gêneros e 31 famílias. A densidade absoluta e área basal foram de 7255 ind. ha-1 e de 7,9 m².ha-1, respectivamente. A família Leguminosae apresentou o maior número de espécies (13). O Índice de diversidade de Shannon (H') foi 3,27 nats.ind-1. A distribuição em classes de diâmetro mostrou uma curva de "J" invertido, estando a maioria dos indivíduos na primeira classe. Concluímos que a área deve ser classificada como Cerrado denso, devido principalmente à dominância pela espécie arbórea Anadenanthera falcata, cuja ocorrência no estado foi relatada apenas em locais com solos ricos em saturação de bases na região das Cuestas Basálticas, devido também à maior área basal dos indivíduos, comparando com outros fragmentos de Cerrado. Além da espécie citada, Myrcia lingua e Xylopia aromatica, apresentaram os maiores IVI (Valor de importância).

The Cerrado vegetation covers the Brazilian central areas (core area) which include extensive and continuous areas in the central States and some marginal areas in other States, including São Paulo, accounting originally for 21 percent of Brazilian territory. Nowadays, the remnants are 21,6 percent of its original coverage. About of 14 percent of the São Paulo State territory was originally covered with Cerrado vegetation, and today remnants account for less than 1 percent of the original distribution. Recent studies indicate that the net productivity in the Pé-de-Gigante Cerrado constitutes a small carbon sink, with seasonality playing a major role. The carbon flux studies in terrestrial ecosystems have rarely been accompanied by ecophysiological studies in order to explore the functional relationships among species, ecosystem and the values achieved. Therefore, our objectives were to characterize the vegetation structure in the meteorological and flux tower footprint. The flux tower allows studies related to long term measurements and dynamics of water, solar energy and CO2 fluxes in Cerrado vegetation. Plots (10 x 10 m) were established in 0.2 ha area of Cerrado and all vascular plants with stem perimeter at ground level >6 cm (except vines and dead individuals) were sampled. The distribution of diameter classes and vertical structure, as well as the phytosociological parameters were analyzed. We recorded 1,451 individuals, distributed in 85 species belonging to 31 different families. The absolute density and the basal area were 7,255 ind.ha-1 and 7.9 m².ha-1, respectively. Leguminosae showed the highest number of species (13). The Shannon's Index was 3.27. The distribution of diameter classes showed a curve in inverted "J", most of the individuals being in the initial classes. We concluded that the area must be classified as dense Cerrado, mainly due to Anadenanthera falcata dominance, which occurrence in São Paulo State was reported only in rich soil sites in Cuestas Basaltic regions and also due to major basal area of these individuals, comparing with other Cerrado vegetation sites. A. falcata, Myrcia lingua and Xylopia aromatica showed the highest IVI (Importance value index).

Preponderance of GC-rich sites in silver-stained nucleolus organizing regions of Rita rita (Hamilton) and Mystus gulio (Hamilton) (Bagridae, Pisces), as revealed by chromomycin A3-staining technique and scanning electron microscopic studies

The karyotypes of two species of catfish, Rita rita (Hamilton) (2n = 54; 14m + 34sm + 6st; NF = 102) and Mystus gulio (Hamilton) (2n = 58; 30m + 12sm + 2st + 14t, NF = 100) were studied through Giemsa-, silver- and chromomycin A(3)-staining techniques. The silver-stained karyotypes in both sexes of R. rita and M. gulio revealed that the nucleolus organizing regions were located terminally at the shorter arms (Tp) of one pair of submetacentric chromosomes, placed at positions Nos. 2 and 1, respectively, which was confirmed by scanning electron microscopy. Staining with a GC-specific fluorochrome, chromomycin A(3), produced bright fluorescence in the Ag-positive nucleolus organizer regions, suggesting thereby that nucleolus organizing regions actually included GC-rich sites of active r-RNA genes in metaphase chromosomes of these two bagrids. Further such studies are needed due to the extreme paucity of data on fish.

Propagación in vitro de Phalaenopsis (Orchidaceae) a partir de protocormos, mediante el sistema de inmersión temporal RITA / In vitro propagation of Phalaenopsis (Orchidaceae) from protocorms, using the temporary inmersion system RITA

Protocormos in vitro de Phalaenopsis de tres meses de edad se transfirieron a contenedores RITA® con el fin depropagarlos masivamente. Los factores evaluados fueron la concentración de sacarosa en el medio y la frecuencia de inmersión. Se dispusieron cinco pares de contenedores RITA® con medio de cultivo líquido a concentraciones de sacarosa de 0, 15, 30, 45 y 60 g/L. El medio utilizado fue el MS a la mitad de la concentración de las sales, suplementado con vitaminas y tidiazuron (5 mg/L). El experimento se realizó en dos etapas, cada una con duración de dos meses. La primera etapa con una frecuencia de inmersión de cuatro horas y la segunda con una frecuencia de inmersión de ocho horas, ambas con un tiempo de inmersión de un minuto. Los resultados mostraron que la mejor respuesta proliferativa, con 8,2 protocormos adventicios por protocormo por mes, se obtuvo en el medio con 15 g/L de sacarosa y un tiempo de inmersión de un minutocada cuatro horas.

In order to massively propagate Phalaenopsis orchids, three months old in vitro protocorms were transferred to RITA® vessels. The evaluation factors were the sucrose concentration in the culture medium and the frequency immersion. There was a set of five pairs of RITA® vessels with liquid culture medium containing sucrose at 0, 15, 30, 45, 60 g/L. A half-strength MS medium plus vitamins, supplemented with vitamins and thidiazuron (5 mg/L) was used. The experiment had two stages, each lasting two months. Each stage had a one minute immersion. The first stage had a four hour immersion frequency and the second one had an eight hour immersion frequency. Results showed that the best proliferating response was reached in a 15 g/L of sucrose medium with one minute of immersion time every four hours, resulting in 8,2 adventitious protocorms per protocorm per month.

Multimodality Treatment of Malignant Nonfunctioning Islet Cell Tumor of the Pancreas: A Case Report / 한국간담췌외과학회지

Nonfunctioning islet cell tumor of the pancreas is mostly malignant and has a poor prognosis. But, even in case of hepatic metastasis, the multimodality treatment including surgical resection, transhepatic arterial chemoembolization(TACE), radiofrequency interstitial tissue ablation(RITA), and systemic chemotherapy provides a better prognosis. We report a case of the multimodality treatment for malignant nonfunctioning islet cell tumor of the pancreas. The patient was 38-year old male whose diagnosis was made by routine health check-up. Abdominal computed tomography revealed about 20cm sized hypervascular mass in the tail of the pancreas. After preoperative selective arterial embolization for the purpose of reducing surgical bleeding, we explored his abdomen and found the mass originating from the tail of the pancreas and expanding to the spleen and transverse colon. We performed distal pancreatectomy combined with total gastrectomy, splenectomy, and transverse colectomy. Also, we found multiple bilobar hepatic metastatic nodules and performed the wedge resection for the lesion of the left lateral segment. Postoperatively, we underwent the TACE and the RITA for the remaining right hepatic metastatic nodules, then performed the systemic chemotherapy with interferon-alpha. At postoperative 22 months, the patient was alive with regular follow up of viable right hepatic metastatic nodules.