Automating behavioral analysis in neuroscience: Development of an open-source python software for more consistent and reliable results.

BACKGROUND: The application of automated analyses in neuroscience has become a practical approach. With automation, the algorithms and tools employed perform fast and accurate data analysis. It minimizes the inherent errors of manual analysis performed by a human experimenter. It also reduces the time required to analyze a large amount of data and the need for human and financial resources. METHODS: In this work, we describe a protocol for the automated analysis of the Morris Water Maze (MWM) and the Open Field (OF) test using the OpenCV library in Python. This simple protocol tracks mice navigation with high accuracy. RESULTS: In the MWM, both automated and manual analysis revealed similar results regarding the time the mice stayed in the target quadrant (p = 0.109). In the OF test, both automated and manual analysis revealed similar results regarding the time the mice stayed in the center (p = 0.520) and in the border (p = 0.503) of the field. CONCLUSIONS: The automated analysis protocol has several advantages over manual analysis. It saves time, reduces human errors, can be customized, and provides more consistent information about animal behavior during tests. We conclude that the automated protocol described here is reliable and provides consistent behavioral analysis in mice. This automated protocol could lead to deeper insight into behavioral neuroscience.

Ethnobotanical and ethnopharmacological study of medicinal plants used by a traditional community in Brazil's northeastern / Estudo etnobotânico e etnofarmacológico de plantas medicinais utilizadas por uma comunidade tradicional do nordeste brasileiro

The aim of this research is to make a survey of the socio-environmental characteristics and the ethnobotanical study of medicinal plants used in a traditional community in the Brazilian Northeast, Alagoas. The study was made based on visits with the application of a questionnaire with questions related to the socio-economic element and on the diversity of plants used in herbal medicine. The research was made from March/2019 to February/2020, where families and interviewed plant exhibitors were interviewed for botanical identification. The studied community, which were 24 interviewees, was compiled by residents of the Quilombola community from Pau D'arco in Arapiraca city - Alagoas. Residents interviewed, 15 (62.5%) attended between 56 to 80 years, 11 interviewees about 46% were born in the community and 13 (54%) had a fundamentally incomplete nature. At the end, there were mentioned 30 plant species used for phytotherapeutic purposes, from which presents bigger usage as plants against arterial hypertension (Salvia rosmarinus Schleid), diabetes Mellitos (Croton heliotropiifolius Kunth), pain and inflammation (Alternanthera tenella Colla), present the biggest number of species in the community. The species cited are related to numerous medicinal uses, among which there will be predominant associations associated with cardiovascular and inflammatory processes. The tea is the main way of preparing plants. It is perceived that medicinal plants are only widely used by this Quilombola community of and growth of the crops in the backyard are considered a tradition.

Esta pesquisa teve como objetivo realizar um levantamento das características socioambientais e estudo etnobotânico de plantas medicinais utilizadas por uma comunidade tradicional do nordeste brasileiro, Alagoas, Brasil. O estudo se deu por meio de visitas com aplicação de um questionário contendo perguntas relacionadas aos fatores socio-econômicos e sobre a diversidade das plantas utilizadas como medicinais. O trabalho foi realizado no período de março/2019 a fevereiro/2020, onde foram entrevistadas famílias e catalogadas amostras de plantas para identificação botânica. A população estudada, com 24 entrevistados, foi composta por moradores da comunidade Quilombola Pau D'arco da cidade de Arapiraca - Alagoas. Dos moradores entrevistados, 15 (62,5%) apresentavam idade entre 56 e 80 anos, 11 dos entrevistados cerca de 46%, são naturais da comunidade e 13 (54%) possuíam ensino fundamental incompleto. Ao todo, foram citadas 30 espécies de plantas utilizadas para fins terapêuticos, das quais apresentam maior utilização as plantas contra a hipertensão arterial (Salvia rosmarinus Schleid), diabetes Mellitos (Croton heliotropiifolius Kunth), dor e inflamação (Alternanthera tenella Colla). As espécies citadas estão relacionadas a inúmeras utilizações medicinais, entre os quais predominaram as doenças associadas ao aparelho cardiovascular e processos inflamatórios. O chá é principal forma de preparo das plantas. Percebe-se que as plantas medicinais são amplamente utilizadas por essa comunidade quilombola e o cultivo no quintal é considerado uma tradição.

Ethnobotanical and ethnopharmacological study of medicinal plants used by a traditional community in Brazil's northeastern / Estudo etnobotânico e etnofarmacológico de plantas medicinais utilizadas por uma comunidade tradicional do nordeste brasileiro

The aim of this research is to make a survey of the socio-environmental characteristics and the ethnobotanical study of medicinal plants used in a traditional community in the Brazilian Northeast, Alagoas. The study was made based on visits with the application of a questionnaire with questions related to the socio-economic element and on the diversity of plants used in herbal medicine. The research was made from March/2019 to February/2020, where families and interviewed plant exhibitors were interviewed for botanical identification. The studied community, which were 24 interviewees, was compiled by residents of the Quilombola community from Pau D'arco in Arapiraca city - Alagoas. Residents interviewed, 15 (62.5%) attended between 56 to 80 years, 11 interviewees about 46% were born in the community and 13 (54%) had a fundamentally incomplete nature. At the end, there were mentioned 30 plant species used for phytotherapeutic purposes, from which presents bigger usage as plants against arterial hypertension (Salvia rosmarinus Schleid), diabetes Mellitos (Croton heliotropiifolius Kunth), pain and inflammation (Alternanthera tenella Colla), present the biggest number of species in the community. The species cited are related to numerous medicinal uses, among which there will be predominant associations associated with cardiovascular and inflammatory processes. The tea is the main way of preparing plants. It is perceived that medicinal plants are only widely used by this Quilombola community of and growth of the crops in the backyard are considered a tradition.

Esta pesquisa teve como objetivo realizar um levantamento das características socioambientais e estudo etnobotânico de plantas medicinais utilizadas por uma comunidade tradicional do nordeste brasileiro, Alagoas, Brasil. O estudo se deu por meio de visitas com aplicação de um questionário contendo perguntas relacionadas aos fatores socio-econômicos e sobre a diversidade das plantas utilizadas como medicinais. O trabalho foi realizado no período de março/2019 a fevereiro/2020, onde foram entrevistadas famílias e catalogadas amostras de plantas para identificação botânica. A população estudada, com 24 entrevistados, foi composta por moradores da comunidade Quilombola Pau D’arco da cidade de Arapiraca - Alagoas. Dos moradores entrevistados, 15 (62,5%) apresentavam idade entre 56 e 80 anos, 11 dos entrevistados cerca de 46%, são naturais da comunidade e 13 (54%) possuíam ensino fundamental incompleto. Ao todo, foram citadas 30 espécies de plantas utilizadas para fins terapêuticos, das quais apresentam maior utilização as plantas contra a hipertensão arterial (Salviaros marinus Schleid), diabetes Mellitos (Croton heliotropiifolius Kunth), dor e inflamação (Alternanthera tenella Colla). As espécies citadas estão relacionadas a inúmeras utilizações medicinais, entre os quais predominaram as doenças associadas ao aparelho cardiovascular e processos inflamatórios. O chá é principal forma de preparo das plantas. Percebe-se que as plantas medicinais são amplamente utilizadas por essa comunidade quilombola e o cultivo no quintal é considerado uma tradição.

Ethnobotanical and ethnopharmacological study of medicinal plants used by a traditional community in Brazils northeastern

Abstract The aim of this research is to make a survey of the socio-environmental characteristics and the ethnobotanical study of medicinal plants used in a traditional community in the Brazilian Northeast, Alagoas. The study was made based on visits with the application of a questionnaire with questions related to the socio-economic element and on the diversity of plants used in herbal medicine. The research was made from March/2019 to February/2020, where families and interviewed plant exhibitors were interviewed for botanical identification. The studied community, which were 24 interviewees, was compiled by residents of the Quilombola community from Pau Darco in Arapiraca city - Alagoas. Residents interviewed, 15 (62.5%) attended between 56 to 80 years, 11 interviewees about 46% were born in the community and 13 (54%) had a fundamentally incomplete nature. At the end, there were mentioned 30 plant species used for phytotherapeutic purposes, from which presents bigger usage as plants against arterial hypertension (Salvia rosmarinus Schleid), diabetes Mellitos (Croton heliotropiifolius Kunth), pain and inflammation (Alternanthera tenella Colla), present the biggest number of species in the community. The species cited are related to numerous medicinal uses, among which there will be predominant associations associated with cardiovascular and inflammatory processes. The tea is the main way of preparing plants. It is perceived that medicinal plants are only widely used by this Quilombola community of and growth of the crops in the backyard are considered a tradition.

Resumo Esta pesquisa teve como objetivo realizar um levantamento das características socioambientais e estudo etnobotânico de plantas medicinais utilizadas por uma comunidade tradicional do nordeste brasileiro, Alagoas, Brasil. O estudo se deu por meio de visitas com aplicação de um questionário contendo perguntas relacionadas aos fatores socio-econômicos e sobre a diversidade das plantas utilizadas como medicinais. O trabalho foi realizado no período de março/2019 a fevereiro/2020, onde foram entrevistadas famílias e catalogadas amostras de plantas para identificação botânica. A população estudada, com 24 entrevistados, foi composta por moradores da comunidade Quilombola Pau Darco da cidade de Arapiraca - Alagoas. Dos moradores entrevistados, 15 (62,5%) apresentavam idade entre 56 e 80 anos, 11 dos entrevistados cerca de 46%, são naturais da comunidade e 13 (54%) possuíam ensino fundamental incompleto. Ao todo, foram citadas 30 espécies de plantas utilizadas para fins terapêuticos, das quais apresentam maior utilização as plantas contra a hipertensão arterial (Salvia rosmarinus Schleid), diabetes Mellitos (Croton heliotropiifolius Kunth), dor e inflamação (Alternanthera tenella Colla). As espécies citadas estão relacionadas a inúmeras utilizações medicinais, entre os quais predominaram as doenças associadas ao aparelho cardiovascular e processos inflamatórios. O chá é principal forma de preparo das plantas. Percebe-se que as plantas medicinais são amplamente utilizadas por essa comunidade quilombola e o cultivo no quintal é considerado uma tradição.

Ethnobotanical and ethnopharmacological study of medicinal plants used by a traditional community in Brazil's northeastern.

The aim of this research is to make a survey of the socio-environmental characteristics and the ethnobotanical study of medicinal plants used in a traditional community in the Brazilian Northeast, Alagoas. The study was made based on visits with the application of a questionnaire with questions related to the socio-economic element and on the diversity of plants used in herbal medicine. The research was made from March/2019 to February/2020, where families and interviewed plant exhibitors were interviewed for botanical identification. The studied community, which were 24 interviewees, was compiled by residents of the Quilombola community from Pau D'arco in Arapiraca city - Alagoas. Residents interviewed, 15 (62.5%) attended between 56 to 80 years, 11 interviewees about 46% were born in the community and 13 (54%) had a fundamentally incomplete nature. At the end, there were mentioned 30 plant species used for phytotherapeutic purposes, from which presents bigger usage as plants against arterial hypertension (Salvia rosmarinus Schleid), diabetes Mellitos (Croton heliotropiifolius Kunth), pain and inflammation (Alternanthera tenella Colla), present the biggest number of species in the community. The species cited are related to numerous medicinal uses, among which there will be predominant associations associated with cardiovascular and inflammatory processes. The tea is the main way of preparing plants. It is perceived that medicinal plants are only widely used by this Quilombola community of and growth of the crops in the backyard are considered a tradition.

Genetic Diversity in Nannotrigona testaceicornis (Hymenoptera: Apidae) Aggregations in Southeastern Brazil.

The Meliponini, also known as stingless bees, are distributed in tropical and subtropical areas of the world and plays an essential role in pollinating many wild plants and crops These bees can build nests in cavities of trees or walls, underground or in associations with ants or termites; interestingly, these nests are sometimes found in aggregations. In order to assess the genetic diversity and structure in aggregates of Nannotrigona testaceicornis (Lepeletier), samples of this species were collected from six aggregations and genetically analyzed for eight specific microsatellite loci. We observed in this analysis that the mean genetic diversity value among aggregations was 0.354, and the mean expected and observed heterozygosity values was 0.414 and 0.283, respectively. The statistically significant Fis value indicated an observed heterozygosity lower than the expected heterozygosity in all loci studied resulting in high homozygosis level in these populations. In addition, the low number of private alleles observed reinforces the absence of structuring that is seen in the aggregates. These results can provide relevant information about genetic diversity in aggregations of N. testaceicornis and contribute to the management and conservation of these bees' species that are critical for the pollination process.

Analysis of food habits of skate Rioraja agassizii (Elasmobranchii, Rajidae) from southern Brazil / Análise quantitativa dos hábitos alimentares da raia Rioraja agassizii (Elasmobranchii, Rajidae) do Sudeste e Sul do Brasil

Abstract Catches and exports of skate Rioraja agassizii place this species as “vulnerable to extinction” on the IUCN Red List; therefore, biological and ecological knowledge becomes an important instrument for its conservation control. This study described and quantified the diet composition of R. agassizii by means of stomach analysis contents in the periods 2005-2006 and 2012-2013. We analyzed and quantified stomach contents in terms of abundance (%N), weight (%M), frequency of occurrence (% FO), and index of relative importance (IRI). The results showed differences in the food rates between the periods. However, the groups of food items were the same: Teleostei fish, decapods, and mollusks. In 2005-2006, the diet consisted mainly of shrimp, however, in 2012-2013 it consisted of fish, followed by decapods, especially shrimps. The differences in diets may be attributed to shrimp abundance, which do not characterize a change in the eating habits in 2012-2013, because, in addition to fish, shrimps were also important food sources. The presence of a certain prey is more related to its availability rather than the feeding preference of skate. The amount of ingested items is associated to biological and environmental factors, so that further studies relating diet with capture area, seasonality, depth, and other factors should be conducted.

Resumo A raia-santa, Rioraja agassizii é uma espécie endêmica da qual pouco se conhece sobre sua biologia e ecologia, o que a leva a ser classificada como “vulnerável a extinção”. O objetivo foi identificar e quantificar a composição da dieta alimentar da R. agassizii através da análise do conteúdo estomacal de exemplares capturados nos períodos de 2005-2006 e 2012-2013. Foram analisados os conteúdos alimentares e quantificados em abundância (N%), peso (M%), frequência de ocorrência (FO%) e índice de importância relativa (IRI) de cada item. Os resultados mostraram que em 2012-13 a dieta foi composta por peixes, seguido de decápodes, especialmente camarões, entretanto em 2005-06 foi composta basicamente por camarões. A diferença nas dietas não quer dizer que os hábitos alimentares mudaram, pois em 2012-13 além dos peixes, os camarões também foram classificados como maior importância alimentar. Considerando que a quantidade e variedade dos itens ingeridos está relacionada, principalmente, a fatores ambientais são necessários mais estudos que relacionem áreas de captura, sazonalidade e profundidade.

Analysis of food habits of skate Rioraja agassizii (Elasmobranchii, Rajidae) from southern Brazil.

Catches and exports of skate Rioraja agassizii place this species as "vulnerable to extinction" on the IUCN Red List; therefore, biological and ecological knowledge becomes an important instrument for its conservation control. This study described and quantified the diet composition of R. agassizii by means of stomach analysis contents in the periods 2005-2006 and 2012-2013. We analyzed and quantified stomach contents in terms of abundance (%N), weight (%M), frequency of occurrence (% FO), and index of relative importance (IRI). The results showed differences in the food rates between the periods. However, the groups of food items were the same: Teleostei fish, decapods, and mollusks. In 2005-2006, the diet consisted mainly of shrimp, however, in 2012-2013 it consisted of fish, followed by decapods, especially shrimps. The differences in diets may be attributed to shrimp abundance, which do not characterize a change in the eating habits in 2012-2013, because, in addition to fish, shrimps were also important food sources. The presence of a certain prey is more related to its availability rather than the feeding preference of skate. The amount of ingested items is associated to biological and environmental factors, so that further studies relating diet with capture area, seasonality, depth, and other factors should be conducted.

Glycol methacrylate embedding for the histochemical study of the gastrointestinal tract of dogs naturally infected with Leishmania infantum.

In canine visceral leishmaniasis a diffuse chronic inflammatory exudate and an intense parasite load throughout the gastrointestinal tract has been previously reported. However, these studies did not allow a properly description of canine cellular morphology details. The aim of our study was to better characterize these cells in carrying out a qualitative and quantitative histological study in the gastrointestinal tract of dogs naturally infected with Leishmania infantum by examining gut tissues embedded in glycol methacrylate. Twelve infected adult dogs were classified in asymptomatic and symptomatic. Five uninfected dogs were used as controls. After necropsy, three samples of each gut segment, including esophagus, stomach, duodenum, jejunum, ileum, cecum, colon, and rectum were collected and fixed in Carnoy's solution for glycol methacrylate protocols. Sections were stained with hematoxylin-eosin, toluidine blue borate, and periodic acid-Schiff stain. Leishmania amastigotes were detected by immunohistochemistry employed in both glycol methacrylate and paraffin embedded tissues. The quantitative histological analysis showed higher numbers of plasma cells, lymphocytes and macrophages in lamina propria of all segments of GIT of infected dogs than controls. The parasite load was more intense and cecum and colon, independently of the clinical status of these dogs. Importantly, glycol methacrylate embedded tissue stained with toluidine blue borate clearly revealed mast cell morphology, even after mast cell degranulation. Infected dogs showed lower numbers of mast cells in all gut segments than did controls. Despite the glycol methacrylate (GMA) protocol requires more attention and care than the conventional paraffin processing, this embedding procedure proved to be especially suitable for the present histological study, where it allowed to preserve and observe cell morphology in fine detail.

Myosin Va is developmentally regulated and expressed in the human cerebellum from birth to old age.

Myosin Va functions as a processive, actin-based motor molecule highly enriched in the nervous system, which transports and/or tethers organelles, vesicles, and mRNA and protein translation machinery. Mutation of myosin Va leads to Griscelli disease that is associated with severe neurological deficits and a short life span. Despite playing a critical role in development, the expression of myosin Va in the central nervous system throughout the human life span has not been reported. To address this issue, the cerebellar expression of myosin Va from newborns to elderly humans was studied by immunohistochemistry using an affinity-purified anti-myosin Va antibody. Myosin Va was expressed at all ages from the 10th postnatal day to the 98 th year of life, in molecular, Purkinje and granular cerebellar layers. Cerebellar myosin Va expression did not differ essentially in localization or intensity from childhood to old age, except during the postnatal developmental period. Structures resembling granules and climbing fibers in Purkinje cells were deeply stained. In dentate neurons, long processes were deeply stained by anti-myosin Va, as were punctate nuclear structures. During the first postnatal year, myosin Va was differentially expressed in the external granular layer (EGL). In the EGL, proliferating prospective granule cells were not stained by anti-myosin Va antibody. In contrast, premigratory granule cells in the EGL stained moderately. Granule cells exhibiting a migratory profile in the molecular layer were also moderately stained. In conclusion, neuronal myosin Va is developmentally regulated, and appears to be required for cerebellar function from early postnatal life to senescence.

Myosin Va is developmentally regulated and expressed in the human cerebellum from birth to old age

Myosin Va functions as a processive, actin-based motor molecule highly enriched in the nervous system, which transports and/or tethers organelles, vesicles, and mRNA and protein translation machinery. Mutation of myosin Va leads to Griscelli disease that is associated with severe neurological deficits and a short life span. Despite playing a critical role in development, the expression of myosin Va in the central nervous system throughout the human life span has not been reported. To address this issue, the cerebellar expression of myosin Va from newborns to elderly humans was studied by immunohistochemistry using an affinity-purified anti-myosin Va antibody. Myosin Va was expressed at all ages from the 10th postnatal day to the 98th year of life, in molecular, Purkinje and granular cerebellar layers. Cerebellar myosin Va expression did not differ essentially in localization or intensity from childhood to old age, except during the postnatal developmental period. Structures resembling granules and climbing fibers in Purkinje cells were deeply stained. In dentate neurons, long processes were deeply stained by anti-myosin Va, as were punctate nuclear structures. During the first postnatal year, myosin Va was differentially expressed in the external granular layer (EGL). In the EGL, proliferating prospective granule cells were not stained by anti-myosin Va antibody. In contrast, premigratory granule cells in the EGL stained moderately. Granule cells exhibiting a migratory profile in the molecular layer were also moderately stained. In conclusion, neuronal myosin Va is developmentally regulated, and appears to be required for cerebellar function from early postnatal life to senescence.

O associativismo como estratégia de melhoria de rentabilidade da atividade leiteira em Glória de Dourados - MS / The associativism as strategy of improve the profit margin of the dairy cattle livestock activity in Glória de Dourados - MS / El asociativismo como estratégia para mejorar la rentabilidad de la actividad lechera en la ciudad de glória de Dourados - MS

O agronegócio do leite passou por grandes transformações nas últimas décadas, saindo da exploração de subsistência para a profissionalização, com enfoque para a produção em escala, com qualidade, agregação de valor e industrialização de produtos diferenciados. Em Mato Grosso do Sul, há necessidade de políticas que impulsionem o setor de produção a buscar maiores índices de produção e produtividade , bem como incentivo à produtividade, bem como incentivo à agroindústria a fim de agregar valor ao produto. Esta pesquisa realizada na bacia leiteira de Glória de Dourados, MS, teve como foco o elo da cadeia produtiva do leite entre produtor/indústria, no qual o setor primário opera com dificuldades, principalmente na comercialização do produto,fato evidenciado por aspectos como a oligopolização do setor industrial, aliado ao baixo poder de negociação do produtor, em função de aspectos culturais, de organização e econômicos. Este trabalho com base no associativismo rural visa o estudo da forma de negociação do leite e os aspectos que possam contribuir para que o produtor de leite possa ter maior segurança e rentabilidade. Com base nos índices obtidos, concluiu-se que a diminuição das incertezas comerciais do produtor ocorre quando a base produtiva e comercial está consubstanciada no associativismo, uma vez que fortalece a pequena produção. O modelo de comercialização coletiva com contratos formais oferece maior segurança, tanto para o produtor como para a indústria. Desta forma, as relações de mercado ficam bem definidas e o equilíbrio entre os agentes deste elo da cadeia se consolida. Este instrumento de gestão da comercialização, além de promover o equilíbrio, proporciona maior estabilidade e possibilidade real de planejamento e permanência do produtor de leite na atividade.

The dairy agribusiness has been going through great changes in the last decades, leaving the standard of subsistence exploration to become a workman like performance, focusing on commercial scale production, with high quality, value aggregation and industrialization of differentiated products. In Mato Grosso do Sul there have been lack of political action to boost the productive sector to get high levels of production and productivity, as well as incentive to agroindustrialization in order to add value to the products in natura. This project, which has been taking place in the basin dairy of Glória de Dourados, MS, focuses on the link between the dairy farmer and the industry, where the primary sector goes through hard times, mainly with regard to the trading of the product, as a result of the oligopolization of the industrial sector combined with the low power level of negotiation, due to cultural, social and economic aspects. This project is based upon rural associativism and aims the study about the strategies to negotiation and the aspects. Which can contribute to increase the assurance and profi t margin of the dairy farmers. Based upon the indexes of the results that were obtained via this research, one concludes that the decrease of the dairy farmer ́s commercial uncertainty occurs when his commercial productive basis is consolidated on associativism, once it strengthens the small scale production. The paradigm of the collective trade based upon formal contracts assures more confi dence to the farmer as far as to the industry. So, the market relationship between the agents of this link of the chain becomes well defi ned and a state of equilibrium is established. This management tool, besides promoting market equilibrium, provides more stability and real possibility for planning and assures the permanence of the dairy farmer in the activity.

La actividad lechera, como agronegócio, ha pasado por grandes transformaciones en las últimas décadas, dejando de ser exploración de manutención, cambiándose para la profi sionalización del sector con énfasis para la producción en escala. Con calidad, agregación de valor e industrialización de productos diferenciados. En Mato Grosso do Sul, hay necesidad de políticas de desarrollo para el sector productivo, para alcanzar índices de producción y productividad, bien como incentivo a la agroindustrialización, como medio de agregación de valor al producto. Esta encuesta hecha en la bacia lechera de Glória de Dourados, MS, tuvo como énfasis, el elo de la cadena produtiva de la leche productor/industria, donde el sector productivo primário opera con difi cultades, sobretodo cuando se habla acerca de la comercialización del producto, hecho comprobado por aspectos como la oligopolización del complejo industrial, aliado al pequeño poder de negociación del productor, teniendo en cuenta los aspectos culturales, aspectos organizacionales y económicos. Esta encuesta cuya basis es el asociativismo rural, tiene como fi nalidad el estudio de la forma de negociación de la leche y los aspectos que puedan contribuir para que el productor de leche pueda tener mayor seguridad y rentabilidad. Basándose en los índices obtenidos, concluyose que la diminuición de las incertidumbres comerciales del productor ocurre, cuando su base productiva y comercial están apoyadas en el asociativismo, teniéndose en cuenta que fortalece le pequeña producción. El paradigma de la comercialización coletiva con contractos formales, ofrece mayor seguridad, tanto para el productor quanto para la industria. Así, las relacionesde mercado se quedan bien defi nidas y el equilibrio entre lo agentes deste elo de la cadena consolídase. Este instrumento de gestión de comercialización, además de promover el equilibrio, proporciona mayor estabilidad y real posibilidad de planifi cación y permanéncia del productor en la actividad de ganadería lechera

Sensitivity to camptothecin in Aspergillus nidulans identifies a novel gene, scaA+, related to the cellular DNA damage response.

The anti-cancer drug camptothecin targets eukaryotic DNA topoisomerase I by trapping the covalent complex formed between the catalytically active enzyme and DNA. We are interested in identifying factors, other than topoisomerase I, that are involved in mediating cellular sensitivity to camptothecin. To this end, we have isolated eighteen mutants that are sensitive to camptothecin (sca) in the filamentous fungus Aspergillus nidulans and characterised one of them, sca299. The mutant sca299 is hypersensitive to camptothecin, and sensitive to several different mutagenic agents and to actinomycin D. Using temperature-sensitive mutations in genes that are known to regulate the cell cycle, we showed that the camptothecin sensitivity of the mutant sca299 is not affected by a mitotic block. The abnormal nuclear morphology observed in the sca299 mutant strain suggests that the germlings might be undergoing mitosis in the presence of unrepaired DNA damage, which would result in mitotic catastrophe. The hypersensitivity of the sca299 mutant to camptothecin does not result from elevated levels of topoisomerase I mRNA or from alterations in enzyme activity. Using DNA-mediated complementation of the sca299 mutant phenotype, the scaA+ gene was cloned. This gene encodes a 594-amino acid product; moderate structural similarity suggests that the scaA gene product may be related to the human nibrin gene which encodes a product involved in DNA double-strand break repair. Strains disrupted in the scaA gene were sensitive to the anti-topoisomerase I agent berberine, the DNA crosslinking agents mitomycin C and cis-platinum, and also to t-butyl hydroperoxide, which is an inducer of oxidative stress.

Tagging of genes involved in multidrug resistance in Aspergillus nidulans.

We have used a plasmid containing the Neurospora crassa pyr4 gene to transform an Aspergillus nidulans pyrG89 mutant strain in the presence of Bam-HI, and isolated multidrug-sensitive mutants among the transformants. Using this approach, we hoped to identify genes whose products are important for drug resistance by analyzing gene disruptions that alter the drug sensitivity of the cell. About 1300 transformants isolated following transformation were screened for sensitivity to drugs or various stress agents with different and/or the same mechanism of action. Seventy-seven of these transformants showed sensitivity to at least one drug, while fourteen transformants showed a complex phenotype of sensitivity to different drugs. The pyr4 marker was shown to be tightly linked to the mutant phenotype in only 36% of the pleiotropic mutants analyzed in sexual crosses. Genetic crosses between our multidrug-sensitive transformants and cycloheximide-sensitive and imazalil-resistant mutants of A nidulans were performed to determine whether mutations were present at the same loci. We have shown that the gene imaD that confers resistance to imazalil may also be involved in cycloheximide and hygromycin sensitivity, since this mutation is allelic to scyB (mutant scy290). In addition, the cross between the transformant R223 and the imazalil-resistant mutant ima535 showed that both mutations are in the same complementation group, suggesting that the gene imaG could also be involved in cycloheximide and itraconazole sensitivity.

Detection of Leptospira DNA in patients with aseptic meningitis by PCR.

Samples of cerebrospinal fluid from 103 patients with aseptic meningitis were tested by PCR for detection of leptospires, and the results were compared with those of the microscopic agglutination test (MAT) and an enzyme-linked immunosorbent assay for detection of immunoglobulin M (ELISA-IgM). Of these samples, 39.80% were positive by PCR and 8.74 and 3.88% were positive by MAT and ELISA-IgM, respectively.

Isolation and characterisation of cycloheximide-sensitive mutants of Aspergillus nidulans.

Aspergillus nidulans is a non-pathogenic fungus with well-developed genetics which provides an excellent model system for studying different aspects of drug resistance in filamentous fungi. As a preliminary step to characterizing genes that confer pleiotropic drug resistance in Aspergillus, we isolated cycloheximide-sensitive mutants of A. nidulans, which is normally resistant to this drug. The rationale for this approach is to identify genes whose products are important for drug resistance by analysing mutations that alter the resistance/sensitivity status of the cell. Fifteen cycloheximide-sensitive (named scy for sensitive to cycloheximide) mutants of A. nidulans were isolated and genetically characterised. Each scy mutant was crossed with the wild-type strain and five of the crosses gave 50% cycloheximide-sensitive progeny suggesting that they carry a single mutation required for cycloheximide sensitivity. We examined ten scy mutants for resistance/sensitivity to other drugs or stress agents with different and/or the same mechanism of action. Six of these mutants exhibited other altered resistance/sensitivity phenotypes which were linked to the cycloheximide sensitivity. These six mutants were analyzed by pairwise crosses and found to represent six linkage groups, named scyA-F. One of the mutants showed fragmentation of its vacuolar system and, in addition, its growth was osmotic, low-pH, and oxidative-stress sensitive.

Dot-ELISA for human leptospirosis employing immunodominant antigen.

A dot enzyme-linked immunosorbent assay which uses a proteinase-K resistant antigen (PK-Dot-ELISA) to detect antileptospiral IgM antibodies was compared to the microscopic agglutination test (MAT). The assay was evaluated in serum samples from patients with leptospirosis (n = 89), typhoid fever (n = 10), malaria (n = 19), syphilis (n = 20), hepatitis (n = 16) and clinically healthy individuals (n = 92). The PK-Dot-ELISA presented a sensitivity of 92.1% and a specificity of 97.5%. The overall results of the PK-Dot-ELISA were similar to those of the MAT. However, the PK-Dot-ELISA was capable of detecting antibody activity in 43% of acute-phase sera which were negative by the MAT. Our data suggest that PK-Dot-ELISA can be used as an important portable field serodiagnostic assay for acute leptospirosis.