Quinolinotriazole antiplasmodials via click chemistry: synthesis and in vitro studies of 7-Chloroquinoline-based compounds

Malaria is nowadays one of the most serious health concerns in a global scale and, although there is an evident increase in research studies in this area, pointed by the vast number of hits and leads, it still appears as a recurrent topic every year due to the drug resistance shown by the parasite exposing the urgent need to develop new antimalarial medications. In this work, 38 molecules were synthesized via copper(I)-catalyzed alkyne-azide cycloaddition (CuAAC) or "click" chemistry, following different routes to produce 2 different organic azides, obtained from a 4,7 dicholoquinoline, reacted with 19 different commercially available terminal alkynes. All those new compounds were evaluated for their in vitro activity against the chloroquine resistant malaria parasite Plasmodium falciparum (W2). The cytotoxicity evaluation was accomplished using Hep G2 cells and SI index was calculated for every molecule. Some of the quinoline derivatives have shown high antimalarial activity, with IC50 values in the range of 1.72-8.66 µM, low cytotoxicity, with CC50>1000 µM and selectivity index (SI) in the range of 20-100, with some compounds showing SI>800. Therefore, the quinolinotriazole hybrids could be considered a very important step on the development of new antimalarial drugs

Hacia una validación del territorio híbrido neuropsiquiátrico / Towards a validation of the hybrid neuropsychiatrie territory

El siglo XX presenció movimientos pendulares epistemológicos que guiaron la praxis psiquiátrica, tanto desde extremos de reduccionismo psicológico hasta reduccionismos biológicos. Han pasado ya más de treinta años desde el apogeo de la psiquiatría dinámica, especializada en la búsqueda de una hermenéutica de los síntomas mentales; y ahora, bien entrados en el siglo XXI atrás queda también la llamada «década del cerebro¼ que vio surgir una avalancha de conocimiento neurocientífico y psicofarmacológico que cambió la comprensión de las bases biológicas de los síntomas mentales. Por su parte, académicos como Germán Berrios han difundido el modelo de síntomas mentales de la escuela de Cambridge, que impresiona por su refinamiento y capacidad de síntesis: los síntomas mentales serían constructos híbridos, con núcleos de disfunción neurobiológica que son recubiertos, en mayor o menor grado, por un caparazón hermenéutico que no puede dejar de lado la vivencia subjetiva del síntoma ni la estructura de personalidad o el contexto histórico-cultural en el que toma lugar.

The 20th century witnessed epistemological pendulum movements that guided psychiatric praxis, both from extremes of psychological reductionism to biological reductionism. More than thirty years have passed since the heyday of dynamic psychiatry, specialized in the search for a hermeneutics of mental symptoms; and now, well into the 21st century, there is also the so-called "decade of the brain" that saw the emergence of an avalanche of neuroscientific and psychopharmacological knowledge that changed the understanding of the biological bases of mental symptoms. For their part, academics such as Germán Berrios have disseminated the Cambridge school model of mental symptoms, which impresses with its refinement and capacity for synthesis: mental symptoms would be hybrid constructs, with nuclei of neurobiological dysfunction that are covered, to a greater or lesser extent. to a lesser degree, due to a hermeneutical shell that cannot ignore the subjective experience of the symptom or the personality structure or the historical-cultural context in which it takes place.

Identification of the MYST3-CREBBP fusion gene in infants with acute myeloid leukemia and hemophagocytosis

ABSTRACT Background: Acute myeloid leukemia presenting the MYST3-CREBBP fusion gene is a rare subgroup associated with hemophagocytosis in early infancy and monocytic differentiation. The aim of this study was to define the relevant molecular cytogenetic characteristics of a unique series of early infancy acute myeloid leukemia cases (≤24 months old), based on the presence of hemophagocytosis by blast cells at diagnosis. Methods: A series of 266 infant cases of acute myeloid leukemia was the reference cohort for the present analysis. Acute myeloid leukemia cases with hemophagocytosis by blast cells were reviewed to investigate the presence of the MYST3-CREBBP fusion gene by fluorescence in situ hybridization (FISH) and reverse transcription polymerase chain reaction. Results: Eleven cases with hemophagocytosis were identified with hemophagocytic lymphohistiocytosis being ruled out. Six cases were classified as myelomonocytic leukemia, three as AML-M7 and two as AML-M2. In five cases, the presence of the MYST3-CREBBP fusion gene identified by molecular cytogenetics was confirmed by fluorescence in situ hybridization. All patients received treatment according to the Berlin-Frankfürt-Münster acute myeloid leukemia protocols and only one out of the five patients with the MYST3-CREBBP fusion gene is still alive. Conclusions: Our findings demonstrate that the presence of hemophagocytosis in acute myeloid leukemia was not exclusively associated to the MYST3-CREBBP fusion gene. Improvements in molecular cytogenetics may help to elucidate more complex chromosomal rearrangements in infants with acute myeloid leukemia and hemophagocytosis.

Parasitological assessment in the hybrid surubim (Pseudoplatystoma reticulatum x P. corruscans), with uncommon occurrence of Monogenea parasites / Avaliação parasitológica no híbrido surubim (Pseudoplatystoma reticulatum x P. corruscans), com ocorrência incomum de parasitos Monogenea

Abstract This study evaluated the parasite fauna of farmed hybrid surubim (Pseudoplatystoma reticulatum x P. corruscans) and the host-parasite-environment relationship in two fish farms located in Mato Grosso do Sul, Central Brazil, South America. A total of 120 hybrids from two different farms, 60 in each season (30 in the hot and 30 in cold season) were examined during a year. Water quality was weekly measured to evaluate the interaction among environmental conditions and parasitism. Histopathology was used to observe the effects of the parasites and environment on the fish gills. The ciliate protozoan Ichthyophthirius multifiliis and the monogeneans (Ameloblastella sp., Amphocleithrium paraguayensis, Vancleaveus ciccinus, V. fungulus and V. janacauensis) were the most prevalent parasites detected in both seasons in both farms, with prevalence above 80%. It was stated that parasites did not cause important damage in the health status of the hybrid surubim. These results might be related to general good management practices and environmental quality implemented by the fish farmers. The presence of uncommon monogenean parasites to this hybrid compared to their parents causing an environmental and ecological concern is here discussed.

Resumo Este estudo avaliou a fauna de parasitos do surubim híbrido cultivado (Pseudoplatystoma reticulatum x P. corruscans) e a relação hospedeiro-parasito-ambiente em duas pisciculturas localizadas no Estado do Mato Grosso do Sul, região Centro-Oeste, Brasil. Um total de 120 híbridos de duas fazendas, 60 em cada estação (30 na estação quente e 30 na fria), foram examinados durante um ano. A qualidade da água foi medida semanalmente para avaliar a interação entre as condições ambientais e o parasitismo. Histopatologia foi usada para observar os efeitos dos parasitos e do ambiente nas brânquias dos peixes. O protozoário ciliado Ichthyophthirius multifiliis e Monogenea (Ameloblastella sp., Amphocleithrium paraguayensis, Vancleaveus ciccinus, V. fungulus e V. janacauensis) foram os parasitos mais prevalentes detectados em ambas estações nas duas fazendas, com prevalências acima de 80%. Observou-se que os parasitos não causaram danos ao estado de saúde do surubim híbrido. Esses resultados estão relacionados às boas práticas de manejo e qualidade ambiental implementada pelos produtores. É discutida a presença incomum de Monogenea para esse híbrido, comparado com seus progenitores, podendo causar preocupação ambiental e ecológica.

Caracterização do potencial diagnóstico de poliantígenos para detecção do Trypanosoma cruzi na fase crônica da doença de Chagas / Chracterization of the potencial diagnostic of polyantigens for detecting Trypanosoma cruzi in the chronic phase of Chagas disease

O diagnóstico da doença de Chagas crônica (DCC) baseia-se em metodologias que usam em sua fase sólida antígenos brutos, semipurificados ou recombinantes, podendo resultar em baixa sensibilidade ou reação cruzada. Uma forma para resolver este problema é o uso de quimeras formadas por epítopos conservados e repetitivos de diferentes estruturas parasitárias em uma única molécula. O nosso objetivo foi caracterizar e avaliar o uso de quimeras em imunoensaios para o diagnóstico da DCC. As quimeras, IBMP-8.1, IBMP-8.2, IBMP-8.3 e IBMP-8.4 foram purificadas por meio de cromatografia e sua pureza avaliada por SDS-PAGE. Ensaios de dicroísmo circular (DC) e espalhamento dinâmico da luz (EDL) foram usados para avaliação do raio hidrodinâmico das quimeras, avaliação de sua estabilidade e escolha do sistema tampão que oferecesse o menor estado de agregação molecular. Ensaios sorológicos para detecção de anticorpos anti-Trypanosoma cruzi através de ELISA foram realizados utilizando um painel de 857 amostras positivas para a DCC e 689 negativas. Para avaliação de reação cruzada foram usadas 1079 amostras de diversas doenças endêmicas no país. A purificação foi eficiente uma vez que o SDS-PAGE indicou ausência de degradação das quimeras. As análises de DC e EDL mostraram que as quatro quimeras apresentaram menor tendência de agregação em tampão carbonato pH 9,6, sendo, assim, este o sistema para a sensibilização das placas. Os ensaios sorológicos revelaram valores elevados de sensibilidade (Sen) e especificidade (Esp) para a molécula IBMP-8.4 (Sen-99,3 por cento; Esp-100 por cento). O desempenho para as demais moléculas foi satisfatório, ficando os valores de Sen e Esp acima de 94 por cento. As moléculas IBMP-8.1, IBMP-8.2, IBMP-8.3 e IBMP-8.4 apresentaram respectivamente 0,46 por cento, 0,85 por cento, 0,46 por cento e 0,37 por cento de reação cruzada. Os resultados apontam que as quimeras atingiram os critérios de proficiência do Ministério da Saúde podendo, dessa forma, ser utilizados em ensaios diagnósticos para a DCC.

Establishment of a pheasant (Phasianus colchicus) spermatogonial stem cell line for the production of interspecies germ line chimeras

Background Spermatogonial stem cells (SSCs) are important for the production of interspecies germ line chimeras. The interspecies germ cell transfer technique has been suggested as a way to conserve endangered birds. Our objective was to develop a technique for restoring endangered birds by developing interspecies germ line chimeras between pheasant (Phasianus colchicus) and chicken (Gallus gallus) with SSCs. Results SSCs were isolated from the surgically removed testis of a pheasant. Growth conditions for pheasant SSCs were established by co-culturing STO (SIM mouse embryo-derived thioguanine and ouabain resistant) cells and pheasant SSCs. The colony-forming cells divided and proliferated stably to yield an established SSC line. Pheasant SSCs showed strong reactivity for GDNF family receptor alpha1 (GFRa1) marker. Finally, production of germ line chimeras was attempted by transferring pheasant SSCs into recipient embryos. Although final embryo survival was 5.6% (20/354), the initial survival rate was 88% (312/354). To measure the percent transfer of donor SSC to gonads, the pheasant SSCs were labeled with PKH 26 fluorescent dye. We observed 30% donor cells and 9.48% c-kit/CD117-positive cells in the gonads of recipient chickens. Donor SSCs were thus stably engrafted in the recipient gonads. Conclusions This study showed that SSCs can be used as a tool for the conservation of endangered birds and the production of germ line chimeras. Our findings yield insights into how we may use the pheasant spermatogonial stem cell line for efficient production of interspecies germ line chimeras and ultimately, to the restoration of endangered birds.

Phenotypic stability of hybrids of Gália melon in Rio Grande do Norte state, Brazil

The objectives of this study were to determine the importance of simple and complex components of the interaction genotype × environment and to evaluate the adaptability and stability of Gália melon hybrids. Nine hybrids were tested in twelve environments of Rio Grande Norte State from 2000 to 2001. The experiments were carried out in a randomized complete block design with three replications. The statistical methods of Toler and Burrows, Wricke and AMMI (Additive Main effect and Multiplicative Interaction) were used to study the adaptability and stability. The complex component is responsible for most of the genotype × environment interaction for the yield and content of solids soluble of fruits. The environments associated with Mossoró and Assu municipalities are the most suitable to evaluate melon hybrids in the state. The hybrid DRG 1537 was the most likely to be grown in the Agro-industrial Complex Mossoró-Assu due to its stability, high productivity and high content of soluble solids.

Os objetivos deste estudo foram determinar a importância das componentes simples e complexa da interação genótipo × ambiente e avaliar a adaptabilidade e estabilidade de híbridos de melão Gália. Nove híbridos foram testados em doze ambientes do Estado do Rio Grande Norte no período de 2000 a2001. Os experimentos foram conduzidos em blocos completos casualizados com três repetições. Os métodos estatísticos de Toler e Burrows, Wricke e AMMI (Additive Main effect and Multiplicative Interaction) foram usados para estudar a adaptabilidade e estabilidade. A componente complexa é responsável pela maior parte da interação genótipo × ambiente para a produtividade e teor de sólidos solúveis dos frutos. Os ambientes associados com Mossoró e Assu são os mais adequados para a avaliação de melão híbrido. O híbrido DRG1537 é o mais promissor para o cultivo no Complexo Agro-industrial Mossoró-Assu, devido à sua estabilidade, alta produtividade e alto teor de sólidos solúveis.

Hybridism between Biomphalaria cousini and Biomphalaria amazonica and its susceptibility to Schistosoma mansoni

Molecular techniques can aid in the classification of Biomphalaria species because morphological differentiation between these species is difficult. Previous studies using phylogeny, morphological and molecular taxonomy showed that some populations studied were Biomphalaria cousini instead of Biomphalaria amazonica. Three different molecular profiles were observed that enabled the separation of B. amazonica from B. cousini. The third profile showed an association between the two and suggested the possibility of hybrids between them. Therefore, the aim of this work was to investigate the hybridism between B. cousini and B. amazonica and to verify if the hybrids are susceptible to Schistosoma mansoni. Crosses using the albinism factor as a genetic marker were performed, with pigmented B. cousini and albino B. amazonica snails identified by polymerase chain reaction-restriction fragment length polymorphism. This procedure was conducted using B. cousini and B. amazonica of the type locality accordingly to Paraense, 1966. In addition, susceptibility studies were performed using snails obtained from the crosses (hybrids) and three S. mansoni strains (LE, SJ, AL). The crosses between B. amazonica and B. cousini confirmed the occurrence of hybrids. Moreover, hybrids can be considered potential hosts of S. mansoni because they are susceptible to LE, SJ and AL strains (4.4 percent, 5.6 percent and 2.2 percent, respectively). These results indicate that there is a risk of introducing schistosomiasis mansoni into new areas.

Edad y crecimiento del híbrido de tilapia Oreochromis niloticus x Oreochromis aureus (Perciformes: Cichlidae) en la represa “Zimapán” Hidalgo, México

Age and growth of the hybrid tilapia Oreochromis niloticus x Oreochromis aureus (Perciformes: Cichlidae) in the dam “Zimapan” Mexico. Studies on age and growth in fishes are essential to establish models on population dynamics. The previos issues were determined in this study by scale growth analyses. Between September 2003 and May 2004, 382 organisms were captured using gill and atarraya nets. Eight samples were developed every month a long this study. Standard length in centimeters (Le), total weight in grams (Pt), scale length in centimeters (Lesc) and growth rings were determined. The size and weight ranged from 38 to 232mm (Le) and 8.7 to 311.9g (Pt), respectively. The growth rings analyses reveal four age groups, being the sec-ond group the most representative with 34.3% of the total sampled. The scale ring analyses showed two growth rings, one develops between November-January and the other between April-May, in good correlation with low temperature and gonad development, respectively. The growth parameters were established as: L∞=281.1mm, W∞=877.1g, K=0.33 and T0=0.88year -1. The growth curves based on length and weight were described by using the von Bertalanffy model: Le =28.11[1-e-0.33(t+0.88)] and Pt =877.17[1-e-0.33(t+0.88)]³. Finally, it is concluded that commercial catches in this dam are between a year and year and half-old when exploited. Rev. Biol. Trop. 59 (2): 761-770. Epub 2011 June 01.

La edad y crecimiento de la tilapia Oreochromis niloticus x Oreochromis aureus fueron determinados por medio de la lectura de escamas. Un total de 382 individuos fueron capturados mediante el uso de redes agalleras y atarrayas. Ocho recolectas mensuales se llevaron a cabo entre septiembre 2003-mayo 2004; se registraron los datos de longitud estándar en cm. (Le), el peso total en g (Pt) y se obtuvieron muestras de escamas para registrar el largo de la escama en mm. (Lesc) y el número de anillos de crecimiento formados. Las tallas y pesos de recolecta variaron entre 38 a 232mm (Le) y de 8.7 a 311.9g de peso total (Pt) respectivamente. El recuento de los anillos anuales en escamas, permitió establecer cuatro grupos de edad, siendo el grupo de edad 2 el más representativo, con un 34.3% de la población muestreada. El cálculo de los incrementos marginales del crecimiento mensual de las escamas, sugirió la formación de dos anillos, uno de ellos se forma en los meses de noviembre-enero y otro en los meses de abril-mayo, los cuales coinciden con la época de bajas temperaturas y de madurez gonádica respectivamente. Los parámetros de crecimiento quedaron establecidos en: L∞=281.1mm, W∞=877.1g, K=0.33 y T0=0.88/año. Las curvas de crecimiento en longitud y peso se describieron mediante las ecuaciones de von Bertalanffy: Le=28.11 [1- e-0.33(t+0.88)] y Pt=877.17 [1-e-0.33(t+0.88)]³. Finalmente, se concluyó que los peces que se explotan en la represa mediante captura comercial presentan entre un año y año y medio de edad.

Caracterización del perfil de ácidos grasos en granos de híbridos de maíz blanco cultivados en Venezuela / Fatty acids profile characterization of white maize hybrids grown in Venezuela

En Venezuela el maíz blanco representa el principal cultivo por área de siembra, producción y consumo. Uno de los principales subproductos de este rubro es el aceite de maíz, cuyo efecto positivo sobre la salud, por su alta proporción de ácidos grasos insaturados, ha sido ampliamente reconocido. Con el fin de conocer el perfil de ácidos grasos de doce híbridos de maíz blanco sembrados extensivamente en Venezuela y el efecto que distintas localidades de siembra poseen sobre el mismo, se realizaron tres ensayos a escala semicomercial en tres localidades de los estados Portuguesa, Yaracuy y Guárico. Las proporciones de los principales ácidos grasos del aceite crudo de las muestras de grano de distintos híbridos fueron determinados usando cromatografía de gases. Se encontraron diferencias significativas (p<0,01) entre los híbridos para el contenido de los ácidos grasos: araquídico, palmítico, palmitoléico, esteárico, oleico, gadoléico y linoléico; no se encontró diferencias para el ácido linolénico. Adicionalmente, el efecto de las localidades fue altamente significativo para todos los ácidos grasos mencionados. Al correlacionar el contenido de estos ácidos grasos se encontraron relaciones altas y significativas; las relaciones más estrechas fueron: linoleico-oleico (Rho= -0,98**), araquídico-palmítico (Rho= -0,61**), linoleico-esteárico (Rho= -0,61**) y oleico-esteárico (Rho= 0,58**). Los maíces producidos en Venezuela presentaron menores niveles de ácido linoléico y mayores de ácidos palmítico, esteárico y oleico con respecto los maíces referenciados para climas templados. Estas diferencias implican cambios en las propiedades alimenticias del aceite de maíz proveniente de maíces sembrados en Venezuela y deben tomarse en cuenta en el desarrollo de nuevos cultivares y en el procesamiento industrial con fines de aprovechamiento de aceites.

In Venezuela, white corn is the most important crop regarding production, harvest area and consumption. One of its main by-products is corn oil, whose positive effect on health caused by the high content of unsaturated fatty acids has been widely recognized. In order to characterize the fatty acids profile of twelve white grained maize hybrids extensively grown in Venezuela, and the effect that divergent localities has on this profile, three semi commercial scale trials where established in Portuguesa, Yaracuy and Guárico states. Proportions of the main fatty acids in the raw oil of the different grain samples were determined using gas chromatography. Significant differences (p<0,01) between hybrids were found for arachidic, palmitic, stearic, oleic, gadoleic and linoleic acids; non significant differences were found for linolenic acid. Significant differences between localities were found for all the fatty acids evaluated. High and significant correlations between fatty acids content were found; the most important relations were: linoleic-oleic (Rho= -0,98**), arachidic-palmitic (Rho= -0,61**), linoleic-stearic (Rho= -0,61**) and oleic-stearic (Rho= 0,58**). Corn produced in Venezuela presents lower levels of linoleic and higher levels of palmitic, stearic and oleic acids than the levels found in temperate corn. These differences involve significant changes in the nutritional properties of Venezuelan corn oil that should be considered in the development of new cultivars and industrial processes for oil production.

Occurrence of hybrids and laboratory evidence of fertility among three species of the Phyllosoma complex (Hemiptera: Reduviidae) in Mexico

In seven studied communities of Western Mexico, triatomine specimens were sympatrically collected, some with atypical morphological characteristics in contrast to pure specimens, which were presumed to be hybrids. More than 200 specimens of Meccus pallidipennis and Meccus longipennis with brown-yellow markings on dorsal connexival segments were collected in Ahuacapán and Quitupan. In La Mesa, more than 60 specimens similar to Meccus picturatus in most morphological characteristics (including size) were collected, although they presented a largely yellowish corium like M. pallidipennis. Interfertility was proven between all of the studied wild hybrid specimens, as well as between all the experimental laboratory hybrids. Two different phenotypes (M. picturatus and M. longipennis) were obtained from crosses between M. picturatus x M. picturatus and M. longipennis x M. longipennis from the three studied localities in state of Nayarit as from La Mesita. Results support the hypothesis that the subspecific ranking of those triatomines may, therefore, be more appropriate because reproductive isolation has not been developed and complete interbreeding was recorded.

Characterization of the acute inflammatory response in the hybrid tambacu (Piaractus mesopotamicus male × Colossoma macropomum female) (Osteichthyes) / Caracterização da resposta inflamatória aguda no híbrido tambacu (Piaractus mesopotamicus macho × Colossoma macropomum fêmea) (Osteichthyes)

This work evaluated the acute inflammatory response induced by injections of 0.5 mL saline solution (control), 500 µg carrageenin and 0.5 mL thioglycollate 3 percent in the swim bladder of juvenile tambacu hybrid. Fish were distributed in three treatments, three replications and acclimated for a period of 10 days before assay. The cell characterization from the inflammatory exudate was performed in Giemsa and PAS stained smears. Carrageenin, injected in fish, showed an increase on the total number of cells in the inflammatory exudate when compared to saline and thioglycollate injected. Whereas, for carrageenin-injected fish, the percentage of thrombocyte was higher than thioglycollate. On the other hand, granulocyte percentage in thioglycollate-injected fish was higher than the ones injected using carrageenin. Carrageenin provoked the highest migration of macrophage to the inflammatory site. The PAS method confirmed the presence of three types of granulocytes: eosinophilic granular cell (EGC) type 1 with the characteristics of a special granulocytic cell commonly found in the circulating blood; EGC type 2 shorter than the last one and neutrophil. This study contributes to a better understanding of the inflammatory response and infectious processes in native fish.

Este estudo avaliou a resposta inflamatória aguda induzida por injeções de 0,5 mL de solução salina (controle), 500 µg de carragenina e 0,5 mL de tioglicolato a 3 por cento na bexiga natatória de juvenis do híbrido tambacu. Os peixes foram distribuídos em três tratamentos, três repetições e aclimatados durante 10 dias antes do ensaio. A caracterização das células do exsudato inflamatório foi feita após coloração com Giemsa e PAS. Peixes injetados com carragenina apresentaram maior número de células no exsudato inflamatório do que com salina e tioglicolato. A porcentagem de trombócitos no exsudato foi maior nos injetados com carragenina quando comparada com a dos injetados com tioglicolato. Por outro lado, o percentual de granulócitos foi maior em animais injetados com tioglicolato do que em animais injetados com carragenina. A carragenina provocou maior migração de macrófagos para o foco inflamatório. O método de PAS confirmou a presença de três tipos de granulócitos: célula granular eosinofílica (CGE) tipo 1 com as características da célula granulocítica especial encontrada no sangue, CGE tipo 2, menor do que esta última, e de neutrófilos. Este estudo contribui para o melhor entendimento da resposta inflamatória e dos processos infecciosos em peixes nativos.

Biological and genetic aspects of experimental hybrids from species of the Phyllosoma complex (Hemiptera: Reduviidae: Triatominae)

The present work is a thorough investigation of the degree of reproductive isolation between Meccus mazzottii and Meccus longipennis, Meccus picturatus, Meccus pallidipennis and Meccus bassolsae, as well as between M. longipennis and M. picturatus. We examined fertility and segregation of morphological characteristics in two generations of hybrids derived from crosses between these species. The percentage of pairs with (fertile) offspring was highest in the set of crosses between M. longipennis and M. picturatus, and lowest between M. mazzottii and M. picturatus. Most first-generation (F1) individuals from crosses involving M. mazzottii were morphologically similar to this species, while only F1 x F1 progeny of parental crosses between M. mazzottii and M. longipennis had offspring second generation that looked like M. mazzottii. The results indicate that different degrees of reproductive isolation apparently exist among the species of the Phyllosoma complex examined in this study. The biological evidence obtained in this study does not support the proposal that M. longipennis and M. picturatus are full species. It could indicate on the contrary, that both could be considered as subspecies of a single polytypic species. On the other hand, biological evidence supports the proposal that M. mazzottii is a full species.

Report of a chimeric origin of transposable elements in a bovine-coding gene

Despite the wide distribution of transposable elements (TEs) in mammalian genomes, part of their evolutionary significance remains to be discovered. Today there is a substantial amount of evidence showing that TEs are involved in the generation of new exons in different species. In the present study, we searched 22,805 genes and reported the occurrence of TE-cassettes in coding sequences of 542 cow genes using the RepeatMasker program. Despite the significant number (542) of genes with TE insertions in exons only 14 (2.6%) of them were translated into protein, which we characterized as chimeric genes. From these chimeric genes, only the FAST kinase domains 3 (FASTKD3) gene, present on chromosome BTA 20, is a functional gene and showed evidence of the exaptation event. The genome sequence analysis showed that the last exon coding sequence of bovine FASTKD3 is ~85% similar to the ART2A retrotransposon sequence. In addition, comparison among FASTKD3 proteins shows that the last exon is very divergent from those of Homo sapiens, Pan troglodytes and Canis familiares. We suggest that the gene structure of bovine FASTKD3 gene could have originated by several ectopic recombinations between TE copies. Additionally, the absence of TE sequences in all other species analyzed suggests that the TE insertion is clade-specific, mainly in the ruminant lineage.

Mesenchymal stem cells from patients with chronic myeloid leukemia do not express BCR-ABL and have absence of chimerism after allogeneic bone marrow transplant

Bone marrow is a heterogeneous cell population which includes hematopoietic and mesenchymal progenitor cells. Dysregulated hematopoiesis occurs in chronic myelogenous leukemia (CML), being caused at least in part by abnormalities in the hematopoietic progenitors. However, the role of mesenchymal stem cells (MSCs) in CML has not been well characterized. The objectives of the present study were to observe the biological characteristics of MSCs from CML patients and to determine if MSCs originate in part from donors in CML patients after bone marrow transplantation (BMT). We analyzed MSCs from 5 untreated patients and from 3 CML patients after sex-mismatched allogeneic BMT. Flow cytometry analysis revealed the typical MSC phenotype and in vitro assays showed ability to differentiate into adipocytes and osteoblasts. Moreover, although some RT-PCR data were contradictory, combined fluorescence in situ hybridization analysis showed that MSCs from CML patients do not express the bcr-abl gene. Regarding MSCs of donor origin, although it is possible to detect Y target sequence by nested PCR, the low frequency (0.14 and 0.34 percent) of XY cells in 2 MSC CML patients by fluorescence in situ hybridization analysis suggests the presence of contaminant hematopoietic cells and the absence of host-derived MSCs in CML patients. Therefore, we conclude that MSCs from CML patients express the typical MSC phenotype, can differentiate into osteogenic and adipogenic lineages and do not express the bcr-abl gene. MSCs cannot be found in recipients 12 to 20 months after BMT. The influence of MSCs on the dysregulation of hematopoiesis in CML patients deserves further investigation.

Cassava genetic resources and their utilization for breeding of the crop

Wild cassava relatives are perennials and vary in growth pattern from nearly acaulescent subshrubs to small trees. They have been used as a source of useful characters such as high protein content, apomixis, resistance to mealybug and mosaic disease, and tolerance to drought. Indigenous clones are a potential source of beta-carotene and lycopene. Apomixis genes have been transferred to the crop successfully through interspecific hybridization, and apomictic clones arising from these hybrids are now being grown at the Universidade de Brasília. Interspecific hybrids produced earlier were polyploidized and had their fertility restored. Different useful types of chimera were also produced.

The synthesis of a new cassava-derived species, Manihot vieiri Nassar

A new species was synthesized artificially by chromosome doubling in a hybrid. The ensuing polyploid type exhibits an apomictic nature and maintains its morphological characteristics in the progeny. It showed a frequency of multiembryonic sacs of 29% in the ovules examined, whereas sacs were absent in the diploid type.