Establishing a Robust In Vitro Embryonic Stem Cell Differentiation Assay to Monitor the Hematopoietic Potential of DELES Clones

Afin d’effectuer des études fonctionnelles sur le génome de la souris, notre laboratoire a généré une bibliothèque de clones de cellules souches embryonnaires (ESC) présentant des suppressions chromosomiques chevauchantes aléatoires – la bibliothèque DELES. Cette bibliothèque contient des délétions couvrant environ 25% du génome murin. Dans le laboratoire, nous comptons identifier de nouveaux déterminants du destin des cellules hématopoïétiques en utilisant cet outil. Un crible primaire utilisant la benzidine pour démontrer la présence d'hémoglobine dans des corps embryoïdes (EBS) a permis d’identifier plusieurs clones délétés présentant un phénotype hématopoïétique anormal. Comme cet essai ne vérifie que la présence d'hémoglobine, le but de mon projet est d'établir un essai in vitro de différenciation des ESC permettant de mesurer le potentiel hématopoïétique de clones DELES. Mon hypothèse est que l’essai de différenciation hématopoïétique publié par le Dr Keller peut être importé dans notre laboratoire et utilisé pour étudier l'engagement hématopoïétique des clones DELES. À l’aide d’essais de RT-QPCR et de FACS, j’ai pu contrôler la cinétique de différenciation hématopoïétique en suivant l’expression des gènes hématopoïétiques et des marqueurs de surface comme CD41, c-kit, RUNX1, GATA2, CD45, β-globine 1 et TER-119. Cet essai sera utilisé pour valider le potentiel hématopoïétique des clones DELES candidats identifiés dans le crible principal. Mon projet secondaire vise à utiliser la même stratégie rétro-virale a base de Cre-loxP utilisée pour générer la bibliothèque DELES pour générer une bibliothèque de cellules KBM-7 contenant des suppressions chromosomiques chevauchantes. Mon but ici est de tester si la lignée cellulaire leuémique humaine presque haploïde KBM-7 peut être exploitée en utilisant l'approche DELES pour créer cette bibliothèque. La bibliothèque de clones KBM-7 servira à définir les activités moléculaires de drogues anti-leucémiques potentielless que nous avons identifiées dans le laboratoire parce qu’elles inhibent la croissance cellulaire dans plusieurs échantillons de leucémie myéloïde aiguë dérivés de patients. Elle me permettra également d'identifier les voies de signalisation moléculaires qui, lorsque génétiquement perturbées, peuvent conférer une résistance à ces drogues.

A Rapid FACS-Based Strategy to Isolate Human Gene Knockin and Knockout Clones

Gene targeting protocols for mammalian cells remain inefficient and labor intensive. Here we describe FASTarget, a rapid, fluorescent cell sorting based strategy to isolate rare gene targeting events in human somatic cells. A fluorescent protein is used as a means for direct selection of targeted clones obviating the need for selection and outgrowth of drug resistant clones. Importantly, the use of a promoter-less, ATG-less construct greatly facilitates the recovery of correctly targeted cells. Using this method we report successful gene targeting in up to 94% of recovered human somatic cell clones. We create functional EYFP-tagged knockin clones in both transformed and non-transformed human somatic cell lines providing a valuable tool for mammalian cell biology. We further demonstrate the use of this technology to create gene knockouts. Using this generally applicable strategy we can recover gene targeted clones within approximately one month from DNA construct delivery to obtaining targeted monoclonal cell lines.

Descriptive catalog of Hemerocallis clones, 1893-1948,

"Sponsored by the American Plant Life Society and the Hemerocallis Society."

Incident Piza--Rio Branco. Grave situation politique au Brésil ...

An explanation of the author's reasons for resigning his position as Brazilian minister to France.

Barão do Rio Branco, notas politicas e biographicas

Originally published as a series of articles in "A Republica."

O Melro branco

Mode of access: Internet.

Camillo Castello Branco; typos e episodios da sua galeria.

Mode of access: Internet.

Nosographia de camillo castello branco.

Thesis (doctoral)--

De Monroe a Rio-Branco (pafinad de diplomaica) americana).

Entre George Canning e James Monroe.--A Assembléa do Isthmo.--A primeira Conferencia de Lima.--A Assembléa de Buenos Aires.--Tentativas de uma codificação.--A American latina e a diplomacia do imperio.

Gingerol content of diploid and tetraploid clones of ginger (Zingiber officinale Roscoe)

Ginger (Zingiber officinale, Roscoe), a monocotyledonous, sterile cultigen, is widely used as a spice, flavoring agent, and herbal medicine. The pungency of fresh ginger is due to a series of homologous phenolic ketones of which [6]-gingerol is the major one. The gingerols are thermally unstable and can be converted to their corresponding shogaols, which are present in dried ginger, Fresh rhizomes of 17 clones of Australian ginger, including commercial cultivars and experimental tetraploid clones, were assayed by HPLC for gingerols and shogaols. [6]-Gingerol was identified as the major pungent phenolic compound in all samples, while [8]- and [10]-gingerol occurred in lower concentrations. One cultivar known as Jamaican contained the highest concentrations of all three gingerols and was the most pungent of the clones analyzed. Gingerols were stable in ethanolic solution over a 5-month period when stored at 4 degrees C. Shogaols were not identified in the extracts prepared from fresh rhizomes at ambient temperature, confirming that these compounds are not native constituents of fresh ginger, In contrast to previous findings, this study did not find significant differences in gingerol concentrations between the tetraploid clones and their parent diploid cultivar.

Essential oil composition of diploid and tetraploid clones of ginger (Zingiber officinale roscoe) grown in Australia

Ginger oil, obtained by steam distillation of the rhizome of Zingiber officinale Roscoe, is used in the beverage and fragrance industries. Ginger oil displays considerable compositional diversity, but is typically characterized by a high content of sesquiterpene hydrocarbons, including zingiberene, arcurcumene, beta-bisabolene, and beta-sesquiphellandrene. Australian ginger oil has a reputation for possessing a particular lemony aroma, due to its high content of the isomers neral and geranial, often collectively referred to as citral. Fresh rhizomes of 17 clones of Australian ginger, including commercial cultivars and experimental tetraploid clones, were steam distilled 7 weeks post-harvest, and the resulting oils were analyzed by GC-MS. The essential oils of 16 of the 17 clones, including the tetraploid clones and their parent cultivar, were found to be of substantially similar composition. These oils were characterized by very high citral levels (51-71%) and relatively low levels of the sesquiterpene hydrocarbons typical of ginger oil. The citral levels of most of these oils exceeded those previously reported for ginger oils. The neral-to-geranial ratio was shown to be remarkably constant (0.61 +/- 0.01) across all 17 clones. One clone, the cultivar Jamaican, yielded oil with a substantially different composition, lower citral content and higher levels of sesquiterpene hydrocarbons. Because this cultivar also contains significantly higher concentrations of pungent gingerols, it possesses unique aroma and flavor characteristics, which should be of commercial interest.

CARLOS CASTELLO BRANCO: o comentarista paradigmático da imprensa brasileira

Carlos Castello Branco atuou nos principais jornais do país, vivendo principalmente em Brasília, de onde escreveu, durante 30 anos, a Coluna do Castello, espaço emblemático do jornalismo opinativo no século XX. Esta dissertação busca descrever as características do formato comentário, tendo como base a teoria dos gêneros jornalísticos. A pesquisa identificou oito tipos de comentários no jornalismo praticado pelo repórter piauiense, possibilitando a diferenciação das características relacionadas ao contexto histórico e à conjuntura política.

An outbreak of Serratia marcescens on the neonatal unit:a tale of two clones

Serratia spp. are an important cause of hospital-acquired infections and outbreaks in high-risk settings. Twenty-one patients were infected or colonized over a nine-month period during 2001-2002 on a neonatal unit. Twenty-two isolates collected were examined for antibiotic susceptibility, β-lactamase production and genotype. Random-amplified polymorphic DNA polymerase chain reaction and pulsed-field gel electrophoresis revealed that two clones were present. The first clone caused invasive clinical infection in four babies, and was subsequently replaced by a non-invasive clone that affected 14 babies. Phenotypically, the two strains also differed in their prodigiosin production; the first strain was non-pigmented whereas the second strain displayed pink-red pigmentation. Clinical features suggested a difference in their pathogenicity. No environmental source was found. The outbreak terminated following enhanced compliance with infection control measures and a change of antibiotic policy. Although S. marcescens continued to be isolated occasionally for another five months of follow-up, these were sporadic isolates with distinct molecular typing patterns. © 2005 The Hospital Infection Society.

Território para viver: dinâmicas territoriais da comunidade quilombola de Acauã, Poço Branco, Rio Grande do Norte

In the late 1980s, the quilombola (or maroon) communities emerged on the Brazilian public scene. They established themselves as new collective subjects and ethnic groups, in a historical moment of sensitive political changes in several social conflicts and struggles, both in Brazil and in Latin America. Because of their socio-cultural and historical singularities, these communities have self-identified in the same collective expression and have organized in search of recognition and respect for their rights. Quilombo communities and other self-labeled as "traditional communities" seek to reaffirm their differences in opposition to a conscious colonizer cultural project and re-signify their memories and traditions, that serve as reference in the construction of alternative production projects and community organization. One of the distinguishing characteristics of this quilombola political emergence process is the territorial nature of the struggles, manifested in at least two directions: on the one hand, the struggle for legal and formal recognition of a given space, i.e., the regularization and titling of occupied territories, considering that the Brazilian Constitution of 1988 recognizes the right of these communities to the final possession of the traditional lands. On the other hand, the struggle for recognition of their territoriality in a broader sense, not necessarily restricted to the demarcated area, but as the recognition of a culture and its own way of life, that originated historically in these territories. The current accomplishments and challenges of the Brazilian quilombola communities are well exemplified by the quilombo of Acauã, in the Poço Branco municipality of Rio Grande do Norte. The last fifteen years have been marked by important changes in this community, which has gained visibility and has emerged as a new political player. Acauã identified itself as quilombola community in 2004, the same year that it formalized its political structure, through the creation of the Association of Residents of Quilombo Acauã (AMQA, in Portuguese). Also in 2004, it requested to the National Institute of Colonization and Land Reform (INCRA, in Portuguese) the opening of the process for regularization and titling of quilombo territory, which is at an advanced stage, but so far without definitive resolution. This study aims to understand the process of territorialization (struggle for territorial claim) played in the last fifteen years by the community of Acauã.

Efeito da densidade de sementeira nos componentes de rendimento do tremoço branco (Lupinus albus L.)

O tremoço branco (Lupinus albus L.) é utilizado na alimentação animal e humana. Com este trabalho pretendeu-se avaliar o efeito de duas densidades de sementeira (20 e 40 sementes/m2, respectivamente) na produtividade e nos componentes do rendimento de um ecótipo de tremoço branco. O campo experimental foi instalado na Quinta do Galinheiro (ESAS) Santarém, de acordo com um delineamento experimental em parcelas totalmente aleatórias, com 4 repetições. Foram efetuadas observações bissemanais do desenvolvimento e crescimento através da amostragem aleatória de 8 plantas em cada um dos tratamentos (número de folhas, ramificações e vagens por planta; número médio de grãos por vagem e por planta; peso seco de cada componente e o total da planta; peso de 1000 grãos e teor de proteína do grão). Observaram-se perdas importantes de plantas entre a sementeira e a emergência. O maior crescimento e produtividade individual das plantas na menor densidade, relativamente à maior, justificaram os valores muito aproximados verificados entre os dois tratamentos quando são consideradas as variáveis por unidade de área. Os teores de proteína e calibre do grão foram também idênticos nos dois tratamentos. Confirma-se assim a elevada capacidade da planta em se adaptar a diferentes densidades.