Three linked enzyme loci in fishes: implications in the evolution of vertebrate chromosomes.

A three-point linkage group comprised of loci coding for adenosine deaminase (ADA), glucose-6-phosphate dehydrogenase (G6PDH), and 6-phospho-gluconate dehydrogenase (6PGD) is described in fish of the genus Xiphophorus (Poeciliidae). The alleles at loci in this group were shown to assort independently from the alleles at three other loci--isocitrate dehydrogenase 1 and 2, and glyceraldehyde-3-phosphate dehydrogenase 1. Alleles at the latter three loci also assort independently from each other. Data were obtained by observing the segregation of electrophoretically variant alleles in reciprocal backcross hybrids derived from crosses between either X. helleri guentheri or X. h. strigatus and X. maculatus. The linkage component of chi2 was significant (less than 0.01) in all crosses, indicating that the linkage group is conserved in all populations of both species of Xiphophorus examined. While data from X. h. guentheri backcrosses indicate the linkage relationship ADA--6%--G6PDH--24%--6PGD, and ADA--29%--6PGD (30% when corrected for double crossovers), data from backcrosses involving strigatus, while supporting the same gene order, yielded significantly different recombination frequencies. The likelihood of the difference being due to an inversion could not be separated from the possibility of a sex effect on recombination in the present data. The linkage of 6PGD and G6PDH has been shown to exist in species of at least three classes of vertebrates, indicating the possibility of evolutionary conservation of this linkage.

THE ROLE OF MUCOSAL EPITHELIAL CELLS IN HIV-1 INFECTION

The predominant route of human immunodeficiency virus type 1 (HIV-1) transmission is infection across the vaginal mucosa. Epithelial cells, which form the primary barrier of protection against pathogens, are the first cell type at these mucosal tissues to encounter the virus but their role in HIV infection has not been clearly elucidated. Although mucosal epithelial cells express only low levels of the receptors required for successful HIV infection, productive infection does occur at these sites. The present work provides evidence to show that HIV exposure, without the need for productive infection, induces human cervical epithelial cells to produce Thymic Stromal Lymphopoietin (TSLP), an IL7-like cytokine, which potently activated human myeloid dendritic cells (mDC) to cause the homeostatic proliferation of autologous CD4+ T cells that serve as targets for HIV infection. Rhesus macaques inoculated with simian immunodeficiency virus (SIV) or with the simian-human immunodeficiency virus (SHIV) by the vaginal, oral or rectal route exhibited dramatic increases in: TSLP expression, DC and CD4+ T cell numbers, and viral replication, in the vaginal, oral, and rectal tissues, respectively within the first 2 weeks after virus exposure. Evidence obtained showed that HIV-mediated TSLP production by cervical cells is dependent upon the expression of the cell surface salivary agglutinin (SAG) protein gp340. Epithelial cells expressing gp340 exhibited HIV endocytosis and TSLP expression and genetic knockdown of gp340 or use of a gp340-blocking antibody inhibited TSLP expression by HIV. On the other hand, gp340-null epithelial cells failed to endocytose HIV and produce TSLP, but transfection of gp340 resulted in HIV-induced TSLP expression. Finally, HIV-induced TSLP expression was found to be mediated by TLR7/8 signaling and NF-kB activity because silencing these pathways or use of specific inhibitors abrogated TSLP expression in gp340-postive but not in gp340-null epithelial cells. Overall these studies identify TSLP as a key player in the acute phase of HIV-1 infection in permitting HIV to successfully maneuver the hostile vaginal mucosal microenvironment by creating a conducive environment for sustaining the small amount of virus that initially crosses the mucosal barrier allowing it to successfully cause infection and spread to distal compartments of the body

MODELING SPORADIC TUMOR FORMATION DRIVEN BY TELOMERE DYSFUNCTION IN THE GASTROINTESTINAL TRACT

Colorectal cancer is a complex disease that is thought to arise when cells accumulate mutations that allow for uncontrolled growth. There are several recognized mechanisms for generating such mutations in sporadic colon cancer; one of which is chromosomal instability (CIN). One hypothesized driver of CIN in cancer is the improper repair of dysfunctional telomeres. Telomeres comprise the linear ends of chromosomes and play a dual role in cancer. Its length is maintained by the ribonucleoprotein, telomerase, which is not a normally expressed in somatic cells and as cells divide, telomeres continuously shorten. Critically shortened telomeres are considered dysfunctional as they are recognized as sites of DNA damage and cells respond by entering into replicative senescence or apoptosis, a process that is p53-dependent and the mechanism for telomere-induced tumor suppression. Loss of this checkpoint and improper repair of dysfunctional telomeres can initiate a cycle of fusion, bridge and breakage that can lead to chromosomal changes and genomic instability, a process that can lead to transformation of normal cells to cancer cells. Mouse models of telomere dysfunction are currently based on knocking out the telomerase protein or RNA component; however, the naturally long telomeres of mice require multiple generational crosses of telomerase null mice to achieve critically short telomeres. Shelterin is a complex of six core proteins that bind to telomeres specifically. Pot1a is a highly conserved member of this complex that specifically binds to the telomeric single-stranded 3’ G-rich overhang. Previous work in our lab has shown that Pot1a is essential for chromosomal end protection as deletion of Pot1a in murine embryonic fibroblasts (MEFs) leads to open telomere ends that initiate a DNA damage response mediated by ATR, resulting in p53-dependent cellular senescence. Loss of Pot1a in the background of p53 deficiency results in increased aberrant homologous recombination at telomeres and elevated genomic instability, which allows Pot1a-/-, p53-/- MEFs to form tumors when injected into SCID mice. These phenotypes are similar to those seen in cells with critically shortened telomeres. In this work, we created a mouse model of telomere ysfunction in the gastrointestinal tract through the conditional deletion of Pot1a that recapitulates the microscopic features seen in severe telomere attrition. Combined intestinal loss of Pot1a and p53 lead to formation of invasive adenocarcinomas in the small and large intestines. The tumors formed with long latency, low multiplicity and had complex genomes due to chromosomal instability, features similar to those seen in sporadic human colorectal cancers. Taken together, we have developed a novel mouse model of intestinal tumorigenesis based on genomic instability driven by telomere dysfunction.

Phenotypic and genotypic analysis of mouse hepatitis virus (JHM) complementation group A temperature-sensitive mutants

Temperature sensitive (ts) mutant viruses have helped elucidate replication processes in many viral systems. Several panels of replication-defective ts mutants in which viral RNA synthesis is abolished at the nonpermissive temperature (RNA$\sp{-})$ have been isolated for Mouse Hepatitis Virus, MHV (Robb et al., 1979; Koolen et al., 1983; Martin et al., 1988; Schaad et al., 1990). However, no one had investigated genetic or phenotypic relationships between these different mutant panels. In order to determine how the panel of MHV-JHM RNA$\sp{-}$ ts mutants (Robb et al., 1979) were genetically related to other described MHV RNA$\sp{-}$ ts mutants, the MHV-JHM mutants were tested for complementation with representatives from two different sets of MHV-A59 ts mutants (Koolen et al., 1983; Schaad et al., 1990). The three ts mutant panels together were found to comprise eight genetically distinct complementation groups. Of these eight complementation groups, three complementation classes are unique to their particular mutant panel; genetically equivalent mutants were not observed within the other two mutant panels. Two complementation groups were common to all three mutant panels. The three remaining complementation groups overlapped two of the three mutant sets. Mutants MHV-JHM tsA204 and MHV-A59 ts261 were shown to be within one of these overlapping complementation groups. The phenotype of the MHV-JHM mutants within this complementation class has been previously characterized (Leibowitz et al., 1982; Leibowitz et al, 1990). When these mutants were grown at the permissive temperature, then shifted up to the nonpermissive temperature at the start of RNA synthesis, genome-length RNA and leader RNA fragments accumulated, but no subgenomic mRNA was synthesized. MHV-A59 ts261 produced leader RNA fragments identical to those observed with MHV-JHM tsA204. Thus, these two MHV RNA$\sp{-}$ ts mutants that were genetically equivalent by complementation testing were phenotypically similar as well. Recombination frequencies obtained from crosses of MHV-A59 ts261 with several of the gene 1 MHV-A59 mutants indicated that the causal mutation(s) of MHV-A59 ts261 was located near the overlapping junction of ORF1a and ORF1b, in the 3$\sp\prime$ end of ORF1a, or the 5$\sp\prime$ end of ORF1b. Sequence analysis of this junction and 1400 nucleotides into the 5$\sp\prime$ end of ORF1b of MHV-A59 ts261 revealed one nucleotide change from the wildtype MHV-A59. This substitution at nucleotide 13,598 (A to G) was a silent mutation in the ORF1a reading frame, but resulted in an amino acid change in ORF1b gene product (I to V). This amino acid change would be expressed only in the readthrough translation product produced upon successful ribosome frameshifting. A revertant of MHV-A59 ts261 (R2) also retained this guanidine residue, but had a second substitution at nucleotide 14,475 in ORF1b. This mutation results in the substitution of valine for an isoleucine.^ The data presented here suggest that the mutation in MHV-A59 ts261 (nucleotide 13,598) would be responsible for the MHV-JHM complementation group A phenotype. A second-site reversion at nucleotide 14,475 may correct this defect in the revertant. Sequencing of gene 1 immediately upstream of nucleotide 13,296 and downstream of nucleotide 15,010 must be conducted to test this hypothesis. ^

Archaeological Research at Sirkeli Höyük: Preliminary Results of the Latest Excavations (2012 & 2013) and their Implications

Sirkeli Höyük is an ancient settlement located 40 km east of Adana on the left bank of the Ceyhan River in Plain Cilicia. The main mound covers an area of approximately 300×400 m and rises to a height of ca. 30 m above the level of the surrounding plain. Due to its strategic location overlooking a road that crosses the Misis mountains, Sirkeli Höyük always played an important role within Plain Cilicia. J. Garstang’s (1936-1937), B. Hrouda’s (1992-1996) and H. Ehringhaus’ (1997) excavations have shown that the site was occupied from the 4th to late 1st millennium B.C. Since 2006, a new Swiss-Turkish team is investigating Sirkeli Höyük again. Due to modern excavation techniques and an interdisciplinary approach, the architectural and material remains that have been uncovered by the new excavations have yielded much new information. Apart from a more precise pottery sequence, the new project has discovered an extensive lower town surrounded by an elaborate double city wall. The paper will summarize the results that have been gathered since 2006, with particular focus on the campaigns 2012-2013, and aims to show how they may contribute to the understanding of the cultural developments in this region.

Disentangling the role of phenotypic plasticity and genetic divergence in contemporary ecotype formation during a biological invasion

The occurrence of contemporary ecotype formation through adaptive divergence of populations within the range of an invasive species typically requires standing genetic variation but can be facilitated by phenotypic plasticity. The relative contributions of both of these to adaptive trait differentiation have rarely been simultaneously quantified in recently diverging vertebrate populations. Here we study a case of intraspecific divergence into distinct lake and stream ecotypes of threespine stickleback that evolved in the past 140 years within the invasive range in Switzerland. Using a controlled laboratory experiment with full-sib crosses and treatments mimicking a key feature of ecotypic niche divergence, we test if the phenotypic divergence that we observe in the wild results from phenotypic plasticity or divergent genetic predisposition. Our experimental groups show qualitatively similar phenotypic divergence as those observed among wild adults. The relative contribution of plasticity and divergent genetic predisposition differs among the traits studied, with traits related to the biomechanics of feeding showing a stronger genetic predisposition, whereas traits related to locomotion are mainly plastic. These results implicate that phenotypic plasticity and standing genetic variation interacted during contemporary ecotype formation in this case.

Hybrid Breakdown in Cichlid Fish

Studies from a wide diversity of taxa have shown a negative relationship between genetic compatibility and the divergence time of hybridizing genomes. Theory predicts the main breakdown of fitness to happen after the F1 hybrid generation, when heterosis subsides and recessive allelic (Dobzhansky-Muller) incompatibilities are increasingly unmasked. We measured the fitness of F2 hybrids of African haplochromine cichlid fish bred from species pairs spanning several thousand to several million years divergence time. F2 hybrids consistently showed the lowest viability compared to F1 hybrids and non-hybrid crosses (crosses within the grandparental species), in agreement with hybrid breakdown. Especially the short- and long-term survival (2 weeks to 6 months) of F2 hybrids was significantly reduced. Overall, F2 hybrids showed a fitness reduction of 21% compared to F1 hybrids, and a reduction of 43% compared to the grandparental, non-hybrid crosses. We further observed a decrease of F2 hybrid viability with the genetic distance between grandparental lineages, suggesting an important role for negative epistatic interactions in cichlid fish postzygotic isolation. The estimated time window for successful production of F2 hybrids resulting from our data is consistent with the estimated divergence time between the multiple ancestral lineages that presumably hybridized in three major adaptive radiations of African cichlids.

Functional analysis of the Galpha protein, GNA -3, in the development of Neurospora crassa using biochemical and genetic studies

Extracellular signals regulate fungal development and, to sense and respond to these cues, fungi evolved signal transduction pathways similar to those in mammalian systems. In fungi, heterotrimeric G proteins, composed of α, β, and γ subunits, transduce many signals, such as pheromones and nutrients, intracellularly to alter adenylyl cyclase and MAPK cascades activity. ^ Previously, the Gα proteins GNA-1 and GNA-2 were characterized in regulating development in the fungus Neurospora crassa. R. A. Baasiri isolated a third Gα, gna-3, and P. S. Rowley generated Δgna-3 mutants. GNA-3 belongs to a fungal Gα family that regulates cAMP metabolism and virulence. The Δ gna-3 sexual cycle is defective in homozygous crosses, producing inviable spores. Δgna-3 mutants have reduced aerial hyphae formation and derepressed asexual sporulation (conidiation), causing accumulation of asexual spores (conidia). These defects are similar to an adenylyl cyclase mutant, cr-1; cAMP supplementation suppressed Δ gna-3 and cr-1. Inappropriate conidiation and expression of a conidiation gene, con-10, were higher in Δ gna-3 than cr-1 submerged cultures; peptone suppressed conidiation. Adenylyl cyclase activity and expression demonstrated that GNA-3 regulates enzyme levels. ^ A Δgna-1 cr-1 was analyzed with F. D. Ivey to differentiate GNA-1 roles in cAMP-dependent and -independent pathways. Δ gna-1 cr-1 defects were worse than cr-1 and refractory to cAMP, suggesting that GNA-1 is necessary for sensing extracellular CAMP. Submerged culture conidiation was highest in Δgna-1 cr-1, and only high cell density Δgna-1 cultures conidiated, which correlated with con-10 levels. Transcription of a putative heat shock cognate protein was highest in Δgna-1 cr-1. ^ Functional relationships between the three Gαs was analyzed by constructing Δgna-1 Δgna-2 Δ gna-3, Δgna-1 Δgna-3, and Δgna-2 Δgna-3 strains. Δ gna-2 Δgna-3 strains exhibited intensified Δ gna-3 phenotypes; Δgna-1 Δgna-2 Δgna-3 and Δgna-1 Δ gna-3 strains were identical to Δgna-1 cr-1 on plates and were non-responsive to cAMP. The highest levels of conidiation and con-10 were detected in submerged cultures of Δ gna-1 Δgna-2 Δgna-3 and Δgna-1 Δgna-3 mutants, which was partially suppressed by peptone supplementation. Stimulation of adenylyl cyclase is completely deficient in Δgna-1 Δ gna-2 Δgna-3 and Δgna-1 Δ gna-3 strains. Δgna-3 and Δ gna-1 Δgna-3 aerial hyphae and conidiation defects were suppressed by mutation of a PKA regulatory subunit. ^

El discurso periodístico de Gastón Talamón en torno al nacionalismo musical argentino : los escritos publicados en la revista ‘Tárrega’

Se explican los mecanismos receptivos que contribuyeron a mantener el ideario nacionalista y los procesos de identifi cación cultural, a través de la música, mediante el análisis de dos escritos del musicógrafo y crítico Gastón Talamón, publicados en 1926 en la revista Tárrega. Figura principal entre los teóricos del denominado ‘nacionalismo musical argentino’, los artículos se titulan “Tres nuevas nacionalidades musicales" y “El anti-argentinismo". La metodología utilizada apela a herramientas del análisis discursivo y el marco teórico entrecruza nociones de teoría de la recepción y teoría de la mediación, en línea con una historia socio-cultural de la música argentina.

Estudio genético y molecular de la respuesta a la vernalización en zanahoria (Daucus carota L.)

La zanahoria es una planta bienal de estación fría que requiere de un período de vernalización para florecer. Sin embargo, algunos cultivares adaptados a zonas más cálidas requieren de menor vernalización y son clasificados como anuales o de floración temprana. El objetivo de esta tesis fue determinar la base genética e identificar los genes y/o regiones cromosómicas involucradas en los requerimientos de vernalización en la zanahoria. Para ello fueron evaluadas a campo familias segregantes F1, F2, F3, RC1 y RC2 obtenidas a partir de un cruzamiento entre una planta anual y una bienal. En base a los patrones de segregación observados se concluyó que la anualidad, o bajos requerimientos de vernalización, estaría determinada por un gen simple dominante. Al evaluar introducciones de zanahoria anuales y bienales de diversos orígenes geográficos y sus cruzamientos, se volvió a observar la total dominancia de la anualidad y se encontró variabilidad en el ciclo entre materiales anuales y entre materiales bienales. Utilizando un método molecular que se basa en similitud completa (BLAST), no se encontró en el genoma de la zanahoria secuencias homólogas al gen FLC, el cual juega un rol central en la respuesta a la vernalización en Arabidopsis y otras especies como las Brassicas. Mediante la técnica de mapeo se encontró una región cromosómica ligada a la respuesta a la vernalización en zanahoria. La misma se localizó en un grupo de ligamiento con 78 marcadores moleculares a una distancia de 0,69 cM y de 0,79 cM de los marcadores más cercanos. Este mapa servirá como base para el desarrollo de marcadores moleculares ligados al carácter y en un futuro para el mapeo físico y secuenciación de la región de interés utilizando una librería génica de BACs de zanahoria.

El significado aritmológico de la Tríada y sus Proyecciones Filosófico-Religiosas

Bertrand Russell dio una solución lógica general a la definición de los números caracterizando al número tres como la clase que es correspondiente a todas las clases biunívocas con los grupos de tres. Para Frege la definición del número era una de las cruces de la comprensión de la aritmética. Hegel, por su parte, bajo el impulso de la triplicidad kantiana de los juicios sintéticos, teoréticos y prácticos y la influencia de la concepción trinitaria cristiana advertía que la contraposición de los opuestos al no ser contradictoria permitía el desarrollo deviniente. La aritmología pitagórica sobre los números naturales destacó la definición del tres caracterizada por su naturaleza de mediedad. Retomando estas bases de pura inteligibilidad y las especulaciones gnósticas sobre la tríada mostraremos sus proyecciones filosófico-religiosas.

La tensión entre las acciones reivindicativas y la proyección política de los sectores populares : Un estudio sobre los límites y las posibilidades de politización de las experiencias recientes de organización popular. El caso del Movimiento de Trabajadores Desocupados de Lanús

El presente plan se propone indagar sobre los procesos de politización de las prácticas y los vínculos sociales de los sectores populares organizados en torno a colectivos de trabajadores desocupados, a partir de las transformaciones políticas y sociales acontecidas en los últimos años. Buena parte de las expectativas y demandas de cambio social suscitadas recientemente se habían depositado en las experiencias de los movimientos piqueteros, en tanto agentes capaces de encarnar prácticas renovadas de organización política y alternativas colectivas de cambio social. Desde su irrupción pública, sin embargo, la literatura académica había advertido sobre las contradicciones que atravesaban a este movimiento social. Una de las dificultades más importantes radicaba en los inconvenientes encontrados por las organizaciones de desocupados para articular políticamente las demandas de subsistencia de sus bases sociales. En el marco de esta propuesta, se interpreta que estas dificultades dan cuenta de una tensión constitutiva que atraviesa al conjunto del movimiento. Esta tensión se expresa en el vínculo problemático entre las dimensiones reivindicativas y las dimensiones políticas de las experiencias piqueteras. En tal sentido, se pretende realizar un balance y una actualización crítica de las alternativas emergentes de construcción política popular, desde la trayectoria de una de las organizaciones de desocupados señaladas como caso ejemplar de renovación política y social. Se tratará, por tanto, de abordar las respuestas articuladas por esta organización a la tensión entre las cuestiones reivindicativas y las cuestiones políticas, en el contexto de las mutaciones producidas recientemente en el escenario político, especialmente a partir de la asunción del gobierno de Néstor Kirchner en el año 2003.

Empresarialidad y desarrollo local en la región rural periurbana de La Plata

El artículo se centra en la reflexión analítica de los participantes de un programa de desarrollo emprendedor en la Región Rural Periurbana centrada en el municipio de La Plata. El programa partió de una acción de extensión de la Universidad Nacional de La Plata (Olmos Emprende), cuyo desarrollo derivó en la articulación con programas más abarcativos de orden nacional y provincial. El primer objetivo es describir las condiciones específicas en las que se desarrolla la empresarialidad en América Latina y contextualizar la experiencia presentada. Posteriormente se hace una consideración del contexto territorial local que habría facilitado la participación en el programa teniendo en cuenta dos ámbitos regionales: por un lado la Región Rural Periurbana en general y por el otro lado, una serie de dimensiones locales que, además de factores tecno-económicos y marcos regulatorios estatales, incluyera circunstancias sociales y culturales. La pregunta que atraviesa el texto, focaliza la ubicación de la empresarialidad en el territorio, enfocado en sus posibilidades de desarrollo local. Observamos entonces las situaciones de interfase entre diversos actores territoriales, entre grupos de ciudadanos, diversas agencias estatales y la participación asimismo diversa de los agentes privados

Empresarialidad y desarrollo local en la región rural periurbana de La Plata

El artículo se centra en la reflexión analítica de los participantes de un programa de desarrollo emprendedor en la Región Rural Periurbana centrada en el municipio de La Plata. El programa partió de una acción de extensión de la Universidad Nacional de La Plata (Olmos Emprende), cuyo desarrollo derivó en la articulación con programas más abarcativos de orden nacional y provincial. El primer objetivo es describir las condiciones específicas en las que se desarrolla la empresarialidad en América Latina y contextualizar la experiencia presentada. Posteriormente se hace una consideración del contexto territorial local que habría facilitado la participación en el programa teniendo en cuenta dos ámbitos regionales: por un lado la Región Rural Periurbana en general y por el otro lado, una serie de dimensiones locales que, además de factores tecno-económicos y marcos regulatorios estatales, incluyera circunstancias sociales y culturales. La pregunta que atraviesa el texto, focaliza la ubicación de la empresarialidad en el territorio, enfocado en sus posibilidades de desarrollo local. Observamos entonces las situaciones de interfase entre diversos actores territoriales, entre grupos de ciudadanos, diversas agencias estatales y la participación asimismo diversa de los agentes privados

La tensión entre las acciones reivindicativas y la proyección política de los sectores populares : Un estudio sobre los límites y las posibilidades de politización de las experiencias recientes de organización popular. El caso del Movimiento de Trabajadores Desocupados de Lanús

El presente plan se propone indagar sobre los procesos de politización de las prácticas y los vínculos sociales de los sectores populares organizados en torno a colectivos de trabajadores desocupados, a partir de las transformaciones políticas y sociales acontecidas en los últimos años. Buena parte de las expectativas y demandas de cambio social suscitadas recientemente se habían depositado en las experiencias de los movimientos piqueteros, en tanto agentes capaces de encarnar prácticas renovadas de organización política y alternativas colectivas de cambio social. Desde su irrupción pública, sin embargo, la literatura académica había advertido sobre las contradicciones que atravesaban a este movimiento social. Una de las dificultades más importantes radicaba en los inconvenientes encontrados por las organizaciones de desocupados para articular políticamente las demandas de subsistencia de sus bases sociales. En el marco de esta propuesta, se interpreta que estas dificultades dan cuenta de una tensión constitutiva que atraviesa al conjunto del movimiento. Esta tensión se expresa en el vínculo problemático entre las dimensiones reivindicativas y las dimensiones políticas de las experiencias piqueteras. En tal sentido, se pretende realizar un balance y una actualización crítica de las alternativas emergentes de construcción política popular, desde la trayectoria de una de las organizaciones de desocupados señaladas como caso ejemplar de renovación política y social. Se tratará, por tanto, de abordar las respuestas articuladas por esta organización a la tensión entre las cuestiones reivindicativas y las cuestiones políticas, en el contexto de las mutaciones producidas recientemente en el escenario político, especialmente a partir de la asunción del gobierno de Néstor Kirchner en el año 2003.