Telomere-based proliferative lifespan barriers in Werner-syndrome fibroblasts involve both p53-dependent and p53-independent mechanisms

Werner-syndrome fibroblasts have a reduced in vitro life span before entering replicative senescence. Although this has been thought to be causal in the accelerated ageing of this disease, controversy remains as to whether Werner syndrome is showing the acceleration of a normal cellular ageing mechanism or the occurrence of a novel Werner-syndrome-specific process. Here, we analyse the signalling pathways responsible for senescence in Werner-syndrome fibroblasts. Cultured Werner-syndrome (AG05229) fibroblasts senesced after approximately 20 population doublings with most of the cells having a 2N content of DNA. This was associated with hypophosphorylated pRb and high levels of p16(Ink4a) and p21(Waf1). Senescent AG05229 cells re-entered the cell cycle following microinjection of a p53-neutralizing antibody. Similarly, production of the human papilloma virus 16 E6 oncoprotein in presenescent AG05229 cells resulted in senescence being bypassed and extended cellular life span. Werner-syndrome fibroblasts expressing E6 did not proliferate indefinitely but reached a second proliferative lifespan barrier, termed M(int), that could be bypassed by forced production of telomerase in post-M1 E6-producing cells. The conclusions from these studies are that: (1) replicative senescence in Werner-syndrome fibroblasts is a telomere-induced p53-dependent event; and (2) the intermediate lifespan barrier M(int) is also a telomere-induced event, although it appears to be independent of p53. Werner-syndrome fibroblasts resemble normal human fibroblasts for both these proliferative lifespan barriers, with the strong similarity between the signalling pathway linking telomeres to cell-cycle arrest in Werner-syndrome and normal fibroblasts providing further support for the defect in Werner syndrome causing the acceleration of a normal ageing mechanism

Paleobiology of titanosaurs: reproduction, development, histology, pneumaticity, locomotion and neuroanatomy from the South American fossil record

Fil: García, Rodolfo A.. Instituto de Investigación en Paleobiología y Geología. Museo Provincial Carlos Ameghino. Cipolletti; ArgentinaFil: Salgado, Leonardo. Instituto de Investigación en Paleobiología y Geología. General Roca. Río Negro; ArgentinaFil: Fernández, Mariela. Inibioma-Centro Regional Universitario Bariloche. Bariloche. Río Negro; ArgentinaFil: Cerda, Ignacio A.. Instituto de Investigación en Paleobiología y Geología. Museo Provincial Carlos Ameghino. Cipolletti; ArgentinaFil: Carabajal, Ariana Paulina. Museo Carmen Funes. Plaza Huincul. Neuquén; ArgentinaFil: Otero, Alejandro. Museo de La Plata. Universidad Nacional de La Plata; ArgentinaFil: Coria, Rodolfo A.. Instituto de Paleobiología y Geología. Universidad Nacional de Río Negro. Neuquén; ArgentinaFil: Fiorelli, Lucas E.. Centro Regional de Investigaciones Científicas y Transferencia Tecnológica. Anillaco. La Rioja; Argentin

Evasion of p53-mediated growth control occurs by three alternative mechanisms in transformed thyroid epithelial cells

Using the thyroid as a model of multistep epithelial tumorigenesis, we have used representative cell lines to correlate the degree of malignant transformation with the functional status of p53 and the integrity of cell-cycle check-points. Three distinct phenotypes were observed: Type I lines, derived from poorly-differentiated human thyroid cancers, expressed high levels of mutant p53 protein; Type II, also poorly-differentiated but derived from rat, showed over-expression of wild-type (wt) p53 with marked cell-cell heterogeneity: Type III, from well-differentiated human cancers, contained uniformly low levels of wt p53. All cell lines containing wt p53 retained a near-normal induction of p53 by DNA damage. However, the ability to undergo growth arrest differed strikingly. Whereas Type I and II lines had lost both G2/M and G1/S check points, Type III cells retained both. In Type III cells, as in diploid human fibroblasts, mutant p53 expression specifically abrogated G1/S check-point function with no other change in phenotype. These data demonstrate 3 mechanisms for evasion of p53 growth control: (i) direct mutation (ii) indirect inactivation, or (iii) 'avoidance' of activation, most probably due to failure to reach a critical threshold of DNA damage

Paleobiología de Titanosaurios de Sudamérica: reproducción, desarrollo, histología, neumaticidad, locomoción y neuroanatomía

Much of the current paleobiological knowledge on titanosaur sauropods was attained in just the last fifteen years, in particular that related to reproductive and developmental biology. Recent years have also seen progress on other poorly explored topics, such as pneumaticity, muscle architecture and locomotion, and endocast reconstruction and associated structures. Some titanosaurs laid numerous, relatively small Megaloolithidae eggs (with diameters ranging from 12 to 14 cm) in nests dug In the ground and, as known from the South American records, probably eggs of the multispherulitic morphotype. During ontogeny, certain titanosaurs displayed some variations in cranial morphology, some of them likely associated with the differing feeding habits between hatchlings and adults. The bone tissue of some adult titanosaurs was rapidly and cyclically deposited and shows a greater degree of remodeling than in other sauropods. Saltasaurines in particular show evidence of postcranial skeletal pneumaticity in both axial and appendicular skeleton, providing clues about soft tissue anatomy and the structure of the respiratory system. Titanosaurs, like all sauropods, were characterized by being fully quadrupedal, although some appendicular features and putative trackways indicate that their stance was not as columnar as in other sauropods. These anatomical peculiarities are significantly developed In saltasaurines, a derived group of titanosaurs. Compared with other sauropods, some titanosaurs seem to have had very poor olfaction but would have been capable of capturing sounds In a relatively wide range of high frequencies, although not to the extent of living birds.El conocimiento paleobiológico de los saurópodos titanosaurios, particularmente su reproducción y biología del desarrollo, fue alcanzado recién en los últimos quince años. En estos últimos años también se ha avanzado en temas poco explorados hasta el momento, como la neumatización, su arquitectura muscular y locomoción y la reconstrucción de partes blandas como el cerebro y estructuras asociadas. Algunos titanosaurios depositan sus numerosos y pequeños huevos megaloolitidos en nidos excavados sobre el suelo. Durante la ontogenia ciertos titanosaurios exponen algunas variaciones en su morfología craneana, algunas de estas probablemente asociadas con las diferentes maneras de alimentarse que tendrían los juveniles y los adultos. El tejido óseo de algunos titanosaurios adultos se habría depositado rápido y cíclicamente, exponiendo una mayor remodelación que en otros saurópodos. Los titanosaurios, particularmente los saltasaurinos, exponen una neumaticidad postcraneal en el esqueleto axial y apendicular, este carácter permite Inferir la anatomía de sus tejidos blandos y de su sistema respiratorio. Los titanosaurios, como todos los saurópodos, estaban caracterizados por ser cuadrúpedos, aunque algunos caracteres apendiculares y las huellas indican que su postura no habría sido tan columnar como en otros saurópodos. Aquellas peculiaridades anatómicas están notoriamente desarrolladas en los saltasaurinos, un grupo de titanosaurios derivados. Comparado con otros saurópodos, algunos titanosaurios parecen haber tenido un pobre sentido del olfato, sin embargo estos habrían tenido la capacidad de captar sonidos de alta frecuencia en un rango relativamente amplio, aunque no tanto como las aves actuales.Fil: Garcia, Rodolfo Andres. Provincia de Río Negro. Museo Provincial “Carlos Ameghino”. Instituto de Investigación en Paleobiología y Geología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Salgado, Leonardo. Provincia de Río Negro. Instituto de Investigación en Paleobiología y Geología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Fernández, Mariela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Patagonia Norte. Instituto de Investigación en Biodiversidad y Medioambiente; Argentina. Universidad Nacional del Comahue. Centro Regional Universitario Bariloche; ArgentinaFil: Cerda, Ignacio Alejandro. Provincia de Río Negro. Museo Provincial “Carlos Ameghino”. Instituto de Investigación en Paleobiología y Geología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Paulina Carabajal, Ariana. Provincia del Neuquen. Municipalidad de Plaza Huincul. Museo "Carmen Funes"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Otero, Alejandro. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Departamento Científico de Paleontología de Vertebrados; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Coria, Rodolfo Anibal. Universidad Nacional de Rio Negro. Sede Alto Valle. Instituto de Investigaciones en Paleobiologia y Geologia; Argentina. Provincia del Neuquen. Municipalidad de Plaza Huincul. Museo "Carmen Funes"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Fiorelli, Lucas Ernesto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Regional de Investigaciones Cientificas y Transferencia Tecnológica de Anillaco; Argentin