Comparison of different protocols for the extraction of microbial DNA from reef corals

This study aimed to test different protocols for the extraction of microbial DNA from the coral Mussismilia harttii. Four different commercial kits were tested, three of them based on methods for DNA extraction from soil (FastDNA SPIN Kit for soil, MP Bio, PowerSoil DNA Isolation Kit, MoBio, and ZR Soil Microbe DNA Kit, Zymo Research) and one kit for DNA extraction from plants (UltraClean Plant DNA Isolation Kit, MoBio). Five polyps of the same colony of M. harttii were macerated and aliquots were submitted to DNA extraction by the different kits. After extraction, the DNA was quantified and PCR-DGGE was used to study the molecular fingerprint of Bacteria and Eukarya. Among the four kits tested, the ZR Soil Microbe DNA Kit was the most efficient with respect to the amount of DNA extracted, yielding about three times more DNA than the other kits. Also, we observed a higher number and intensities of DGGE bands for both Bacteria and Eukarya with the same kit. Considering these results, we suggested that the ZR Soil Microbe DNA Kit is the best adapted for the study of the microbial communities of corals.

Muerte celular programada en protozoarios: el caso de Giardia intestinalis / Programmed cell death in protozoa: Giardia intestinalis’s case

Giardia intestinalis es considerado uno de los eucariotas más antiguos y su poca complejidad representa una valiosa oportunidad para desentrañar los misterios de procesos vitales de eucariotas más complejos. Esta característica única de G. intestinalis y el hecho de que su genoma esté completamente secuenciado y disponible, y que todo su ciclo de vida puede ser reproducido in vitro, hacen de este parásito un modelo ideal para estudiar mecanismos celulares, entre ellos, la muerte celular programada. Desde el punto de vista morfológico y molecular, la apoptosis es uno de los tipos más complejos de muerte celular programada, la cual es un proceso normal durante el desarrollo celular, y tiene un papel esencial en el control de la proliferación celular y en la respuesta a retos inmunológicos o a daños celulares. Recientemente, se ha reportado que en protozoos, entre ellos Giardia, podría ocurrir un tipo de muerte celular programada similar a la apoptosis y los resultados de nuestros laboratorios apoyan esta hipótesis; sin embargo, no se han identificado hasta el momento las moléculas relacionadas con los procesos de apoptosis en estos parásitos. La presente revisión abarca una descripción de la morfología y estructura de las formas de vida de G. intestinalis, de su ciclo biológico, de la parasitosis que causa y de las estrategias quimioterapéuticas para su tratamiento. Asimismo, se hace un repaso de lo que hasta ahora se conoce sobre apoptosis en protozoarios, y específicamente en G. intestinalis, y se describen algunos resultados de nuestro grupo que apoyan la existencia de muerte celular programada en este parásito.

Giardia intestinalis is an early-branching eukaryote and its low complexity represents a valuable opportunity to unravel the mysteries of essential processes in more complex eukaryotes. In addition, the genome of G. intestinalis is completely sequenced and its entire life cycle can be reproduced in vitro. All these characteristics make of Giardia an ideal model for studying cellular mechanisms, such as programmed cell death. Apoptosis is one of the most complex types of programmed cell death and plays an essential role during cell development, cell proliferation and immune response. Recently it has been reported that in Giardia can take place events that resemble apoptosis and although our results support this hypothesis, molecules involved in this process have not yet been identified. This review includes a description of the morphology and structure of G. intestinalis, its life cycle, the disease that causes and the strategies for its treatment. In addition, we review what is known about apoptosis in protozoa, and specifically in G. intestinalis, and describe some results from our group supporting the existence of apoptosis-like programmed cell death in this parasite.

Protein synthesis in eukaryotes: The growing biological relevance of cap-independent translation initiation

Ribosome recruitment to eukaryotic mRNAs is generally thought to occur by a scanning mechanism, whereby the 40S ribosomal subunit binds in the vicinity of the 5'cap structure of the mRNA and scans until an AUG codon is encountered in an appropriate sequence context. Study of the picornaviruses allowed the characterization of an alternative mechanism of translation initiation. Picornaviruses can initiate translation via an internal ribosome entry segment (IRES), an RNA structure that directly recruits the 40S ribosomal subunits in a cap and 5' end independent fashion. Since its discovery, the notion of IRESs has extended to a number of different virus families and cellular RNAs. This review summarizes features of both cap-dependent and IRES-dependent mechanisms of translation initiation and discusses the role of cis-acting elements, which include the 5'cap, the 5'-untranslated region (UTR) and the poly(A) tail as well as the possible roles of IRESs as part of a cellular stress response mechanism and in the virus replication cycle.

Perfiles ribosomales en miometrio normal y patologico / Ribosomal profiles in normal and pathological myometrium

Se estudian los perfiles ribosomales en un miometrio normal en menopausia, y tres miometrios normales al final del embarazo. Se estudian tambien estos perfiles ribosomales en tres miometrios patologicos en adenomiosis, en miomatosis y en sarcomatosis. La mayor actividad en sintesis proteica miometrial fue revelada en orden decreciente por la adenomiosis, la sarcomatosis, la miomatosis, el embarazo a termino y finalmente la menopausia.