Amphibian transcription factor IIIA proteins contain a sequence element functionally equivalent to the nuclear export signal of human immunodeficiency virus type 1 Rev.

The human immunodeficiency virus type 1 (HIV-1) Rev protein is required for nuclear export of late HIV-1 mRNAs. This function is dependent on the mutationally defined Rev activation domain, which also forms a potent nuclear export signal. Transcription factor IIIA (TFIIIA) binds to 5S rRNA transcripts and this interaction has been proposed to play a role in the efficient nuclear export of 5S rRNA in amphibian oocytes. Here it is reported that amphibian TFIIIA proteins contain a sequence element with homology to the Rev activation domain that effectively substitutes for this domain in inducing the nuclear export of late HIV-1 mRNAs. It is further demonstrated that this TFIIIA sequence element functions as a protein nuclear export signal in both human cells and frog oocytes. Thus, this shared protein motif may play an analogous role in mediating the nuclear export of both late HIV-1 RNAs and 5S rRNA transcripts.

Role of glutathione in the export of compounds from cells by the multidrug-resistance-associated protein.

Multidrug-resistance-associated protein (MRP) is a plasma membrane glycoprotein that can confer multidrug resistance (MDR) by lowering intracellular drug concentration. Here we demonstrate that depletion of intracellular glutathione by DL-buthionine (S,R)-sulfoximine results in a complete reversal of resistance to doxorubicin, daunorubicin, vincristine, and VP-16 in lung carcinoma cells transfected with a MRP cDNA expression vector. Glutathione depletion had less effect on MDR in cells transfected with MDR1 cDNA encoding P-glycoprotein and did not increase the passive uptake of daunorubicin by cells, indicating that the decrease of MRP-mediated MDR was not due to nonspecific membrane damage. Glutathione depletion resulted in a decreased efflux of daunorubicin from MRP-transfected cells, but not from MDR1-transfected cells, suggesting that glutathione is specifically required for the export of drugs from cells by MRP. We also show that MRP increases the export of glutathione from the cell and this increased export is further elevated in the presence of arsenite. Our results support the hypothesis that MRP functions as a glutathione S-conjugate carrier.

Enteropathogenic Escherichia coli contains a putative type III secretion system necessary for the export of proteins involved in attaching and effacing lesion formation.

Enteropathogenic Escherichia coli (EPEC) causes a characteristic histopathology in intestinal epithelial cells called the attaching and effacing lesion. Although the histopathological lesion is well described the bacterial factors responsible for it are poorly characterized. We have identified four EPEC chromosomal genes whose predicted protein sequences are similar to components of a recently described secretory pathway (type III) responsible for exporting proteins lacking a typical signal sequence. We have designated the genes sepA, sepB, sepC, and sepD (sep, for secretion of E. coli proteins). The predicted Sep polypeptides are similar to the Lcr (low calcium response) and Ysc (yersinia secretion) proteins of Yersinia species and the Mxi (membrane expression of invasion plasmid antigens) and Spa (surface presentation of antigens) regions of Shigella flexneri. Culture supernatants of EPEC strain E2348/69 contain several polypeptides ranging in size from 110 kDa to 19 kDa. Proteins of comparable size were recognized by human convalescent serum from a volunteer experimentally infected with strain E2348/69. A sepB mutant of EPEC secreted only the 110-kDa polypeptide and was defective in the formation of attaching and effacing lesions and protein-tyrosine phosphorylation in tissue culture cells. These phenotypes were restored upon complementation with a plasmid carrying an intact sepB gene. These data suggest that the EPEC Sep proteins are components of a type III secretory apparatus necessary for the export of virulence determinants.

Adenoviral E1B-55kDa protein inhibits yeast mRNA export and perturbs nuclear structure.

The mechanisms of export of RNA from the nucleus are poorly understood; however, several viral proteins modulate nucleocytoplasmic transport of mRNA. Among these are the adenoviral proteins E1B-55kDa and E4-34kDa. Late in infection, these proteins inhibit export of host transcripts and promote export of viral mRNA. To investigate the mechanism by which these proteins act, we have expressed them in Saccharomyces cerevisiae. Overexpression of either or both proteins has no obvious effect on cell growth. By contrast, overexpression of E1B-55kDa bearing a nuclear localization signal (NLS) dramatically inhibits cell growth. In this situation, the NLS-E1B-55kDa protein is localized to the nuclear periphery, fibrous material is seen in the nucleoplasm, and poly(A)+ RNA accumulates in the nucleus. Simultaneous overexpression of E4-34kDa bearing or lacking an NLS does not modify these effects. We discuss the mechanisms of selective mRNA transport.

Protein export from the nucleus requires the GTPase Ran and GTP hydrolysis.

Nuclei of digitonin-permeabilized cells that had been preloaded with a model transport substrate in a cytosol-dependent import reaction were subsequently incubated to investigate which conditions would result in export of transport substrate. We found that up to 80% of the imported substrate was exported when recombinant human Ran and GTP were present in the export reaction. Ran-mediated export was inhibited by nonhydrolyzable GTP analogs and also by wheat germ agglutinin but was unaffected by a nonhydrolyzable ATP analog. Moreover, a recombinant human Ran mutant that was deficient in its GTPase activity inhibited export. These data indicate that export of proteins from the nucleus requires Ran and GTP hydrolysis but not ATP hydrolysis. We also found that digitonin-permeabilized cells were depleted of their endogenous nuclear Ran, thus allowing detection of Ran as a limiting factor for export. In contrast, most endogenous karyopherin alpha was retained in nuclei of digitonin-permeabilized cells. Unexpectedly, exogenously added, fluorescently labeled Ran, although it accessed the nuclear interior, was found to dock at the nuclear rim in a punctate pattern, suggesting the existence of Ran-binding sites at the nuclear pore complex.

Cell cycle-regulated nuclear import and export of Cdc47, a protein essential for initiation of DNA replication in budding yeast.

The CDC47 gene was isolated by complementation of a cdc47 temperature-sensitive mutant in Saccharomyces cerevisiae and was shown to encode a predicted polypeptide, Cdc47, of 845 aa. Cdc47 belongs to the Cdc46/Mcm family of proteins, previously shown to be essential for initiation of DNA replication. Using indirect immunofluorescence microscopy and subcellular fractionation techniques, we show that Cdc47 undergoes cell cycle-regulated changes in its subcellular localization. At mitosis, Cdc47 enters the nucleus, where it remains until soon after the initiation of DNA replication, when it is rapidly exported back into the cytoplasm. Cdc47 protein levels do not vary with the cell cycle, but expression of CDC47 and nascent synthesis of Cdc47 occur late in the cell cycle, coinciding with mitosis. Together, these results show that Cdc47 is not only imported into the nucleus at the end of mitosis but is also exported back into the cytoplasm at the beginning of S phase. The observation that Cdc47 is exported from the nucleus at the beginning of S phase has important implications for how initiation of DNA replication is controlled.

Comparison between clinical results of two diffractive multifocal lenses with the same platform but different additions

PURPOSE: To evaluate visual results with two multifocal diffractive lenses designed with the same platform but with different additions. SETTING: Grupo Innova Ocular clinics. METHODS: A total of 50 eyes from 50 patients were included. Group 1 (n = 25) was implanted with the TECNIS® 1 ZLB +3.25 and group 2 (n = 25) with the TECNIS® 1 ZKB +2.75. Patients were assessed at 24 hours, 1 week and 1 month postoperatively. At surgical discharge, corrected (CDVA) and uncorrected distance visual acuity (UCDVA), near visual acuity (VA) at 25, 40 and 80 cm, visual quality and the defocus curve were measured. RESULTS: Changes in sphere and spherical equivalent were statistically significant (p<0.01) in both groups at 1 week and 1 month compared to preoperative values. In group 1, UCDVA logMAR at 1 month was 0.06 ± 0.02. In group 2, UCDVA at 1 month was 0.03 ± 0.03. In near vision, the TECNIS® 1 ZLB group obtained a VA logMAR of 0.35 ± 0.02 at 25 cm, 0.13 ± 0.02 at 40 cm and 0.27 ± 0.02 at 80 cm, while in the TECNIS® 1 ZKB group, the values were 0.38  ± 0.03, 0.14 ± 0.03 and 0.23 ± 0.06, respectively. No statistically significant differences were found either when results for visual quality were compared. CONCLUSION: Both the TECNIS® 1 ZLB and TECNIS® 1 ZKB are excellent options for obtaining good distance and near vision, in addition to providing good intermediate vision, especially at distances such as those required for working with computers.

An experience on mechatronics teaching on undergraduate students by means of the Skybot platform: from classroom to robot competition

This paper analyzes the learning experiences and opinions from a group of undergraduate students in a course about Robotics. The contents of this course were taught as a set of seminars. In each seminar, the student learned interdisciplinary knowledge of computer science, control engineering, electronics and other fields related to Robotics. The aim of this course is that the students are able to design and implement their own and custom robotic solution for a series of tests planned by the teachers. These tests measure the behavior and mechatronic features of the students' robots. Finally, the students' robots are confronted with some competitions. In this paper, the low-cost robotic architecture used by the students, the contents of the course, the tests to compare the solutions of students and the opinion of them are amply discussed.

A viability analysis of a secure VoIP and instant messaging system on a Pocket PC platform

We propose a secure full-duplex VoIP and instant messaging system on a Pocket PC platform, allowing for session key transport using a public-key protocol and encrypted text or voice communication using a private-key algorithm. The full-duplex VoIP scheme presents good performance for long duration communication over LAN networks.

Java software platform for the development of advanced robotic virtual laboratories

This article presents an interactive Java software platform which enables any user to easily create advanced virtual laboratories (VLs) for Robotics. This novel tool provides both support for developing applications with full 3D interactive graphical interface and a complete functional framework for modelling and simulation of arbitrary serial-link manipulators. In addition, its software architecture contains a high number of functionalities included as high-level tools, with the advantage of allowing any user to easily develop complex interactive robotic simulations with a minimum of programming. In order to show the features of the platform, the article describes, step-by-step, the implementation methodology of a complete VL for Robotics education using the presented approach. Finally, some educational results about the experience of implementing this approach are reported.

Integrated taphonomy in an open-marine platform: The Lower Cretaceous of Sierra Helada (Betic Cordillera, SE Spain)

A detailed taphonomic analysis was carried out on the lower Albian deposits of the Sierra Helada section (Alicante, Betic Cordillera, southeastern Spain). Ten taphonomic characters were studied and ten skeletal concentrations were defined on the basis of taphonomic features and the dominant taxa. Cluster analysis was performed on the dataset represented by the abundance of the taphonomic characters in each skeletal concentration. This enabled the definition of four different taphonomic categories: 1) skeletal concentrations characterized by the presence of fossils preserved in life position, 2) skeletal concentrations showing very little physical reworking, 3) skeletal concentrations related to high-energy background conditions, and 4) skeletal concentrations produced by medium- to high-energy events. Four taphofacies were defined on the basis of the main sedimentological features and the most representative skeletal concentrations. Taphofacies A represents the low energy outer platform, rich in skeletal concentrations with echinoids in life position and only slightly reworked. The second taphofacies (taphofacies B) is very rich in reworked echinoid tests and calcarenitic beds and records the transition to shallower areas, while taphofacies C shows abundant thick-bedded calcarenites and skeletal concentrations produced by sediment transport and rapid deposition. Finally, cross-bedded grainstone beds, which are rich in fine-grained fragmented, locally reoriented bioclasts (taphofacies D), record the existence of shifting sandy dunes in the shallow inner part of the platform.

Evaluating Taphonomic Bias in a Storm-Disturbed Carbonate Platform: Effects of Compositional and Environmental Factors in Lower Jurassic Brachiopod Accumulations (Eastern Subbetic Basin, Spain)

In order to evaluate taxonomic and environmental control on the preservation pattern of brachiopod accumulations, sedimentologic and taphonomic data have been integrated with those inferred from the structure of brachiopod accumulations from the easternmost Lower Jurassic Subbetic deposits in Spain. Two brachiopod communities (Praesphaeroidothyris and Securina communities) were distinguished showing a mainly free-lying way of life in soft-bottom habitats. Three taphofacies are discriminated based on proportion of disarticulation, fragmentation, packing, and shell filling. Taphofacies 1 is represented by thinly fragmented, dispersed brachiopod shells in wackestone beds. Taphofacies 2 is spatially restricted to small lenses where shells are poorly fragmented, rarely disarticulated, usually void filled, and highly packed. Taphofacies 3 is represented by mud or cement filled, loosely packed, articulated brachiopods forming large pocket-like structures. Temporal and spatial averaging were minimally involved in taphofacies 2 and 3. It is interpreted that patchy preservation implies preservation of primary original patchiness of brachiopod communities on the seafloor. The origin of shell-rich taphofacies (2 and 3) is related to rapid burial due to episodic storm activity, while shell-poor taphofacies 1 records background conditions. The nature and comparative diversity of these taphofacies underscores the importance of rapid burial for shell beds preservation. Differences in preservation between taphofacies 2 and 3 are mainly related to environmental criteria, most importantly storm energy and water depth. In contrast, the taxonomic-specific pattern of the communities is a subordinate element of control, controlling only minor within-taphofacies differences in preservation.