Activation and Functional Analysis of Janus Kinase 2 in BA/F3 Cells Using the Coumermycin/Gyrase B System

Janus kinase 2 (Jak2) protein tyrosine kinase plays an important role in interleukin-3– or granulocyte–macrophage colony-stimulating factor–mediated signal transduction pathways leading to cell proliferation, activation of early response genes, and inhibition of apoptosis. However, it is unclear whether Jak2 can activate these signaling pathways directly without the involvement of cytokine receptor phosphorylation. To investigate the specific role of Jak2 in the regulation of signal transduction pathways, we generated gyrase B (GyrB)–Jak2 fusion proteins, dimerized through the addition of coumermycin. Coumermycin induced autophosphorylation of GyrB–Jak2 fusion proteins, thus bypassing receptor activation. Using different types of chimeric Jak2 molecules, we observed that although the kinase domain of Jak2 is sufficient for autophosphorylation, the N-terminal regions are essential for the phosphorylation of Stat5 and for the induction of short-term cell proliferation. Moreover, coumermycin-induced activation of Jak2 can also lead to increased levels of c-myc and CIS mRNAs in BA/F3 cells stably expressing the Jak2 fusion protein with the intact N-terminal region. Conversely, activation of the chimeric Jak2 induced neither phosphorylation of Shc or SHP-2 nor activation of the c-fos promoter. Here, we showed that the GyrB–Jak2 system can serve as an excellent model to dissect signals of receptor-dependent and -independent events. We also obtained evidence indicating a role for the N-terminal region of Jak2 in downstream signaling events.

The Arabidopsis Information Resource (TAIR): a comprehensive database and web-based information retrieval, analysis, and visualization system for a model plant

Arabidopsis thaliana, a small annual plant belonging to the mustard family, is the subject of study by an estimated 7000 researchers around the world. In addition to the large body of genetic, physiological and biochemical data gathered for this plant, it will be the first higher plant genome to be completely sequenced, with completion expected at the end of the year 2000. The sequencing effort has been coordinated by an international collaboration, the Arabidopsis Genome Initiative (AGI). The rationale for intensive investigation of Arabidopsis is that it is an excellent model for higher plants. In order to maximize use of the knowledge gained about this plant, there is a need for a comprehensive database and information retrieval and analysis system that will provide user-friendly access to Arabidopsis information. This paper describes the initial steps we have taken toward realizing these goals in a project called The Arabidopsis Information Resource (TAIR) (www.arabidopsis.org).

Expression and analysis of presenilin 1 in a human neuronal system: localization in cell bodies and dendrites.

Mutations in the recently identified presenilin 1 gene on chromosome 14 cause early onset familial Alzheimer disease (FAD). Herein we describe the expression and analysis of the protein coded by presenilin 1 (PS1) in NT2N neurons, a human neuronal model system. PS1 was expressed using recombinant Semliki Forest virions and detected by introduced antigenic tags or antisera to PS1-derived peptides. Immunoprecipitation revealed two major PS1 bands of approximately 43 and 50 kDa, neither of which were N-glycosylated or O-glycosylated. Immunoreactive PS1 was detected in cell bodies and dendrites of NT2N neurons but not in axons or on the cell surface. PS1 was also detected in BHK cells, where it was also intracellular and colocalized with calnexin, a marker for the rough endoplasmic reticulum. A mutant form of PS1 linked to FAD did not differ from the wild-type protein at the light microscopic level. The model system described here will enable studies of the function of PS1 in human neurons and the role of mutant PS1 in FAD.

Lens complementation system for the genetic analysis of growth, differentiation, and apoptosis in vivo.

A genetic approach has been established that combines the advantages of blastocyst complementation with the experimental attributes of the developing lens for the functional analysis of genes governing cellular proliferation, terminal differentiation, and apoptosis. This lens complementation system (LCS) makes use of a mutant mouse strain, aphakia (ak), homozygotes of which fail to develop an ocular lens. We demonstrate that microinjection of wild-type embryonic stem (ES) cells into ak/ak blastocysts produces chimeras with normal ES-cell-derived lenses and that microinjection of Rb-/- ES cells generates an aberrant lens phenotype identical to that obtained through conventional gene targeting methodology. Our determination that a cell autonomous defect underlies the aphakia condition assures that lenses generated through LCS are necessarily ES-cell-derived. LCS provides for the rapid phenotypic analysis of loss-of-function mutations, circumvents the need for germ-line transmission of null alleles, and, most significantly, facilitates the study of essential genes whose inactivation is associated with early lethal phenotypes.

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.

Profitability analysis of a femtosecond laser system for cataract surgery using a fuzzy logic approach

AIM: To define the financial and management conditions required to introduce a femtosecond laser system for cataract surgery in a clinic using a fuzzy logic approach. METHODS: In the simulation performed in the current study, the costs associated to the acquisition and use of a commercially available femtosecond laser platform for cataract surgery (VICTUS, TECHNOLAS Perfect Vision GmbH, Bausch & Lomb, Munich, Germany) during a period of 5y were considered. A sensitivity analysis was performed considering such costs and the countable amortization of the system during this 5y period. Furthermore, a fuzzy logic analysis was used to obtain an estimation of the money income associated to each femtosecond laser-assisted cataract surgery (G). RESULTS: According to the sensitivity analysis, the femtosecond laser system under evaluation can be profitable if 1400 cataract surgeries are performed per year and if each surgery can be invoiced more than $500. In contrast, the fuzzy logic analysis confirmed that the patient had to pay more per surgery, between $661.8 and $667.4 per surgery, without considering the cost of the intraocular lens (IOL). CONCLUSION: A profitability of femtosecond laser systems for cataract surgery can be obtained after a detailed financial analysis, especially in those centers with large volumes of patients. The cost of the surgery for patients should be adapted to the real flow of patients with the ability of paying a reasonable range of cost.

System Analysis of Innovative Projects in Mechanical Engineering

Modern production and innovation processes in industrial corporations differ by complex nature of modern technologies and organizational solutions. Adequate methodological approaches to their analysis and evaluation of efficiency are required. The present study contains the conceptand the model of operational and innovation program for which formulated theoretical and methodological principles and systematic statement of the general problem of planning, designed to optimize the process of resource usage under different criteria, including: investment, financial, technological, industrial and information capabilities of the corporation.

Analysis of the reproductive system of female vampire squid

Analysis of the reproductive system of female vampire squid to determine reproductive strategy and fecundity of vampire squid, accessioned in the Santa Barbara Museum of Natural History and collected in 60, 70s from off southern California.

Analysis of H.R. 12889 : a bill to amend an act entitled "An act to establish a uniform system of bankruptcy throughout the United States" approved July 1, 1898, and acts amendatory thereof and supplementary thereto and to repeal section 76 thereof and all acts and parts of acts inconsistent therewith /

"Containing amendments proposed by the National Bankruptcy Conference."