Collodial

Targeted insertion of a plasmid by homologous recombination was demonstrated in zebrafish ES cell cultures. Two selection strategies were used to isolate ES cell colonies that contained targeted plasmid insertions in either the no tail or myostatin I gene. One selection strategy involved the manual isolation of targeted cell colonies that were identified by the loss of fluorescent protein gene expression. A second strategy used the diphtheria toxin A-chain gene in a positive-negative selection approach. Homologous recombination was confirmed by PCR, sequence and Southern blot analysis and colonies isolated using both selection methods were expanded and maintained for multiple passages...

OBJECTIVE: This paper reviewed the feasibility and accuracy of sentinel lymph node status in women with breast cancer. DESIGN: Clinical retrospective study. SETTING: Dept. of Obstetrics and Gynecology, 2nd Medical School, Teaching Hospital Motol, Praha. METHODS: Our trial involved 169 patients with breast cancer in the T1 and T2 stage without suspicion for axillary lymph node involvement. Lymphatic mapping was performed by sub dermal or subareolar injection of 99mTc labeled collodial human albumin (Senti-Scint) in the dose of 15 MBg one day before surgery...

In the current study, the first non-mammalian growth/differentiation factor (GDF) 11-like homolog was cloned from zebrafish. At the nucleotide level, zebrafish GDF11 is most similar to human GDF11 (79%), while the peptide is most similar to mouse GDF11 (78%). Phylogenetic analysis showed that the zebrafish GDF11 clusters with mammalian GDF11s. This study also cloned a second MSTN form in zebrafish most similar to Salmonid MSTN2 forms. Based on real time PCR, GDF11 is expressed in multiple adult tissues, with levels highest in whole heads and gonads, and expression is less ubiquitous when compared to MSTN expression...

Fibronectin (FN) is a highly conserved extracellular matrix protein that plays crucial roles in vertebrate embryogenesis. Previously, it was reported that zebrafish possess a single FN gene (fn1a). Here we report the presence of a second zebrafish FN gene (fn1b) that encodes a protein with a predicted primary structure that is similar to FNs identified in other vertebrates possessing 12 type I, 2 type II and 17 type III repeats including two alternative splice sites (EIIIA and EIIIB) and a variable region (V)...

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Although the zebrafish possesses several favourable characteristics that make it an ideal model for genetic studies of vertebrate development, one disadvantage of this model system is the absence of methods for the production of gene knockouts. The authors' laboratory, and others, are working to develop zebrafish pluripotent embryonic stem (ES) and primordial germ cell (PGC) cultures that can be used for cell-mediated gene transfer and the production of knockout mutant lines of fish. Progress has been made in developing short-term cell cultures that possess the ability to contribute to multiple tissues, including the germ line of a host embryo, and transgenic lines of zebrafish have been established using the embryo cell cultures...

The zebrafish is an established model for the genetic analysis of vertebrate development. Forward-genetic screens have generated thousands of mutations, and antisense-based methods have been used to transiently knockdown gene expression during embryogenesis. Although these methods have made the zebrafish a valuable system for the identification and functional characterization of developmentally important genes, one deficiency of the zebrafish model is the absence of methods to introduce targeted mutations to generate knockout lines of fish...

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Fibronectin is a highly conserved and well-characterized extracellular matrix protein that plays a crucial role in many cellular processes. In amphibians, birds and mammals, multiple isoforms of fibronectin are generated by differential RNA splicing at three exons. Previously, we identified a novel form of fibronectin (FN2) in zebrafish that possesses a truncated structure and is generated by a unique RNA splicing pattern. Unlike other fibronectins, FN2 exists as a monomer and localizes on the cell surface...

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As a first step toward developing a zebrafish model for investigating the role of sulfation in counteracting environmental estrogenic chemicals, we have embarked on the identification and characterization of cytosolic sulfotransferases (STs) in zebrafish. By searching the zebrafish expressed sequence tag database, we have identified two cDNA clones encoding putative cytosolic STs. These two zebrafish ST cDNAs were isolated and subjected to nucleotide sequencing. Sequence data revealed that the two zebrafish STs are highly homologous, being approximately 82% identical in their amino acid sequences...

Institutions are currently seeking alternative ways to deliver a full-line of course materials without acquiring additional staffing. Hence, faculty is charged with creating alternative ways to deliver or offer course content to students. The purpose of this study was to evaluate undergraduate and graduate performance and perception of teleconferencing versus traditional blackboard lectures. In the undergraduate course, we discovered that students performed equally as well on exams and provided favorable reviews of the course; however, the acceptance of this new format is lacking given the enrollment and number of students dropping, e...