A recessive mutation leading to vertebral ankylosis in zebrafish is associated with amino acid alterations in the homologue of the human membrane-associated guanylate kinase DLG3.

We describe the characterization of the zebrafish homologue of the human gene DLG3. The zebrafish dlg3 gene encodes a membrane-associated guanylate kinase containing a single PDZ domain. This gene was cloned using a gene-trap construct inserted in the gene's first intron. The insertion co-segregates with a viable mutation called humpback (hmp), which leads to formation of ankylotic vertebrae in adult fishes. Insertion and mutation have both been mapped to chromosome 12, in a segment which is syntenic with region p12 to q12 of human chromosome 17. The hmp mutant phenotype, however, appears to be due to two point mutations in the guanylate kinase domain rather than to the transgene insertion itself. The results of this study are discussed in the light of the possible function of the guanylate kinase domain.     

Global introductions of salmon and trout in the genus <Emphasis Type="Italic">Oncorhynchus</Emphasis>: 1870–2007

The purpose of this review is to provide a global perspective on Oncorhynchus salmonine introductions and put-and-take fisheries based on modern stocking programs, with special emphasis on freshwater ecosystems. We survey the global introductions of nine selected salmonines of the genus Oncorhynchus: golden trout, cutthroat trout, pink salmon, chum salmon, coho salmon, masu/cherry salmon, rainbow trout/steelhead, sockeye salmon/kokanee, and chinook salmon. The information is organized on a geographical basis by continent, and then by species and chronology. Two different objectives and associated definitions of ‘success’ for introductions are distinguished: (a) seed introduction: release of individuals with the purpose of creating a wild-reproducing, self-sustaining population; and (b) put-and-take introduction: release of individuals with the purpose of maintaining some level of wild population abundance, regardless of wild reproduction. We identify four major phenomena regarding global salmonine introductions: (1) general inadequacy of documentation regarding introductions; (2) a fundamental disconnect between management actions and ecological consequences of introductions; (3) the importance of global climate change on success of previous and future introductions; and (4) the significance of aquaculture as a key uncertainty in accidental introductions. We conclude this review with a recognition of the need to terminate ongoing stocking programs for introduced salmonines worldwide.     

PHYLOGENETIC ANALYSIS OF THE MALE ONTOGENETIC PROGRAM IN AQUATIC AND TERRESTRIAL MONOCOTYLEDONS

The male program of ontogeny in flowering plants encompasses the events from meiosis of microsporocytes to fertilization. Three main sequences are discussed; the deposition of cell walls, changes in cytoplasmic organelles, and the program of nuclear divisions leading to the formation of two sperm cells and a vegetative cell in each pollen grain. Variations in these ontogenetic sequences are particularly apparent in the monocotyledons, which exhibit diversity in pollen morphology, wall structure, and mode of pollination. The male program of development has been compared in selected terrestrial monocotyledons belonging to the Liliaceae and Gramineae and aquatic members of the Cymodoceaceae, Najadaceae, and Zannichelliaceae. A total of 26 characters from the male program are discussed and then used to construct a cladogram derived only from developmental data for the five species. The polarity of only a few of the character transformations has been determined directly by observation of developmental sequences; most have been interpreted by outgroup analysis.     

Phosphoproteins in dictyostelium discoideum

The phosphoproteins of Dictyostelium discoideum were compared at different stages of development by polyacrylamide gel electrophoresis. Certain phosphoproteins of vegetative amoebae were conserved while others appeared and disappeared during development. Four major phosphoproteins with apparent subunit molecular weights of 50,000, 47,000, 38,000, and 34,000 disappeared precociously in response to exogenous cAMP. Two membranal phosphoproteins, with apparent subunit molecular weights of 80,000 and 81,000, appeared precociously in response to added cAMP. One of these phosphoproteins, molecular weight of 80,000, has been identified tentatively as the “contact site A” glycoprotein. Another membranal protein, with apparent subunit molecular weight of 42,000, unaffected in its appearance by cAMP, has been identified tentatively as phosphoactin.     

Spontaneous Aggregation and Cytotoxicity of the β-Amyloid Aβ1–40: A Kinetic Model

The time dependency of the spontaneous aggregation of the fibrillogenic β-Amyloid peptide, Aβ^1–40, was measured by turbidity, circular dichroism, HPLC, and fluorescence polarization. The results by all methods were comparable and they were most consistent with a kinetic model where the peptide first slowly forms an activated monomeric derivative (AM), which is the only species able to initiate, by tetramerization, the formation of linear aggregates. The anti-Aβ antibody 6E10, raised against residues 1–17, at concentrations of 200–300 nM delayed significantly the aggregation of 50 μM amyloid peptide. The anti–Aβ antibody 4G8, raised against residues 17–24, was much less active in that respect, while the antibody A162, raised against the C-terminal residues 39–43 of the full-length Aβ was totally inactive at those concentrations. Concomitant with the aggregation experiments, we also measured the time dependency of the Aβ^1–40–induced toxicity toward SH-EP1 cells and hippocampal neurons, evaluated by SYTOX Green fluorescence, lactate dehydrogenase release, and activation of caspases. The extent of cell damage measured by all methods reached a maximum at the same time and this maximum coincided with that of the concentration of AM. According to the kinetic scheme, the latter is the only transient peptide species whose concentration passes through a maximum. Thus, it appears that the toxic species of Aβ^1–40 is most likely the same transient activated monomer that is responsible for the nucleation of fibril formation. These conclusions should provide a structural basis for understanding the toxicity of Aβ^1–40 in vitro and possibly in vivo.     

Uteroferrin: A protein in search of a function

Uteroferrin, a purple-colored, iron-containing acid phosphatase, with many of the properties of a lysosomal hydrolase, transports iron from the mother to the conceptus in pregnant pigs. Uteroferrin, however, is but one member of what may be a broad class of iron-containing phosphatases with unusual spectral properties which result from a novel type of di-iron active site. The biological function of uteroferrin is unknown. We argue here that the in vivo function of uteroferrin, despite its undoubted ability to act as a potent acid phosphatase, is that of a transplacental iron transporter.     

Quantitative assay of deoxyribonuclease activity after isoelectric focusing in polyacrylamide gels: pH control and effects of enzyme diffusion

A method for the quantitative assay of nuclease activity in crude cell lysates after isoelectric focusing (IEF) in polyacrylamide slab gels is described. After IEF, an agarose overlay gel containing DNA is placed on the IEF gel and the nuclease activity quantified by the loss of ethidium bromide fluorescence of the DNA. With this method a linear response was obtained for 1 to 10 ng of DNase I. Various methods of pH equilibration after IEF were also evaluated. The use of a high buffer concentration in the overlay gel is recommended to control the pH during the enzyme reaction. An analytical solution for the diffusion of enzymes from the IEF gel to the overlay gel is also presented and an equation that may be used to choose optimum times for transfer of the enzyme from the IEF gel to the overlay gel is given.     

DISTANCE DATA REVISITED

Abstract— Objections to my earlier demonstration, that the branch lengths of trees fitted to distance matrices have no physical interpretation, are shown to be ill-founded. In particular the contention of Felsenstein, that fitted lengths estimate expectations of amounts of change, is shown to lead to a paradox. A method is introduced for constructing multiple trees of optimal or near-optimal fit to distance data, and this is found to give better performance than previous methods. Most published trees based on distances have been poorly chosen. Consensus trees of several trees with near-optimal fit are found to be quite poorly resolved, and it appears that molecular distances seldom provide much useful information on phylogenetic relationships.     

Metabolism of carbon tetrachloride to phosgene

Aerobic incubation of hepatic microsomal fractions in the presence of carbon tetrachloride, NADPH and cysteine resulted in the formation of phosgene which was identified by gas chromatography/mass spectrometry as the adduct, 2-oxothiazolidine-4-carboxylic acid, formed by its reaction with cysteine. [¹³C]-Carbon tetrachloride was metabolized to 2-[¹³C]-oxothiazolidine-4-carboxylic acid the , when carbon tetrachloride was incubated in the presence of [¹⁸O]-O₂, 2- [¹⁸O]-oxothiazolidine-4-carboxylic acid was formed. The reaction was inhibited by carbon monoxide showing the involvement of the cytochrome P-450-dependent mixed function oxidase system. The metabolism of carbon tetrachloride to phosgene may play a role in the production of hepatotoxicity by this compound.