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Focuses on a study which assessed the effect of Smad4 and Apc genotypes in carcinogen-induced pancreatic lesions in the mouse. List of genes identified as mutated in pancreatic cancer; Information on adenomatous polyposis coli; Way in which the role of SMAD4 in colorectal tumor progression was established.

Defects of mismatch repair are thought to be responsible for carcinogenesis in hereditary non-polyposis colorectal cancer and about 15% of sporadic colon cancers. The phenotype is seen as microsatellite instability and is known to be caused either by mutations in mismatch repair genes or by aberrant methylation of these genes stabilizing their downregulation. Lack of repair of microsatellite sequence errors, created during replication, leads to a mutation-prone phenotype. Where mutations occur within mononucleotide tracts within exons they cause translation frameshifts, premature cessation of translation and abnormal protein expression. Such mutations have been observed in the TGFβRII, BAX, IGFIIR, MSH3 and MSH6 genes in colon and other cancers. We describe here frameshift mutations affecting the gene for the methyl-CpG binding thymine glycosylase, MBD4, in over 40% of microsatellite unstable sporadic colon cancers. The mutations all appear heterozygous but their location would ensure truncation of the protein between the methyl-CpG binding and glycosylase domains, thus potentially generating a dominant negative effect. It is thus possible that such mutations enhance mutation frequency at other sites in these tumours. A suggestion has been made that MBD4 (MED1) mutations may lead to an increased rate of microsatellite instability but this mechanism appears unlikely due to the nature of mutations we have found. [ABSTRACT FROM AUTHOR]

We wish to construct a mouse model for the human inherited disease cystic fibrosis. We describe here the successful targeting in embryonal stem cells of the murine homologue ( Cftr) of the cystic fibrosis transmembrane conductance regulator gene, as the first critical step towards this end. The targeting event precisely disrupts exon 10, the site of the major mutation in patients with cystic fibrosis. The targeted cells are pluripotent and competent to form chimaeras. [ABSTRACT FROM AUTHOR]

The derivation of a karyotypically normal embryonal stem (ES) cell line, E14, from inner cell masses (ICMs) isolated by immunosurgery from 129/Ola late mouse blastocysts is described. Disaggregated ICMs were cultured on mitotically-arrested fibroblast feeder layers in droplets of medium conditioned with Buffalo rat liver (BRL) cells under oil. BRL-conditioned medium inhibits the differentiation of established embryonal carcinoma (EC) and ES cell lines which can be maintained indefinitely in the complete absence of feeder cells (Smith and Hooper 1987). At clonal densities, however, a combination of BRL-conditioned medium and a feeder layer was most effective in preventing the differentiation of E14 cells. This effect was less pronounced at higher passage suggesting it may be particularly important to use a combination in the early stages of isolation. Once established, E14 has been maintained in BRL-conditioned medium alone. In non-conditioned medium on agarose, E14 cells formed embryoid bodies which when allowed to reattach differentiated into a wide variety of tissues. An HPRT-deficient sub line of E14, E14TG2a, has been demonstrated to form germline chimaeras with high efficiency after injection into blastocysts (Hooper et al. 1987). The modifications to the ES cell isolation procedure described here may improve the efficiency with which karyotypically normal lines can be derived. [ABSTRACT FROM AUTHOR]

Immunosurgical isolation of inner cell masses (ICMs) from sheep embryos was most efficient at the expanded, zona-intact blastocyst stage (day 7 to 8 post oestrus) before migration of endoderm cells beyond the boundary of the ICM across the blastocoelic surface of the trophectoderm. When cultured under conditions which allow the isolation of embryonal stem (ES) cell lines from mouse ICMs, sheep ICMs attached, spread and developed areas of both ES cell-like and endoderm-like cells. After prolonged culture only endoderm-like cells were evident. The implications for the isolation of ES cell lines from sheep embryos and possible species-specific requirements are discussed. [ABSTRACT FROM AUTHOR]

Airborne viable and total fungal spores were sampled inside and outside 80 houses in the Latrobe Valley, Victoria, Australia as part of a larger indoor environmental study. Each residence was visited six times over a period of 1 year for sample collection, and fungal spore samples were collected from at least three indoor sites and from an outdoor site. Viable spores were sampled using an Andersen sampler, while total spores were assessed using a Burkard spore trap. Identification of fungal colonies to genera level was performed in two seasons; winter and late spring. The most common fungal genera/groups were Cladosporium, Penicillium, and yeasts, both indoors and outdoors in winter and late spring. Outdoor levels were higher than those indoors throughout the year, and a significant seasonal variation in spore levels was seen both indoors and outdoors with overall maxima in summer. Contrary to this trend, the levels of Aspergillus, yeasts, Cephalosporium and Gliocladium were higher in winter. Most fungal genera were found in greater concentrations outdoors compared to indoors, but Penicillium was more common indoors. Outdoor spore levels were a significant influence on indoor levels, but seasonal differences suggest that other influences are important. [ABSTRACT FROM AUTHOR]

A flow injection method was developed for the determination of palladium, based on the reaction of palladium with DBOK-chlorophosphonazo with a detection limit of 0.07 μg/mL and a linear calibration range from 0.1 to 3.2 μg/mL of palladium. The reaction product has a maximum absorption at 630 nm. The method was applied to determine Pd2+ in catalysts and in anode mud samples. The relative error is less than 3% and the recovery of palladium is in the range 95% to 105% [ABSTRACT FROM AUTHOR]