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Domestication is a long-term process during which wild fish are acclimated to farming conditions and hopefully are reproduced over several generations, possibly using selective breeding. Preservation of the genetic diversity of the original population, together with that of the ongoing selection steps, is important for ecological and economical purposes. Cryobanking of reproductive cells is one answer to meet this need. In fish, however, only sperm can be cryopreserved as neither oocytes nor embryos are capable of handling the freeze-thawing stress. In this review, we explore to what extent diploid cells obtained from fin pieces can be used for the preservation of both parental genomes. The main parameters, which should be under control to ensure proper production of fin cells in culture and to enable cryopreservation of the material are described. After cryobanking of such non-reproductive cells, fish can be reconstructed using the nuclear transfer technology whose potentials and difficulties are discussed. The gametes produced by the fish reconstructed after somatic cells nuclear transfer are different to some extent from the gametes obtained after the direct transplantation of primordial germ cell or spermatogonial germ cells into host embryos or larvae. However, in some cases, only somatic cells can be obtained in quantities which would be compatible with strain restoration purposes. From the knowledge available today, it is reasonable to expect that nuclear transfer becomes available for fish reconstruction, even if restricted to high-tech biotechnology facilities. Therefore, cryobanking of fin somatic cells can be farsightedly considered for high-throughput diploid genome conservation. [ABSTRACT FROM AUTHOR]

Background: Nuclear transfer has the potential to become one strategy for fish genetic resources management, by allowing fish reconstruction from cryopreserved somatic cells. Survival rates after nuclear transfer are still low however. The part played by unsuitable handling conditions is often questioned, but the different steps in the procedure are difficult to address separately. In this work led on goldfish (Carassius auratus), the step of somatic cells injection was explored. Non-enucleated metaphase II oocytes were used as a template to explore the toxicity of the injection medium, to estimate the best location where the cell should be injected, and to assess the delay necessary between cell injection and oocyte activation. Results: Trout coelomic fluid was the most suitable medium to maintain freshly spawned oocytes at the metaphase II stage during oocyte manipulation. Oocytes were then injected with several media to test their toxicity on embryo development after fertilization. Trout coelomic fluid was the least toxic medium after injection, and the smallest injected volume (10 pL) allowed the same hatching rates as the non injected controls (84.8% ± 23). In somatic cell transfer experiments using non enucleated metaphase II oocytes as recipient, cell plasma membrane was ruptured within one minute after injection. Cell injection at the top of the animal pole in the oocyte allowed higher development rates than cell injection deeper within the oocyte (respectively 59% and 23% at mid-blastula stage). Embryo development rates were also higher when oocyte activation was delayed for 30 min after cell injection than when activation was induced without delay (respectively 72% and 48% at mid-blastula stage). Conclusions: The best ability of goldfish oocytes to sustain embryo development was obtained when the carrier medium was trout coelomic fluid, when the cell was injected close to the animal pole, and when oocyte activation was induced 30 min after somatic cell injection. Although the experiments were not designed to produce characterized clones, application of these parameters to somatic cell nuclear transfer experiments in enucleated metaphase II oocytes is expected to improve the quality of the reconstructed embryos. [ABSTRACT FROM AUTHOR]

Extracellular matrix metalloproteinase inducer (EMMPRIN) was examined on bronchoalveolar lavage fluids (BALFs) and lung tissue from patients with fibrosis (usual interstitial pneumonia–idiopathic pulmonary fibrosis [UIP-IPF], n = 15; diffuse parenchymal lung diseases without IPF characteristics on computerized tomography scan, n = 8) and without fibrosis (n = 6). In UIP-IPF, EMMPRIN staining was increased in areas of fibrosis, mainly in macrophages and in epithelial cells. EMMPRIN was also found in the extracellular medium with significant levels in patients with lung fibrosis compared to subjects without fibrosis. Moreover, macrophages from patients with lung fibrosis spontaneously produce EMMPRIN. These findings show that EMMPRIN is increased in lung fibrosis. [ABSTRACT FROM AUTHOR]

A solid-phase route for the preparation of 4a,5,8,8a-tetrahydrophthalazinon- 1-ones employing the Diels-Alder reaction has been developed. Some of the new compounds have been tested for inhibition of LPS-stimulated TNF-α production in human whole blood from patients with chronic obstructive pulmonary disease (COPD). This evaluation revealed two compounds 17 and 18 of interest, incorporating an arylpiperazine moiety, which were found to inhibit LPS- induced TNF-α release like the well known anti-inflammatory PDE4 inhibitors, rolipram and roflumilast. [ABSTRACT FROM AUTHOR]