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Transgenic rabbits carrying gene constructs encoding human nerve growth factor beta (hNGF‐β) cDNA were generated. Expression of hNGF‐β mRNA was restricted to the mammary gland of lactating rabbits. Western Blot analysis revealed a polypeptide of 13.2 kDa in the milk of transgenic animals. hNGF‐β was purified from the milk by a two‐step chromatographic procedure. Electrospray mass spectroscopy analysis of purified hNGF‐β depicted a molecular weight of 13 261 Da per subunit. The biological activity of the hNGF‐β was tested using PC12W2 cells and cultures of dorsal root ganglion neurons from chicken embryos. Crude defatted milk from transgenic animals and purified hNGF‐β demonstrated full biological activity when compared to commercial recombinant hNGF‐β.

A complementary DNA (cDNA) encoding a serotonin receptor with 51% sequence identity to the 5HT‐1C subtype was isolated from a rat brain cDNA library by homology screening. Transient expression of the cloned cDNA in mammalian cells was used to establish the pharmacological profile of the encoded receptor polypeptide. Membranes from transfected cells showed high‐affinity binding of the serotonin antagonists spiperone, ketanserin and mianserin, low affinity for haloperidol (a dopamine D2 receptor antagonist), 8‐OH‐DPAT as well as MDL‐72222 and no detectable binding of [3H]serotonin. This profile is consonant with the 5HT‐2 subtype of serotonin receptors. In agreement with this assignment, serotonin increased the intracellular Ca2+ concentration and activated phosphoinositide hydrolysis in transfected mammalian cells. The agonist also elicited a current flow, blocked by spiperone, in Xenopus oocytes injected with in vitro synthesized RNA containing the cloned nucleotide sequences.

The rate of convergence of a multidimensional random search is found to improve when the search region is restricted to a directed adaptive hypercone. The convergence properties of the hyperconical search algorithm are investigated and the scheme is applied to the problem of identifying twenty-two constants of a given chemical reaction from measured data.

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