Cycloheximide-sensitive synthesis of substance P by isolated dorsal root ganglia

Substance P (Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2) may be used as a neurotransmitter by certain primary afferent neurones, particularly those carrying pain impulses. Substance P-like immunoreactivity has been localised to the cell bodies of one population of dorsal root ganglion neurones by immunocytochemistry. It is contained in vesicles in the central terminals of these neurones, and has also been demonstrated in the peripheral terminals. As axons and terminals have very little capacity for peptide biosynthesis, it is possible that substance P is synthesised and packaged in the perikaryon and transported to the terminals by an axoplasmic transport process. Consistent with this is the finding that substance P accumulates proximal to a ligature placed on the dorsal root. There has, however, been no direct demonstration of the biosynthesis of substance P in the nervous system. We report here that rat dorsal root ganglia incorporate 35S-methionine into substance P, characterised as authentic by immunoprecipitation followed by HPLC. There is a delay of 1-2 h between addition of label and its incorporation into substance P. Synthesis is blocked by cycloheximde suggesting that, in dorsal root ganglia, substance P is synthesised by a conventional ribosomal process. Synthesis of substance P is reduced by some 90% in ganglia from rats treated neonatally with capsaicin, a drug which is thought to destroy a population of primary afferent neurones.     

Changes in high density lipoprotein subfractions during alimentary lipaemia

Summary Significant changes in high density lipoprotein subfractions accompanied alimentary lipaemia in normal subjects. Results emphasized the importance of using fasting subjects in HDL studies but did not support an in vivo transformation of HDL₃ to HDL₂.     

Novel thioether bond revealed by a 1.7 A crystal structure of galactose oxidase

Galactose oxidase is an extracellular enzyme secreted by the fungus Dactylium dendroides. It is monomeric, with a relative molecular mass of 68,000, catalyses the stereospecific oxidation of a broad range of primary alcohol substrates and possesses a unique mononuclear copper site essential for catalysing a two-electron transfer reaction during the oxidation of primary alcohols to corresponding aldehydes. Recent evidence arguing against a Cu(III)-Cu(I) couple implies the existence of a second redox-active site proposed to involve pyrroloquinoline quinone or a tyrosine radical. We now report the crystal structure of galactose oxidase at 1.7 A resolution. This reveals a unique structural feature at the copper site with a novel thioether bond linking Cys 228 and Tyr 272 in a stacking interaction with Trp 290. We propose that these molecular components stabilize the protein free-radical species essential for catalysis and thus provide a 'built-in' secondary cofactor. This feature may represent a new mechanism for mediating electron transfer in metalloenzymes in the absence of exogenous cofactors.     

Evidence of two temperate episodes in late Pleistocene deposits at Marsworth, UK

In the British Quaternary, two post-Cromerian interglacials, the Hoxnian and the Ipswichian, are recognized. Evidence of additional interglacials in this interval is widely accepted in the oceanic record of Quaternary events, and the possibility that at least one additional interglacial of this age is represented in Britain has been discussed. However, in the absence of datable interglacial deposits which are seen to overlie one another, the issue has remained controversial. We describe here deposits at Marsworth, UK (Fig. 1) where there is evidence of two temperate episodes, and of intervening periglacial conditions. Stratigraphical superposition is established beyond any reasonable doubt. The later deposit relates to the temperate woodland stage of the Ipswichian Interglacial. Dating of the earlier temperate material by the 230Th/234U disequilibrium method indicates an interglacial episode not previously established in the British Quaternary.     

A chronological framework for the British Quaternary based on Bithynia opercula

Marine and ice-core records show that the Earth has experienced a succession of glacials and interglacials during the Quaternary (last ～2.6 million years), although it is often difficult to correlate fragmentary terrestrial records with specific cycles. Aminostratigraphy is a method potentially able to link terrestrial sequences to the marine isotope stages (MIS) of the deep-sea record. We have used new methods of extraction and analysis of amino acids, preserved within the calcitic opercula of the freshwater gastropod Bithynia, to provide the most comprehensive data set for the British Pleistocene based on a single dating technique. A total of 470 opercula from 74 sites spanning the entire Quaternary are ranked in order of relative age based on the extent of protein degradation, using aspartic acid/asparagine (Asx), glutamic acid/glutamine (Glx), serine (Ser), alanine (Ala) and valine (Val). This new aminostratigraphy is consistent with the stratigraphical relationships of stratotypes, sites with independent geochronology, biostratigraphy and terrace stratigraphy. The method corroborates the existence of four interglacial stages between the Anglian (MIS 12) and the Holocene in the terrestrial succession. It establishes human occupation of Britain in most interglacial stages after MIS 15, but supports the notion of human absence during the Last Interglacial (MIS 5e). Suspicions that the treeless 'optimum of the Upton Warren interstadial' at Isleworth pre-dates MIS 3 are confirmed. This new aminostratigraphy provides a robust framework against which climatic, biostratigraphical and archaeological models can be tested.