No abstract

Oncogenic Ras proteins transform animal cells to a malignant phenotype only when modified by farnesyl residues attached to cysteines near their carboxyl termini. The farnesyltransferase that catalyzes this reaction recognizes tetrapeptides of the sequence CAAX, where C is cysteine, A is an aliphatic amino acid, and X is a carboxyl-terminal methionine or serine. Replacement of the two aliphatic residues with a benzodiazepine-based mimic of a peptide turn generated potent inhibitors of farnesyltransferase [50 percent inhibitory concentration (IC50) < 1 nM]. Unlike tetrapeptides, the benzodiazepine peptidomimetics enter cells and block attachment of farnesyl to Ras, nuclear lamins, and several other proteins. At micromolar concentrations, these inhibitors restored a normal growth pattern to Ras-transformed cells. The benzodiazepine peptidomimetics may be useful in the design of treatments for tumors in which oncogenic Ras proteins contribute to abnormal growth, such as that of the colon, lung, and pancreas.

Derivatives of human growth hormone (hGH) of increasing size were produced by reaction with the Nhydroxysuccinimide ester of polyethylene glycol-5000 (PEG 5000 ), a 5-kDa reagent that selectively conjugates to primary amines. By adjusting the reaction conditions and purification procedure, it was possible to isolate hGH derivatives containing up to seven PEG moieties that altered the Stokes radius and thereby the effective molecular masses of the unmodified hormone from 22 to 300 kDa. Fortunately, the most reactive amines were ones that did not lie in either of the two sites important for receptor binding. Nonetheless, increasing the level of PEG modification linearly reduced the affinity of hGH for its receptor and increased the EC 50 in a cellbased assay up to 1500-fold. Most of the reduction in affinity was the result of slowing the association rate for the receptor. The clearance rate of hGH in rats was inversely proportional to effective molecular weight and closely fit a filtration model. We have tested the potency of these analogs by injecting them daily or every 6 days into hypophysectomized rats and determining the effects on body and organ growth. The efficacy of these analogs was optimal for hGH conjugated with 5 eq of PEG 5000 , and the potency was increased by about 10-fold compared with unmodified hGH. Such PEG-hGH derivatives show promise as long-acting alternatives to daily injections of hGH. More generally these studies show that improving hormone clearance properties, even at the expense of reducing receptor binding affinity, can lead to dramatic increases in hormone efficacy.The ability of a hormone to elicit a biological effect in vivo depends on many factors including the affinity for its receptor and the rate at which it is cleared from the circulation. Some hormones, like atrial natriuretic peptide, have a very high affinity for their receptor (10 pM) and are cleared very rapidly (t1 ⁄2 ϳ0.5 min) by receptor and protease-mediated events (1). Other hormones, like human growth hormone (hGH), 1 have lower affinity for their receptor (300 pM) but are cleared more slowly (t1 ⁄2 ϳ30 min in rats), primarily via the kidney (2, 3).Understanding the relationships between hormone affinity, clearance, and efficacy is important in optimizing hormone therapy. To study this systematically one would like to vary these parameters and evaluate their relative importance in regulating biopotency. hGH is a good model system in this regard as much is known about its structure and function (for review see Ref. 4). Simple receptor binding (5, 6), cell-based assays (7,8), and growth parameters in rodents (9) can be used to determine biopotency in vitro and in vivo. The properties of proteins such as hGH that are cleared by kidney filtration can be modulated by attachment of polyethylene glycol (PEG) polymers, which increases the hydrodynamic volume of the hormone and thereby slows its clearance (10, for recent review see Ref. 11).Here, we describe a set of hGH derivatives conjugated with increasing numbers of PEG ...

No abstract

Following an exhibition of 32 current vintage red wines of two varieties from the Southern Vales district of South Australia, a correlation was observed between colour densities and the order of ranking previously assigned by a panel of experienced judges.There was no relation between wine colour density and anthocyanin content. Amongst many wines having comparable levels of anthocyanins, the variation in colour density was as much as 3.6-fold, and the degree of ionisation of the anthocyanin component of wine colour was found to range from 6 to 25%. These latter values were also correlated with the quality ratings.The data support the opinion that measurement of colour density and of the degree of ionisation of anthocyanins can provide an objective guide to organoleptic properties in young wines of the same variety and region. Confirmation for this view was obtained from other data concerning the subsequent vintage of Cabernet Sauvignon in the same district.Factors likely to be responsible for the varying states of anthocyanin equilibria in these wines are discussed in terms of wine composition and oenological practice.

No abstract

Whereas high levels of flavonoid extractives are responsible for the characteristic A,,, at about 280nm in the ultra-violet spectra of red wines, there is wide variability in the spectral features of natural white wines, in which the phenolic content is often minimal. For the latter, A,,, may occur anywhere within the range 265-285 nm, with relatively strong absorbance at 320 nm arising from the presence of hydroxycinnamate esters. Massive fining of many white wines and juices with polyvinylpyrrolidone (10% w/v) has shown residual non-phenolic U.V. absorbing constituents to be sufficiently uniform to enable direct spectral measurement of total hydroxycinnamates in white juices, white wines and roses as (E320-1 .4) absorbance units. For white grape juices and white wines, total hydroxycinnamates are directly quantitated as mg litre-' 'caffeic acid equivalents' and total flavonoid extractives may also be estimated. Total phenolic components of red wines are accounted as (Ez80-4) absorbance units. The spectral interpretations enable comparative estimation of flavonoid extractives in all natural wine types.

No abstract