Cyclic hexa- and pentapeptide somatostatin analogues with reduced gastric inhibitory activity

The gastric inhibitory activity of cyclic hexa- and pentapeptide analogues of somatostatin was investigated in conscious cats with gastric fistulae. Gastric acid and pepsin secretions were stimulated by pentagastrin. Cyclo(Phe-Phe-D-Trp-Lys-Thr-Phe) showed no inhibition of acid secretion at molar doses up to 50-times the ID50 for somatostatin. This peptide inhibited pepsin secretion at the highest dose (50 micrograms kg-1 hr-1), and its potency is approximately 0.005 compared with somatostatin (1.0). Cyclo(Pro-Phe-D-Trp-Lys-Thr-Phe) inhibited acid (approximately 50%) and pepsin (approximately 85%) secretions, but the inhibition was not dose-related being similar with doses of 10 to 50 micrograms kg-1 hr-1. The cyclic pentapeptide, cyclo(7-aminoheptanoyl-Phe-D-Trp-Lys-Thr), was inactive in the dose range studied, with a potency less than 0.01. Cyclo[7-aminoheptanoyl-Phe-D-Trp-Lys-Thr(Bzl)] has been described as a somatostatin antagonist with respect to inhibition of growth hormone, insulin and glucagon release in rats [2]. Up to 60-fold molar excesses of this peptide failed to antagonise the inhibitory activity of somatostatin in the stomach. The results demonstrate that residues outside the central 6-11 region of somatostatin are very important for its gastric activity. The lack of gastric antagonistic activity of the pentapeptide antagonist indicates that these residues are likely to be involved in receptor recognition/binding.     

Circular dichroism and absorption study of the structure of methionine-enkephalin in solution

The circular dichroism and absorption spectra of methionine-enkephalin have been measured in aqueous solution, as functions of temperature and pH, and in 2,2,2-trifluoroethanol. Ranges covered were: 190–330 nm; 5–50°C; pH = 1–12. Absorption data provide no evidence for a strong intramolecular hydrogen bond involving the hydroxyl proton of the tyrosine residue. All data can be interpreted without assuming methionine-enkephalin preferentially occupies a single conformation when in dilute solution. The biologically important conformation is presumably one of many which are present to significant extent.     

Effects of HIS1 modifications on the ability of vasoactive intestinal peptide to stimulate adenylate cyclase from rat and human tissues

The importance of the N-terminal His residue of VIP for stimulating adenylate cyclase was appreciated by estimating the intrinsic activity and EC50 of four VIP analogues on membranes from rat lung, liver, brain, anterior pituitary, and pancreas, and on human heart membranes. In all tissue preparations tested except one, the order of efficacy (and often potency) was: VIP greater than (Ac-His1)VIP greater than (Phe1)VIP = (3-Me-His1)VIP greater than (D-His1)VIP. In rat heart membranes, the order of efficacy was somewhat different: VIP greater than (Ac-His1)VIP = (Phe1)VIP greater than (D-His1)VIP greater than (3-Me-His1)VIP. These data demonstrated the key role of His1 in VIP in activating adenylate cyclase. They suggest that a given VIP analogue might act as full agonist in tightly coupled adenylate cyclase systems (such as those of rat lung and liver membranes) whereas the same analogue could not promote full activity in poorly coupled systems (such as that present in rat brain synaptic membranes).     

SplitsTree: analyzing and visualizing evolutionary data

MOTIVATION: Real evolutionary data often contain a number of different and sometimes conflicting phylogenetic signals, and thus do not always clearly support a unique tree. To address this problem, Bandelt and Dress (Adv. Math., 92, 47-05, 1992) developed the method of split decomposition. For ideal data, this method gives rise to a tree, whereas less ideal data are represented by a tree-like network that may indicate evidence for different and conflicting phylogenies. RESULTS: SplitsTree is an interactive program, for analyzing and visualizing evolutionary data, that implements this approach. It also supports a number of distances transformations, the computation of parsimony splits, spectral analysis and bootstrapping.      

CNS effects of peripherally administered brain peptides.

Acceptance of enkephalins and endorphins into the family of brain peptides involves recognition that these endogenous opiates should share the general properties, including multiple and independent effects, previously described for neuropeptides. Several peptides first isolated by their pituitary-mediated endocrine effects, for example, are known to initiate CNS actions even in hypophysectomized animals. It was reasonable to expect, therefore, that the new opiate peptides would have effects not limited to the centrally induced analgesia by which they were originally identified, but, like the other brain peptides, would have additional CNS actions. Our concept that the multiple actions of peptides can be independent of each other is supported by evidence that even though peripheral administration of the brain opiates is essentially ineffective in producing analgesia, other actions of these peptides, such as changes in behavior, can be observed after administration by this route. Considerable evidence is accumulating in support of this concept of dissociation. The mechanisms by which the central effects of the peptides are exerted after systemic injection remain to be clarified, but analysis of their actions represents a new approach to understanding the performance of the brain. Studies already suggest a possible role of the brain peptides in the diagnosis and treatment of some mental and neurological disorders as well as in optimizing normal CNS functions.     

Evidence for direct production of somatostatin-14 from a larger precursor than somatostatin-28 in a phaeochromocytoma

Gel-filtration chromatography of an acid-extract of a phaeochromocytoma, under dissociating conditions, revealed 4 peaks of immunoreactive somatostatin (IRS) of approx. 8-10 kilodaltons (K), 6K, 3.5K and 1.6K as detected by an antiserum (R9) directed against the central region of tetradecapeptide somatostatin (S14). The 3.5K and 1.6K forms of IRS co-eluted with synthetic cyclic S28 and S14 respectively on reversed phase HPLC. Using another radioimmunoassay for the 1-14 sequence of S28 (N-peptide) a peak of immunoreactive N-peptide (IRN) with a molecular weight of approx. 4500 was observed. The antiserum (N3) used in the N-peptide assay was raised against N-Tyr N-peptide and cross-reacts less than 5% with synthetic S28. Two peaks were further characterised by partial tryptic digestion and gel-filtration chromatography. The 3.5K IRS peak was partially converted to a 1.6K IRS form together with an approximately equimolar amount of IRN with apparent molecular weight of 2500. This 2.5K IRN co-eluted both with N-Tyr N-peptide and with the IRN generated by tryptic digestion of synthetic cyclic S28. No IRN peak of this size was observed in the original extract. Tryptic digestion of the 6K IRS peak generated 3.5K and 1.6K IRS and 2.5K IRN. These results suggested that (1) this human phaeochromocytoma contains IRS very similar to the known structure of ovine and porcine S28 and S14. (2) The 6K IRS is composed of an unknown peptide sequence attached via trypsin-susceptible bond to the N-terminus of S28. (3) In this tumour S14 is being generated directly from 6K IRS and not via S28.     

Isolation, structure and synthesis of a heptapeptide with in vitro ACTH-releasing activity from porcine hypothalamus

Significant CRF activity was found in a fraction with Rf = 0.82-0.7 or VE/VT = 0.41-0.48 obtained by gel filtration of acid extracts of pig hypothalami on Sephadex G-25. The activity of this fraction decreased markedly during subsequent purification, particularly in the last two steps. From this fraction, a heptapeptide with significant ACTH releasing activity in vitro, was isolated in pure state, and its amino acid sequence was established as H-Phe-Ile-Tyr-His-Ser-Tyr-Lys-OH. This heptapeptide was synthesized by solid phase methods. The CRF activity of synthetic heptapeptide in vitro was low but could be potentiated by a cofactor fraction from rat hypothalamic extract.     

Presence and distribution of sensory structures on the mouthparts of self-medicating moths

Unique structures often accompany unusual feeding behaviors in erebid moths. We test whether self-medicating (e.g., pharmacophagous) adult tiger moths have specialized structures on their mouthparts. We examined mouthparts of pharmacophagous and non-pharmacophagous adults using scanning electron microscopy (SEM). Self-medicating adults had significantly higher numbers of chemosensory structures when compared with non-self-medicating adults. Putative olfactory sensilla are reported on the proboscis of Nyctemera coleta and pollen grains were found adhered to the proboscis of Nyctemera secundiana. Sensilla on the observed tiger moth proboscides may play a role in the recognition of pyrrolizidine alkaloid plants by pharmacophagous adults.