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The objective of this study was to examine the degradation of short peptides corre­sponding to modified fragments of human growth hormone-releasing hormone by trypsin. Six analogues of pentapeptide 9-13 of human growth hormone-releasing hor­mone containing homoarginine, ornithine, glutamic acid, glycine, leucine or phenyl­alanine residue in position 11, two analogues of hexapeptide 8-13 of human growth hormone-releasing hormone and two analogues of heptapeptide 7-13 of human growth hormone-releasing hormone containing homoarginine or glycine residue in position 11 were obtained. The peptides were subjected to digestion by trypsin and the course of reaction was monitored using HPLC. It was found that the rate of hy­drolysis of the Lys12-Val13 peptide bond depends on the amino-acid residue preceding Lys12 . The extension of the peptide chain towards the N-terminus by introduction of consecutive amino-acid residues corresponding to the human growth hormone-re­leasing hormone sequence accelerates the hydrolysis process. These results may be of assistance in designing new analogues of human growth hormone-releasing hor­mone, more resistant to the activity of proteolytic enzymes.
Conformational space of a novel cyclic enkephalin analogue, cyclo(Nε, Nε'-carbonyl-D-Lys2,Lys5) enkephalinamide, was exhaustively examined. A large number of conformations was selected and clustered into families on the basis of their structure and energy. For representative conformations ROESY spectra were generated and their linear combination was fitted to the spectra measured in water and Me2-SO-d6. This procedure yielded an ensemble of most populated conformations of the peptide in the two solvents.
Delta opioid receptors are involved in modulation of nociception, thermoregulation, locomotor activity and rewarding properties of drugs of abuse. The new analog of deltorphin – DK-4 is a peptide with high affi nity to delta opioid receptors in mouse vas deference bioassay. The aim of our study was to indicate whether delta opioid receptors are involved in the rewarding effects of cocaine. Therefore, we tested infl uence of (1) opioid receptor antagonists (μ, κ, δ) on the expression of cocaine-induced conditioned place preference (CPP); and (2) infl uence of DK-4 on the cocaine reinstatement of CPP (Sleipness et al. 2007). Rats were conditioned daily with cocaine (5 mg/kg, i.p.), tested on the 6th day (the expression of CPP), and then retested for 9 days to monitor the extinction of the CPP. On the day 10 animals were treated with a single injection of cocaine (5 mg/kg) or DK-4 (5, 10 nmol, i.c.v.) to reinstate the CPP. Our results indicated that naltrindole – a delta opioid receptor antagonist decreased cocaine rewarding effect (the expression of CPP). DK-4 increased the reinstatement of cocaine-induced CPP. These studies suggest that delta opioid receptors are involved in the cocaine-induced rewarding effects.
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The aim of our study was to check the responsiveness the chemoreceptor reflex in 28 young mildly hypertensive men (HTS), aged 18-32 years and 25 normotensive male subjects (NTS) aged 19-32 years, before and after 3-months dynamic exercise training. We tested the hypothesis that dynamic training reduces arterial chemoreceptor drive in mild hypertension. Circulatory response to 3-min hyperoxic inactivation of arterial chemoreceptors induced by 70% oxygen breathing was measured before and after training. Arterial blood pressure (BP) was recorded continuously by Finapres method, stroke volume and arm blood flow were registered by impedance reography, heart rate by ECG. Both groups were submitted to moderate 3-months dynamic exercise training. Before training the hyperoxic breathing caused in HTS a significant decrease in systolic BP by 6±1mmHg p<0.01, in diastolic BP by 2±0.6mmHg p<0.01, and in total peripheral vascular resistance (TPR) by 0.24±0.04 TPRU (p<0.01). After training hyperoxia augmented systolic BP by 2.64±1.9mmHg (NS), diastolic BP by 2±1mmHg p<0.05, and TPR by 0.043±0.05 TPRU (ANOVA). In NTS before training brief hyperoxia produced insignificant change in BP and TPR. In NTS after training hyperoxia increased systolic BP by 4.2 mm Hg±1.23 p<0.01 and diastolic BP by 3.1±0.6mmHg p<0.01 respectively and TPR by 0.053±0.02 TPRU. Our results confirm earlier finding on the enhanced arterial chemoreceptor reflex drive in mild human hypertension. We conclude that normalizing arterial blood pressure in subjects with mild hypertension which occurred after 3-months dynamical exercise training is due to attenuation of the sympathoexcitatory chemoreceptor reflex drive by exercise training. The mechanism of this effect requires further study.
The objective of our study was to compare the cardiovascular effects of moderate exercise training in heathy young (NTS, n=18, 22.9±0.44 years) and in hypertensive human subjects (HTS, n=30, 23±1.1). The VO2max did not significantly differ between groups. HTS of systolic blood pressure (SBP) 148±3.6 mmHg and diastolic blood pressure(DBP) 88±2.2mmHg, and NTS of SBP: 128.8 ± 4 mmHg and DBP: 72 ± 2.9 mmHg were submitted to moderate dynamic exercise training, at about 50% VO2max 3 times per week for one hour, over 3 months. VO2max was measured by Astrand's test. Arterial blood pressure was measured with Finapres technique, the stroke volume, cardiac output and arm blood flow were assessed by impedance reography. Variability of SBP and pulse interval values (PI) were estimated by computing the variance and power spectra according to FFT algorithm. After training period significant improvements in VO2max were observed in NTS- by 1.92 ±0.76 and in HTS by 3±0.68 ml/kg/min). In HTS significantly decreased: SBP by 19 ±2.9 mmHg, in DBP by 10.7±2 mmHg total peripheral resistance (TPR) by 0.28 ±0.05 TPR units. The pretraining value of low frequency component power spectra SBP (LFSBP) was significantly greater in HTS, compared to NTS. PI variance was lower in HTS, compared to NTS. After physical training, in HTS PI variance increased suggesting a decrease in frequency modulated sympathetic activity and increase in vagal modulation of heart rate in mild hypertension. A major finding of the study is the significant decrease of resting low frequency component SBP power spectrum after training in HTS. The value of LFSBP in trained hypertensive subjects normalized to the resting level of LFSBP in NTS. Our findings suggest that antihypertensive hemodynamic effects of moderate dynamic physical training are associated with readjustment of the autonomic cardiovascular control system.
 Six hybrid N-ureidoethylamides of octapeptides in which an N-terminal cyclic structure related to enkephalin was elongated by a C-terminal fragment of deltorphin were synthesized on MBHA resin. The synthetic procedure involved deprotection of Boc groups with HCl/dioxane and cleavage of the peptide resin with 45 % TFA in DCM. d-Lys and d-Orn were incorporated in position 2, and Lys, Orn, Dab, or Dap in position 5. The side chains of the dibasic amino function were protected with the Fmoc group. This protection was removed by treatment with 55 % piperidine in DMF, and cyclization was achieved by treatment with bis-(4-nitrophenyl)carbonate. Using various combinations of dibasic amino acids, peptides containing a 17-, 18-, 19- or 20-membered ring structure were obtained. The peptides were tested in the guinea-pig ileum (GPI) and mouse vas deferens (MVD) assays. Diverse opioid activities were observed, depending on the size of the ring. Extension of the enkephalin sequence at the C-terminus by a deltorphin fragment resulted in a change of receptor selectivity in favor of the δ receptor. The conformational propensities of selected peptides were determined using the EDMC method in conjunction with data derived from NMR experiments carried out in water. This approach allowed proper examination of the dynamical behavior of these small peptides. The results were compared with those obtained earlier with corresponding N-(ureidoethyl)pentapeptide amides.
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