Activation of the virion-associated RNA polymerase of vesicular stomatitis virus by melittin.
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Animals↗
Biomedical subjects
J Roy
Publications and source records attributed to J Roy.
Radio-labelling of 1-O-alkyl, 2-O-acetyl, sn-glycero-3-phosphorylcholine, 1-O-(9,10-di3H)-octadecyl PAF-acether.
3H-labelling of 1-3H-PAF-aceter was prepared by catalytic reduction of 1-O-(9,10)-octadecenyl-2-O-benzyl-sn-glyceryl-3-phosphorylcholine and then acetylated. 5 mumoles of this dehydro derivative dissolved in methanol was treated during 4 hours with tritium gas (80 Ci) and compressed until 1.1 bar by an automatic tritium transfer unit [1]. The catalyst was removed by filtration over Millex (Millipore) and labile tritium atoms eliminated by rotatory flash evaporation. The chemical purity was checked by 3H scanning and autoradiography. It was finally recovered through preparative thin-layer chromatography and then acetylated. The specific radioactivity was found to be close to 40-50 Ci/mmole.
Isotope Labeling↗
Biofunctional evaluation of a hydrogen bond stabilizing the conformation in the cyclic part of oxytocin.
[5-beta-Malamidic acid]oxytocin was synthesized to study the importance of the hydrogen bond between the C=O of Tyr2 and the peptide N-H of Asn5 for the stabilization of a biologically functional conformation of oxytocin. This analog lacks the peptide N-H at residue 5 required for the formation of a hydrogen bond with the C-O of Tyr2. [5-beta-Malamidic acid] oxytocin exhibited 45.1 +/- 2.5 U/mg and 65.6 +/- 5.9 U/mg of uterotonic activity, in vitro, in the absence and in the presence, respectively, of Mg2+, 147 +/- 14 U/mg of uterotonic activity in vivo, 203 +/- 13 U/mg of milk-ejecting activity, 0.37 +/- 0.03 U/mg of pressor activity and 0.32 +/- 0.29 U/mg of antidiuretic activity. It is concluded that devoid of the hydrogen bond under question, an oxytocin-like peptide can still assume the conformation needed to interact with the oxytocin receptors.
Drug Stability↗
Enkephalin dipeptidyl carboxypeptidase (enkephalinase) activity: selective radioassay, properties, and regional distribution in human brain.
The compound [3H]Tyr1,D-Ala2,Leu-OH5]enkephalin has been synthesised as a potentially selective substrate for enkephalin dipeptidyl carboxypeptidase (enkephalinase) activity in brain. Incubations in the presence of homogenates and particulate fractions from rodent and human brain result in the formation of [3H]Tyr-D-Ala-Gly, which can be conveniently isolated by polystyrene bead column chromatography. The enzyme activity responsible for the hydrolysis of the Gly3-Phe4 amide bond of this substrate displays close resemblance to that hydrolysing the natural enkephalins at the same level. In addition, enkephalinase activity characterised in postmortem human brain is closely similar to that in rodent brain, with regard to optimal pH and apparent affinities of various substrates and inhibitors, including the potent compound thiorphan. Enkephalinase activity is distributed in a highly heterogeneous fashion among regions of human brain, the highest levels being found in globus pallidus and pars reticulata of the substantia nigra. This distribution is poorly correlated with that of opiate receptor binding sites but displays some resemblance to that of reported Met5-enkephalin levels.
Aged↗
Enkephalin dipeptidyl carboxypeptidase ("enkephalinase"): assay, properties, and distribution in human brain and exploration of the active site.
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Binding Sites↗
Binding and metabolism studies with [3H](D-Ala2,Leu5)enkephalinamide and [3H](D-Ala2,Pro5)enkephalinamide: evidence for a selective interaction of (D-Ala2,Pro5)enkephalinamide with mu-receptors.
We have compared the binding characteristics of [3H](D-Ala2,Leu5)enkephalinamide and [3H](D-Ala2,Pro5)enkephalinamide on a washed homogenate from mouse brain. The maximum number of binding sites for [3H](D-Ala2,Leu5)enkephalinamide was twice that obtained with [3H](D-Ala2,Pro5)enkephalinamide. Hill slopes of the displacement curves for opiates and (D-Ala2,Pro5)enkephalinamide obtained against [3H](D-Ala2,Leu5)enkephalinamide were considerably lower than that of (D-Ala2,Met5)enkephalinamide and (D-Ala2,Leu5)enkephalinamide. In contrast, Hill slopes for morphine, ketocyclazocine and (D-Ala2,Leu5)enkephalinamide were similar for the displacement of [3H](D-Ala2,Pro5)enkephalinamide. These results are discussed and interpreted in terms of a selective interaction for (D-Ala2,Pro5)enkephalinamide with mu-receptors and a similar affinity of (D-Ala2,Leu5)enkephalinamide for mu- and delta-receptors. Since metabolism studies did not show any difference in the degradation rate of these two enkephalin analogs, the discrepancy between in vitro and in vivo activity cannot result from the catabolism processes. Another possible explanation is proposed.
Animals↗
Effects of traumatic manipulations on grafts, sutures, and host arteries during vascular surgery procedures. Experiments on dogs.
Scanning electron microscopy was used to diagnose incidents of trauma and the pattern of healing following surgical implantation of microporous blood vessel substitutes. Vascular reconstruction procedures using autogenous or synthetic prosthetic material inflict damage to the adjacent tissues as well as the prosthesis. This affects the thrombotic behavior and healing pattern of the area. The prognosis for long-term success are, as a consequence, directly influenced. Various vessel-occluding devices were evaluated in canine models with respect to their ability to damage blood vessels at the clamped site. The impact of suture needles on tissues and prosthetic materials was also considered. Of the various vessel-clamping devices which are commercially available, those which have elastomeric shields on the clamping components were found to be most satisfactory. The development of less traumatic surgical devices is discussed with the aim of further reducing surgical trauma.
Animals↗
Defibrinogenation as an alternative to heparinization in prolonged extracorporeal circulation.
Complications arising from difficulty controlled bleeding and thrombus formation during procedures which require extracorporeal circulation with heparin as anticoagulant motivate the search for better hemostasis and anticoagulant technology. An enzyme (Defibrase), having a specific interaction with fibrinopeptide A such as to cause fibrin depletion in a soluble form, has been proposed as an alternative to heparin. Defibrase, in contrast to heparin, does not affect blood platelet function. Heparin and Defibrase as anticoagulants were compared in 8 h perfusion studies using arterio-venous extracorporeal blood circuits in dogs; the circuitry included pumps, membrane oxygenators and filters serially. Thrombus formation, in the filters and the oxygenators as well as pathology of the perfused animal were investigated. The results suggest that defibrase-like enzymes have potential for enhanced control of hemorrhagic and thrombotic phenomena. Although impeded by a latency of several hours before the full anticoagulant properties are developed in vivo, it appears that enzymatically mediated defibrinogenation of blood may be a valuable alternative to heparinization in extracorporeal circulation procedures.
Animals↗
Studies on the importance of the asparagine residue in oxytocin. Synthesis and some pharmacological properties of [1-alpha-mercaptoacetic acid, 5-isoasparagine] oxytocin.
[1-Alpha-Mercaptoacetic acid, 5-isoasparagine] oxytocin was synthesized to study the effects of moving the side chain carboxamide group of the amino acid residue in position 5 of oxytocin from the beta to the alpha position. The analog has an isoasparagine residue in position 5 and the 20-membered ring size of oxytocin is maintained by substituting cysteine in position 1 of oxytocin by alpha-mercaptoacetic acid. The analog was found to possess 0.098 +/- 0.002 U/mg of uterotonic activity but no milk-ejecting, antidiuretic or rat pressor activity could be detected. The substance did not inhibit the uterotonic or milk-ejecting activity induced by oxytocin nor the antidiuretic or rat pressor responses to the USP posterior pituitary standard. These results, together with the data available in the literature, indicate that an analog of oxytocin lacking the asparagine residue in position 5 is neither an agonist nor an antagonist. The observations may mean that the asparagine residue is critically important for the interaction of oxytocin with its receptor.
Amino Acid Sequence↗
Biofunctional evaluation of a hydrogen bond stabilizing the beta-turn in the acyclic part of oxytocin.
In a continued effort to determine the importance of the hydrogen bonds for stabilization of the biologically active conformation of oxytocin, deamino-[9-glycolicamide] oxytocin was synthesized in order to study, in this respect, the hydrogen bond between the peptide N--H of Gly9 and the C=0 of Cys6. In this analog the amide linkage between residues at positions 8 and 9 is replaced by an ester. Thus the residue at position 9 cannot be involved in hydrogen bond formation with the C=O of Cys6. Deamino-[9-glycolicamide] oxytocin exhibited 134 +/- 13 U/mg and 355 +/- 48 U/mg of uterotonic activity in absence and in presence, respectively, of Mg2+, 108 +/- 8 U/mg of milk-ejecting activity, 0.35 +/- 0.03 U/mg of pressor activity and 2.5 +/- 0.1 U/mg of antidiuretic activity. It is concluded that the hydrogen bond under question is not critical for the conformation required for biofunctional interaction of oxytocin with its receptors in the uterus, mammary gland and other target organs.
Amino Acid Sequence↗
Prolonged responses by the rat uterus in vivo to deamino-oxytocin disproportionate to its estimated plasma half-life.
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Animals↗
[The treatment of fracture-dislocation of the humeral head by blind pinning (author's transl)].
The authors have treated 7 fracture-dislocations of the humeral head by blind pinning. The fractures were reduced by traction under general anaesthesia. 3 or 4 pins were then introduced into the olecranon fossa and driven up through the humeral head. After an average follow up of 29 months, the results were excellent in 9 cases, good in 5 and fair in 3. In 3 cases only, necrosis of the humeral head was noted.
Fracture Fixation, Internal↗
[Comparative study of anticoagulation methods available for extracorporeal circulation in the dog].
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Animals↗
Cardiovascular sutures as assessed by scanning electron microscopy.
Nine types of commercial sutures were investigated with respect to their suitability in cardiovascular surgery involving anastomosis of arterial prostheses. Polyester, nylon, silk and polypropylene in the form of multifilaments and monofilaments with or without coatings were included. The gross structure of the suture and the needle, the uniformity of the fibres and the coatings, the mechanical properties, before and after surgical use and the quality of the resulting anastomoses in canine models were evaluated. Although all surveyed products were potentially usable for the purpose intended, some appeared more suitable. Intraoperative damage to the sutures was found to be extensive and may play an important part in the long-term security of the anastomoses.
Cardiac Surgical Procedures↗
A SEM investigation of the trauma to prostheses and arteries during vascular reconstruction procedures.
Scanning electron microscopy was used to diagnose incidents of trauma and the pattern of healing following surgical implantation of microporous blood vessel substitutes. Vascular reconstruction procedures using autogenous or synthetic prosthetic materials inflict damage to the adjacent tissues as well as the prosthesis. This effects the thrombotic behavior and healing pattern of the area. The prognoses for long term success are, as a consequence, directly influenced. Various vessel occluding devices were evaluated on canine models with respect to their ability to damage blood vessels at the clamped site. The impact of suture needles on tissues and prosthetic materials was also considered. Of the various vessel clamping devices which are commercially available, those which have elastomeric shields on the clamping components were found to be the most satisfactory. The development of less traumatic surgical devices and possible modifications in the surgical protocols are discussed with the aim of further reducing surgical trauma.
Animals↗
Synthesis of [8-alpha-hydroxyisocaproic acid]oxytocin. Biological activities and comparison of its in vivo uterine actions to those of deamino-[8-alpha-hydroxyisocaproic acid]oxytocin, deamino-oxytocin and oxytocin.
The synthesis and biological activities of [8-alpha-hydroxyisocaproic acid]oxytocin are reported. The in vivo uterine responses to [8-alpha-hydroxyisocaproic acid] oxytocin are prolonged and those to deamino-[8-alpha hydroxyisocaproic acid] oxytocin (as well as those we have previously reported for deamino-oxytocin) are very prolonged as compared with those of oxytocin. Time courses of the in vivo uterine responses to deamino-[8-alpha-hydroxyisocaproic acid]oxytocin and deamino-oxytocin (as followed by plotting the intervals between the individual contractions of a response vs. time after peptide injection) differ from those to [8-alpha-hydroxyisocaproic acid]oxytocin and oxytocin.
Animals↗
Biofunctional evaluation of a hydrogen bond linking the ring and tail beta-turns of oxytocin.
Deamino-[8-N-methylleucine]oxytocin and deamino-[8-alpha-hydroxyisocaproic acid]oxytocin were synthesized to study the importance of hydrogen bonding between the carboxamide carbonyl of asparagine and the peptide N-H of leucine in stabilizing the biologically active conformation of oxytocin. The analogs were synthesized by coupling deaminotocinoic acid with Pro-Leu(Me)-Gly-NH2 and Pro-HyIc-Gly-NH2, respectively. (HyIc is alpha-hydroxyisocaproic acid). Deamino-[8-N-methylleucine]oxytocin was found to possess 48 +/- 7 units of uterotonic activity, 33 +/- 5 units of avian vasodepressor activity, and 3.15 +/- 1.5 units of antidiuretic activity per mg; deamino-[8-alpha-hydroxyisocaproic acid]oxytocin possessed 134 +/- 12 units of uterotonic activity, 31 +/- 3 units of avian vasodepressor activity, 9.6 +/- 3.0 units of antidiuretic activity, and 0.26 +/- 0.02 unit of pressor activity per mg. Neither of the analogs possesses the peptide N-H at residue 8 required for the formation of a hydrogen bond with the asparagine carboxamide; however, both can assume the conformation needed to evoke the characteristic biological activities of oxytocin although in lower potency. It is concluded that such a hydrogen bond does not constitute a conformational constraint that is essential for hormone action.
Animals↗
Conformational studies on [Pro3, Gly4]-oxytocin in dimethyl sulfoxide by 1H nuclear magnetic resonance spectroscopy: evidence for a type II beta turn in the cyclic moiety.
A model for oxytocin has been previously proposed in which residues 3 and 4 occupy the corner positions in a beta turn (Urry, D. W., & Walter, R (1971) Proc. Natl. Acad. Sci. U.S.A. 68, 956). The analogue [Pro(3),Gly(4)]-oxytocin (PGO) was used in proton magnetic resonance (1H NMR) studies designed to probe the contribution of these corner positions in forming a beta turn. Comparison of various 1H NMR parameters obtained at 220 MHz for backbone amide protons of PGO with those for the corresponding protons of oxytocin suggests that, to a first approximation, these two peptides may have similar backbone conformations in )CD3)2SO. Theoretically, the L-Pro lead toGly sequence in positions 3 and 4 of PGO would allow the formation of either a type I or type II beta turn. The two coupling constants between vicinal amide and C alpha protons for Gly(4) of PGO in (CD3)2SO are compatible with a type II beta turn in which 04, the dihedral angle about the bond between the backbone C alpha and N' atoms of Gly4, is appromximately +92 degrees, but not with a type I beta turn. A survey of peptides studied in other laboratories by X-ray and/or 1H NMR with reported type II beta turns with L-Pro lead toGly or Gly lead to Gly sequences in the corner positions revealed a close correlation between chemical shifts and vicinal coupling constants for the glycl residue in the second corner position. It is suggested that this criterion can form an additional basis for the characterization of beta turns. More studies are needed to determine the particular type of beta turn manifest in the cyclic moiety of oxytocin per se, although a reasonable working hypothesis is that oxytocin, similar to PGO, has a type II beta turn.
Dimethyl Sulfoxide↗