Expanding the scope of methylation-sensitive restriction enzyme (MSRE) PCR for forensic identification of body fluids through the novel use of methylation-dependent restriction enzymes (MDRE) and the combination of autosomal and Y-chromosomal markers.

Methylation-sensitive/-dependent restriction enzyme (MSRE/MDRE) PCR can be performed to detect hypomethylated or hypermethylated CpG sites. With the combined use of different tissue-specific CpG markers, MSRE/MDRE-PCR leads to tissue-specific methylation patterns (TSMPs), enabling the correlation of DNA samples to their source tissue. MSRE/MDRE assays can use the same platform as forensic STR typing and offer many advantages in the field of forensic body fluid detection.

Mechanism of kinin release during experimental acute pancreatitis in rats: evidence for pro- as well as anti-inflammatory roles of oedema formation.

1 Kinin B(2) receptor antagonists or tissue kallikrein (t-KK) inhibitors prevent oedema formation and associated sequelae in caerulein-induced pancreatitis in the rat. We have now further investigated the mechanism of kinin generation in the pancreas. 2 Kinins were elevated in the pancreatic tissue already before oedema formation became manifest.

Severe hypoxia inhibits prostaglandin I(2) biosynthesis and vasodilatory responses induced by ionophore A23187 in the isolated rabbit ear.

The present study was designed to test the hypothesis that lack of oxygen in severely hypoxic tissue may inhibit arachidonic acid oxygenation and thereby result in an inhibition of eicosanoid synthesis. Hypoxia was induced in the isolated rabbit ear, and arachidonate metabolism and peripheral resistance of the preparation were monitored simultaneously. Severe hypoxia completely inhibited the biosynthesis of prostaglandin I(2) induced by ionophore A23187 and converted the vasodilatory response observed under normoxia into vasoconstriction.

Different modes of action of morphine and epibatidine in Straub tail reactions in mice.

The Straub tail reaction in mice is an S-shaped dorsiflexion of the mouse tail, long known as a sensitive and specific bioassay for morphine (CAS 57-27-2). It is based on a contraction of the sacro-coccygeal dorsalis muscles. In the case of morphine the reaction is thought to be induced by a long-lasting stimulation of the muscle's motor innervation at the level of the lumbosacral spinal cord.

Epibatidine: high potency and broad spectrum activity on neuronal and neuromuscular nicotinic acetylcholine receptors.

The activity of epibatidine at neuronal and neuromuscular nicotinic acetylcholine receptors was investigated in several in situ and in vitro systems and compared with the activity of nicotine and suxamethonium. Activation of ganglionic nicotinic receptors by epibatidine was shown in the guinea-pig ileum (contraction mediated by the cholinergic neurons of the ileum) and in pithed and atropinized rats (rise in blood pressure). Epibatidine also activated nicotinic receptors at the peripheral terminals of afferent C-fibres (rabbit ear) and in the brain (antidiuresis in rats).

In vivo effects of the NMDA receptor agonist tetrazolyl-glycine related to the function of small-diameter primary afferents.

The NMDA receptor agonist tetrazolyl-glycine (TG; 100 microg kg(-1), i.v.) caused a depressor reflex in anaesthetized rats.

Vespula vulgaris venom: role of kinins and release of 5-hydroxytryptamine from skin mast cells.

Wasp venoms contain several active components, among them kinin-related peptides. Like bradykinin and [Thr6]bradykinin, Vespula vulgaris venom caused paw oedema following subplantar injection in anaesthetized rats. The oedema was partly inhibited by the bradykinin B2 receptor antagonist icatibant (Hoe 140); the remaining part was abolished by additional pretreatment with 5-hydroxytryptamine (5-HT) receptor antagonists or mast cell depletion.

Afferent C-fibres release substance P and glutamate.

Capsaicin is currently used as a specific pharmacological tool for investigation of functions of primary afferent C-fibres. Their peripheral terminals play an important role in "neurogenic inflammation," mediated by released substance P and calcitonin gene related peptide, whereas in the mediation of central functions, activation of the N-methyl-D-aspartate (NMDA) receptors has recently been demonstrated. A method for continuous monitoring of glutamate concentration was used to study mechanisms of capsaicin-induced glutamate release from rat spinal cord slices.

Pathophysiological and possible physiological roles of kinins in the pancreas.

The i.v. infusion of a low dose of the cholecystokinin agonist caerulein elicited a sustained secretion of amylase into the biliopancreatic duct of rats.

Evidence for the presence of NK1 and NK3 receptors on cholinergic neurones in the guinea-pig ileum.

In a guinea-pig ileum longitudinal muscle preparation, substance P (SP) (> or = 6 nM) caused an initial contraction followed by a sustained plateau contraction of about 20-50% of the initial response. This plateau contraction is caused by the SP-induced activation of cholinergic motoneurones which contract the smooth muscles by the released acetylcholine (ACh). We investigated the contribution of neurokinin NK1 and NK3 receptors during this 'plateau phase' of contraction.

Evidence for the participation of glutamate in reflexes involving afferent, substance P-containing nerve fibres in the rat.

1. Responses mediated, either peripherally or centrally, by substance P-containing primary afferent C-fibres were investigated in the rat following impairment of axonal transport by colchicine (120 micrograms kg-1, i.p.

Effects of the bradykinin antagonist, icatibant (Hoe 140), on pancreas and liver functions during and after caerulein-induced pancreatitis in rats.

It has been found earlier that the bradykinin antagonist, icatibant (Hoe 140), prevents the pancreatic oedema and the ensuing hypotension and haemoconcentration, and facilitates the removal of activated enzymes form the tissue during caerulein-induced acute pancreatitis. For a potential therapeutic use of the compound in clinical situations it is essential to investigate whether the associated increase in enzyme activities in the blood serum has any adverse effects on the pancreas itself or on other organs. Normal amylase secretion into the biliopancreatic duct stimulated by a low dose of caerulein (0.

Anti-inflammatory and analgesic activity of the bradykinin antagonist, icatibant (Hoe 140), against an extract from Porphyromonas gingivalis.

1. Porphyromonas gingivalis is one of the bacteria likely to be related to pain in periodontitis. Several enzymes isolated from P.

Mediation by bradykinin of rat paw oedema induced by collagenase from Clostridium histolyticum.

1. Collagenases are thought to play a major role in the pathology of gas gangrene caused by Clostridium histolyticum, because they can destroy the connective tissue barriers. We investigated possible mediators involved in the oedema formation and plasma protein extravasation which follow the injection of a collagenase (EC 3.

Pathological events in experimental acute pancreatitis prevented by the bradykinin antagonist, Hoe 140.

1. In a previous investigation, Hoe 140, a specific and potent bradykinin B2 receptor antagonist, prevented the pancreatic oedema and the hypotension observed during acute experimental pancreatitis; however, it augmented the associated rises in the serum activities of pancreatic enzymes. Therefore, we have now investigated the consequences of the pancreatic oedema for the fate of activated enzymes released into the tissue during the course of acute pancreatitis.

Effects of the bradykinin antagonist, HOE 140, in experimental acute pancreatitis.

1. The novel bradykinin antagonist, HOE 140, completely blocked the fall in rabbit blood pressure caused, not only by i.v.

CP-96,345, a non-peptide antagonist of substance P. III. Cardiovascular effects in mammals unrelated to actions on substance P receptors.

The cardiovascular effects of CP-96,345, a non-peptide antagonist of substance P, were analyzed in vivo and in vitro. In the anaesthetized rat, the i.v.

CP-96,345, a non-peptide antagonist of substance P: II. Actions on substance P-induced hypotension and bronchoconstriction, and on depressor reflexes in mammals.

In order to extend the in vivo pharmacological profile of CP-96,345 ((2S,3S)-cis-2-(diphenylmethyl)-N-((2-methoxyphenyl)- methyl)-1-azabicyclo[2.2.2]octan-3-amine dihydrochloride), a non-peptide antagonist of substance P, the compound was tested against substance P-induced effects on blood pressure in rabbits and on respiration pressure in guinea-pigs.

CP-96,345, a non-peptide antagonist of substance P: I. Effects on the actions mediated by substance P and related tachykinins on the guinea-pig ileum and rabbit jejunum.

The effects of a non-peptide antagonist of substance P, CP-96,345, were investigated, in vitro, on the guinea-pig ileum and the rabbit jejunum. Contractions of the guinea-pig ileum, induced by substance P and neurokinin A, were specifically inhibited by the racemate (+/-)CP-96,345 (pIC50 7.8 and 7.

The non-peptide tachykinin antagonist, CP-96,345, is a potent inhibitor of neurogenic inflammation.

1. Release of the tachykinin, substance P, from the peripheral terminals of polymodal afferent C-fibres is thought to be largely responsible for the vasodilatation and plasma protein extravasation described as neurogenic inflammation. The effects of CP-96,345, a non-peptide antagonist at the substance P (NK1) receptor, on these vascular reactions were investigated in the rat.

Analysis of the antagonistic actions of HOE 140 and other novel bradykinin analogues on the guinea-pig ileum.

The type of antagonism exhibited by three novel bradykinin (BK) antagonists, D-Arg-[Hyp3,Thi5,D-Tic7,Oic8]BK (HOE 140, compound I), D-Arg-[Hyp3,D-Tic7,Oic8]BK (compound II) and [Arg(Tos)1,Hyp3,Thi5,D-Tic7,Oic8]BK (compound III), was compared with that of a conventional antagonist, D-Arg-[Hyp2,Thi5,8,D-Phe7]BK (compound IV), on the guinea-pig ileum. The novel compounds induced rightward displacements of cumulative concentration-response curves to BK, accompanied by a progressive reduction of the maximum effect (Emax) without a significant decrease in the slope, whereas no reduction of Emax was observed with compound IV. Actions of substance P on the guinea-pig ileum and of vasopressin on the rat uterus remained completely unaffected.

[Pain and stress: biological protection].

The biological methods of animal organism protection, the role of stress and the nociceptive signals, the hierarchy of defence mechanism--from the autonomic reflexes, neuroendocrine regulations, emotional reactions and expressions up to peculiar to the human being intellectual safeguard have been considered. The latest data on the role of tachykinins in transmission have been examined. The discovery of mediator function of "substance P" (SP), other related peptides, which coexists and realizes from the primary afferents and other terminals of SP-containing, capsaicin-sensitive neurons--opened a new chapter in the neurobiology and medicine.

Lack of significant unspecific effects of HOE 140 and other novel bradykinin antagonists in vitro and in vivo.

The novel, potent and long-acting bradykinin (BK) antagonists, HOE 140, compound II and compound III, slightly decreased blood pressure, but did not affect heart rate and respiration of rats. The antagonists did not cause bronchoconstriction in guinea-pigs. Neither HOE 140 nor BK released histamine from isolated perfused hindlegs of rats.