Monoclonal antibodies to dalargin--a synthetic analog of enkephalins.

Monoclonal antibodies (MABs) to a new drug Dalargin (Tyr-D-Ala-Gly-Phe-Leu-Arg) enhancing ulcer healing have been produced. Dalargin is a synthetic analog of Leu-enkephalin. With about 40 compounds tested in competition radioimmunoassay it has been shown that specificity of the MABs is directed against the N-terminal tetrapeptide of the molecule. The MABs are sensitive to amino acid substitutions in any of the positions of the fragment and have no cross-reactivity with endogenous opioids. Their further application in pharmacokinetic studies in humans and for characterization of opioid receptors is discussed.

Subcellular distribution of (+)-[3H]SKF 10,047 binding sites in rat liver.

The distribution of (+)-[3H]SKF 10,047 binding sites and the distribution of the established plasma membrane, nuclear, mitochondrial and endoplasmic reticulum markers in subcellular fractions of rat liver have been studied. The distribution profile of (+)-[3H]SKF 10,047 binding sites coincided with that of NADPH-cytochrome c reductase, the endoplasmic reticulum marker. (+)-[3H]SKF 10,047 binding sites in rat liver are therefore suggested to be located on the endoplasmic reticulum membrane and to represent a membrane-bound enzyme.

(+)-[3H]SK&F 10,047 binding sites in rat liver.

Stereospecific binding sites for (+)-[3H]SKF 10,047 have been demonstrated in rat liver. These binding sites were shown to be opioid receptors of the sigma type, and this was confirmed by their binding properties: reversibility, saturability, stereospecificity, effects of ions and various pharmacological drugs on (+)-[3H]SK sigma F 10,047 binding. The authors suggest that (+)-[3H]SKF 10,047 binding sites in liver and brain are a part of novel 'sigmergic' regulatory system.

[Isolation and characteristics of antibodies against synthetic fragments of oncoproteins. II. Antiserum against the transforming protein pp60src of Rous sarcoma virus]. / Poluchenie i kharakteristika antitel k sinteticheskim fragmentam onkobelkov. II. Antisyvorotka k transformiruiushchemu belku virusa sarkomy kur Rausa pp60src.

To generate the antibodies to the transforming protein of Rous sarcoma virus (RSV) pp60src, rabbits were immunized with the peptide, corresponding to 415-421 sequence of pp60src. These antibodies immunoprecipitate pp60src in RSV-transformed chicken and mammalian cells, and also some proteins (45, 85 and 120 kDa), which could be autophosphorylated in vitro. It was shown that 415-421 sequence of pp60src is not recognized by the antibodies to pp60src from RSV-induced tumour bearing rabbits (TBR serum). In contrast to TBR serum, antibodies, generated against synthetic peptide, corresponding 415-421 sequence of pp60src couldn't be phosphorylated in vitro, when [gamma-32P]ATP is added to the immune complex. The antipeptide antibodies, bound to pp60src did not block phosphorylation of TBR immunoglobulins, added to this immune complex. Hence, 415-421 sequence of pp60src RSV containing the major tyrosine phosphorylation site does not take part in the kinase reaction in vitro.

[Specific binding of N-allylnormetazocine (SKF 10047), a ligand of sigma-opioid receptors, with liver membranes]. / Spetsificheskoe sviazyvanie liganda sigma-opioidnykh retseptorov--N-allilnormetazotsina (SKF 10047)--s membranami pecheni.

A sigma-opioid receptor ligand, N-allylnormetazocine (SKF 10047), binds specifically and reversibly to rat liver membranes. The rat liver binding sites for SKF 10047 are similar to sigma-opioid CNS receptors. They fail to interact with classical opiates (morphine, naloxone) and opioid peptides but bind with high affinity benzomorphans (bremazocine, SKF 10047) and various psychotropic drugs (haloperidol, imipramine, phencyclidine etc).

[Study of peptide degradation in the serum by 1H-NMR]. / Issledovanie degradatsii peptidov v syvorotke krovi metodom 1H-IaMR.

Enzymatic degradation of hexapeptide Tyr-D-Ala-Gly-Phe-Leu-Arg in human serum has been investigated by 1H-NMR spectroscopy, and its pathways are suggested. Its final degradation products, tetra- and pentapeptide, are shown to be stable in human serum for several hours. The NMR spectroscopy application for pharmacokinetic studies is substantiated.

[Immunomorphological methods for identifying cells of the APUD system of the digestive organs]. / Immunomorfologicheskie metody identifikatsii kletok APUD-sistemy organov pishchevareniia.

Possibilities of application of immunomorphologic methods to gastrointestinal apud cells study were evaluated. These are: immunofluorescent, immunoenzyme, immune-peroxidase, peroxidase-antiperoxidase, avidinbiotin. Causes of immunologic and non-immunologic non-specific background staining and the means of its elimination are discussed, as well as problems of fixation and embedding of the material.

[Duodenal ulcer with gastrin cell hyperplasia--a special form of peptic ulcer]. / Iazva dvenadtsatiperstnoi kishki s giperplaznei gastrinovykh kletok--osoboia forma iazvennoi bolezni.

Pap test was used to study gastric and duodenal G and D cells, blood gastrin levels, basal and stimulated acid production, clinical manifestations, and morphological characteristics of the mucosa in 39 patients with duodenal ulcer and 13 controls. The findings enable the authors to outline a special form of peptic ulcer that is characterized by gastrin cellular hyperplasia concurrent with relative pyloric D cell deficit, gastric metaplasia in the duodenum and gastric hypersecretion. Such patients have more frequently multiple ulcers, predominantly 0(I) blood group, complication-aggravated course of the disease, and ulcer history in close relatives. Moreover, it has been demonstrated that incidence of gastric metaplasia is due to gastric hypersecretion; hyperplasia of duodenal gastrin cells is associated with incidence of gastric metaplasia in patients with peptic ulcer.

[Gastrointestinal endocrine cells in metaplasia of the gastric mucosa and duodenum]. / Gastrointestinal'nye éndokrinnye kletki pri metaplazii slizistoi obolochki zheludka i dvenadtsatiperstnoi kishki.

Grimelius reaction and immunohistochemical PAP method were used to study endocrine cells producing gastrin (G-cells), somatostatin (D-cells) and gamma-endorphin (GER-cells) in gastric and duodenal mucosa of 95 males with atrophic gastritis with intestinal and pyloric metaplasia. The number of cells was counted per 1 mm2 of the mucosa. In the cases of marked intestinal metaplasia the number of G-, GER- and especially D-cells in the pyloric region non-metaplastic epithelium decreases and is approaching to its number in the duodenum of the control group. In the foci of marked pyloric metaplasia of gastric corpus the number of G- and GER-cells is almost the same as in the zones of gastric metaplasia of duodenum, and is approximating their number in the pyloric region of controls, thus allowing the designation of pyloric metaplasia as a complete one.

[Gastric endocrine cells containing endorphin, gastrin and somatostatin in duodenal ulcer]. / Endokrinnye kletki zheludka, soderzhashchie endorfin, gastrin i somatostatin, pri iazvennoi bolezni dvenadtsatiperstnoi.

Three types of endocrine cells (G cells producing gastrin-17, D cells producing somatostatin, and GER cells containing endorphine) in the mucous membrane of the stomach antrum from 14 patients with duodenal ulcer and 10 healthy persons were studied. Biopsies were fixed in a modified Bowen solution and imbedded into paraffin. The slides were stained by Grimelins' method and immunohistochemically with the use of the peroxidase-antiperoxidase method. The number of cells per 1 mm2 of the mucous membrane was counted. Patients with ulcer have shown the increased number of G and GER cells and decreased number of D cells. Besides, pronounced G cell hyperplasia with a relative decrease of GER cells and a marked decrease of Grimelins-positive cells (as compared to other patients with duodenal ulcer) were observed in 3 out of 14 ulcer patients. The authors conclude that the alteration of the balance between antagonistic hormone effects results in the hypersecretory syndrome that plays the main role in the pathogenesis of duodenal ulcer.

[Pancreatic endocrine function in persons with a history of brucellosis]. / Endokrinnaia funktsiia podzheludochnoi zhelezy u lits, perenesshikh brutsellez.

Different food tests were used to study endocrine function of the pancreas in 75 persons with a history of brucellosis and in 30 patients with chronic acalculous cholecystitis. In persons with a history of brucellosis, the endocrine part of the pancreas was involved, which manifested itself in a decrease in the secretory response of B cells to stimulation with glucose and mixed food enriched with proteins and fat. The alterations described are likely to be caused by the infection itself.

[Dynamics of the basal gastrin concentration of duodenal ulcer patients during the development of remission of the disease]. / Dinamika bazal'noi kontsentratsii gastrina u bol'nykh duodenal'noi iazvoi v protsesse stanovleniia remissii zabolevaniia.

The authors describe the results of studying basal gastrin secretion in patients with peptic ulcer of the duodenum at different phases of disease treated in 3 stages. It was revealed that basal secretion of gastrin experienced substantial changes in the course of transition from the phase of exacerbation to the phase of disease remission. It was noted that the onset of the clinico-endoscopic remission of peptic ulcer did not correlate in all the cases with normalization of basal gastrin level. Patients who did not show normalization of basal gastrin level during treatment were more prone to the development of repeated exacerbations. Based on the data obtained the authors determined the prognostic importance of radioimmunoassay of gastrin basal concentration over time. The increment of gastrin concentration by more than 35 pg/ml of the level seen during peptic ulcer exacerbation is a prognostically unfavourable sign, for the probability of relapses rises up to 79% during a year.

[A synthetic peptide from Helix aspersa raises arterial pressure in rats]. / Sinteticheskii peptid iz Helix aspersa povyshaet arterial'noe davlenie u krys.

New FMRF-amide like peptide, pGlu-Asp-Pro-Phe-Leu-Arg-Phe-NH2, originally isolated from the ganglia of Helix aspersa, was synthetized. Intravenous injections of this peptide (40-300 micrograms/kg) produce rapid dose dependent increase in the blood pressure and heart rate of anaesthetised rats. Since alpha-adrenoreceptor blockade with prazosin eliminated pressor response, it is concluded that the increase in the arterial pressure is mediated by sympathetic activation.

[Clinical evaluation of hexapeptide dalargine used in the treatment of duodenal ulcer]. / Klinicheskaia otsenka geksapeptida dalargina kak sredstva lecheniia iazvennoi bolezni dvenadtsatiperstnoi kishki.

Experiments on rats were made to study antiulcerous activity of the majority of endogenous opioid-like peptides and 22 synthetic analogs. Duodenal ulcers were induced in animals by cysteamine hydrochloride. A hexapeptide called dalargin demonstrated the maximal antiulcerous activity in experimental duodenal ulcer. The drug dose applied was 10-15 micrograms/kg. The drug action was found to be mediated via opiate receptors. The drug did not cause the development of addiction or physical dependence. Dalargin was studied in 45 male patients with exacerbation of duodenal ulcer. The mean period of healing amounted to 21.8 days. The use of dalargin in a dose of 2 mg a day intramuscularly produced no side effects or changes in blood characteristics. The mechanisms of dalargin action are discussed from the viewpoint of the general principles of peptide pharmacology.

[The dynamics of the peptide hydra morphogen in meningitis]. / Dinamika peptidnogo morfogena gidry pri meningitakh.

The content of peptide hydra morphogen (PHM) was measured for the first time in blood plasma and CSF by RIA in patients with purulent meningitides. 36 patients were examined. Blood and CSF were withdrawn in the acute period and during early convalescence. In the reference group (patients with acute respiratory diseases of virus etiology, quinsy, pneumonias), the PHM concentration in the blood was 128 +/- 62 fm/ml, that in CSF 66 +/- 14 fm/ml. In purulent meningitis of medium gravity, the content of PHM in blood plasma and CSF did not significantly differ from the control. Grave purulent meningitis was marked by a high content of PHM in blood plasma and CSF, in particular, in the acute period and during early convalescence. It was 1250.30 +/- 191 and 544.03 +/- 159 fm/ml in blood plasma and 1504.38 +/- 475 and 1811.39 +/- 375 fm/ml in CSF, respectively. In grave complicated purulent meningitis, which was mainly observed in persons over 50 years, the content of PHM in blood plasma exceeded but insignificantly the content in the control during the acute period (327.49 +/- 76 fm/ml), followed by further increase (1131.41 +/- 209 fm/ml) by days 8-12 of the disease; within the same time, the content of PHM in CSF did not significantly differ from the control (319.52 +/- 152 and 625.69 +/- 275 fm/ml, respectively). In view of the fact that hypothalamic PHM is of crucial importance in the processes of nervous tissue regeneration, the content of the latter one in CSF in patients with grave purulent meningitis may serve as a criterion for disease course tendencies.

[Receptor characteristics of the duodenal mucosa in peptic ulcer: a new approach to the individual selection of the therapy]. / Retseptornaia kharakteristika duodenal'noi slizistoi pri iazvennoi bolezni: novyi podkhod k individualizirovannomu vyboru terapii.

The radioligand-receptor method was used to assay the M-cholinereceptors and opioid binding centers in biopsy specimens of the duodenal mucosa from 40 patients with duodenal ulcer as compared with the results of subsequent therapy by means of gastrocepin (20 patients) and dalargin (20 patients). Analysis of the isothermal curve of 3H-QNB-binding with duodenal mucosa membranes within Skatchard's coordinates showed the presence of 2 subtypes of muscarinic receptors blocked (M1) and not blocked (M2) by gastrocepin. The majority of the patients in both groups demonstrated a significant decrease of the dissociation constant as compared to the control (20 patients). In patients with high receptor sensitivity to gastrocepin and dalargin, ulcer cicatrization was noted during the treatment in all the cases. Meanwhile the treatment proved little effective in patients with appreciably decreased receptor sensitivity to the drugs under consideration. The data obtained indicate the possibility of predicting the results of duodenal ulcer treatment by means of the radioreceptor analysis, providing the basis for the individualized treatment choice.

[Gastrin, gamma-endorphin and somatostatin cells of the stomach and duodenum in patients with duodenal ulcer treated with dalargin]. / Gastrinovye, gamma-éndorfinovye i somatostatinovye kletki zheludka i dvenadtsatiperstnoi kishki u bol'nykh iazvennoi bolezn'iu dvenadtsatiperstnoi kishki pri lechenii dalarginom.

The distribution of gastrin-, gamma-endorphin and somatostatin-producing cells in antral and duodenal mucosa was studied in biopsies from 26 patients with duodenal ulcer and from 13 controls by the immunohistochemical (PAP) method. The number of antral somatostatin-producing cells after dalargin treatment was significantly higher in comparison with controls and patients treated with antacids plus atropine. These changes may be connected with the antiulcer activity of dalargin, a new opioid peptide drug.

[The synthetic peptide preparation dalargin in the treatment of peptic ulcer]. / Sinteticheskii peptidnyi preparat dalargin v lechenii iazvennoi bolezni.

The paper is concerned with the results of clinical assessment of a new peptide drug dalargin in therapy of duodenal peptic ulcer. Assessment of the efficacy of dalargin was compared to that of tagamet and placebo using double "blind" control. A total of 180 males with exacerbation of duodenal ulcer were examined, of them 90 received dalargin only, 70 received tagamet and 20 persons participated in an experiment based on a method of double "blind" control. An average period of duodenal ulcer healing in the patients on dalargin therapy was 21.4 +/- 1.2 days, and by the 28th day complete cicatrization was observed in 87.5% of the patients. In tagamet therapy an average period of cicatrization was 23.7 +/- 1.8 days, by the 28th day ulcer healing was observed in 87.3%. With placebo ulcer healing by the 4th week was observed in 30% of the examinees, in dalargin therapy in 80%. A conclusion was made of high efficacy of dalargin for therapy of peptic ulcer.