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PubMed1988

Fatal intoxications in Denmark following intake of morphine from opium poppies.

In Denmark it is legal to grow opium poppies for the production of poppy seeds and until 1986 for decoration purposes, too. Danish poppy capsules contain 0.3-5 mg morphine per capsule and the content of morphine in opium exuded from the capsules may amount to 24%. This has resulted in misuse as both fresh and dried poppy capsules have been used for the production of "opium tea". During the period 1982-1985, seven casualties occurred among drug addicts in Denmark which were solely or partly caused by these opium poppies.

Adolescent↗
PubMed1980

Rapid method for extraction and colorimetric estimation of morphine in Indian opium.

A rapid method for the extraction and estimation of morphine in Indian opium is described. Methanolic extract of opium is treated with activated charcoal and the colorless extract obtained contains all of the major opium alkaloids. Morphine content is estimated colorimetrically by using hydrogen peroxide-copper-ammonia. The reaction is specific for morphine, and no other major opium alkaloid responds. The results obtained by the proposed method are in general agreement with those obtained by the nitroso method.

Colorimetry↗
PubMed1995

An ethnographic account of opium consumers of Rajasthan (India): socio-medical perspective.

This communication gives an ethnographic account of 200 opium users in selected villages of three Western districts in the desert state of Rajasthan. The region is known for its traditional use of raw opium in the form of amal or doda, due to its climate and difficult living conditions. The ethnographic information suggests that opium use is in many ways integrated into the socio-cultural fabric of the local community. Self-medication with opium mitigates various health problems and the drug is also used to relieve mental distress. Besides these uses for relief of distress, the drug is used recreationally and within settings which facilitate social bonding. The traditional roles ascribed to the use of this intoxicant cannot be dismissed when formulating long-term preventive and control measures.

Adolescent↗
PubMed1994

Fifty years of development of opium characterization methods.

In view of the recent call by the Sub-Commission on Illicit Drug Traffic and Related Matters in the Near and Middle East, for the "development of mechanisms to identify, with more precision and through laboratory analysis, the sources of opium seized from the illicit traffic" [1], the present paper reviews the rationale and preconditions for any practical and reliable characterization and origin-correlated classification of opium. In that context, the results of the early international efforts under the aegis of the United Nations from 1951 to 1967, as well as the rather sporadic investigations in this direction since 1968, are described. Finally, it is demonstrated that in spite of the application of modern computer-based technology, the main obstacle to comprehensive opium characterization and typology is still the lack of an extensive reference collection of opium samples of known origin.

Chemistry Techniques, Analytical↗
PubMed2003

Developmental and inducible accumulation of gene transcripts involved in alkaloid biosynthesis in opium poppy.

Opium poppy (Papaver somniferum) produces a large number of benzylisoquinoline alkaloids, including morphine and sanguinarine, derived from tyrosine via the branch-point intermediate (S)-reticuline. Molecular clones for the three methlytransferases involved in (S)-reticuline biosynthesis, (S)-norcoclaurine-6-O-methyltransferase (6OMT), (S)-3'-hydroxy-N-methylcoclaurine-4'-O-methyltransferase (4'OMT), and (S)-coclaurine N-methyltransferase (CNMT), were isolated from opium poppy and shown to share extensive homology with the corresponding cDNAs from Japanese goldthread (Coptis japonica). These cDNAs were used together with previously isolated clones for tyrosine/dopa decarboxylase (TYDC), (S)-N-methylcoclaurine-3'-hydroxylase (CYP80B1), berberine bridge enzyme, (BBE), (7S)-salutaridinol 7-O-acetyltransferase (SAT), and codeinone reductase (COR), to compare the accumulation of gene transcripts encoding eight alkaloid biosynthetic enzymes in opium poppy. Transcript levels generally increased in developing seedlings and were consistently high in stems and flower buds, but were more variable in roots and leaves of mature plants. The accumulation of each transcript, with the exception of COR, showed a marked induction in response to elicitor treatment or wounding of cultured cells. Specific gene transcript levels often correlated with the accumulation of morphine or sanguinarine, with notable exceptions. Our data suggest some degree of coordination in the developmental and inducible regulation of alkaloid biosynthetic genes in opium poppy.

Alkaloids↗
PubMed2002

Buprenorphine treatment of opium-dependent outpatients seeking treatment in Iran.

The goal of this study was to evaluate the efficacy of 1-, 3-, and 8-mg per day doses of buprenorphine in the maintenance treatment of opium-dependent patients over a 6-month treatment period. Participants were 513 opium-dependent individuals who were seeking treatment in an urban outpatient clinic, offering a 1-hr weekly individual counseling session. Overall, 305 patients (59.5%) completed the 6-month study. Completion rates by dosage group were 33.9% for the 1-mg dose group, 64.3% for the 3-mg dose group, and 80.1% for the 8-mg dose group-each significantly different from the other two groups. The results support the efficacy and safety of buprenorphine for outpatient treatment of opium dependence and seem to indicate that the highest dose (8 mg) of buprenorphine was the best of the three doses for Iranian opium-dependent patients to increase their retention in treatment.

Adult↗
PubMed1999

Analysis of promoters from tyrosine/dihydroxyphenylalanine decarboxylase and berberine bridge enzyme genes involved in benzylisoquinoline alkaloid biosynthesis in opium poppy.

Tyrosine/dihydroxyphenylalanine decarboxylase (TYDC) and the berberine bridge enzyme (BBE) represent the entry point and a key branch point, respectively, in the biosynthesis of benzylisoquinoline alkaloids in select species of the Papaveraceae and Fumariaceae. Genomic clones for tydc7 and bbe1 from opium poppy (Papaver somniferum L.) were isolated. Deletion analysis of tydc7 and bbe1 5'-flanking regions revealed the location of putative regulatory domains necessary for expression of the beta-glucuronidase (gus) reporter gene in a transient assay system based on the microprojectile bombardment of cultured opium poppy cells. A 105-nucleotide region between -393 and -287 of the tydc7 5'-flanking region, and a 155-nucleotide region between -355 and -200 of the bbe1 5'-flanking region, were found to be essential for promoter activity. RNA gel blot analysis showed that tydc7 and bbe1 expression is induced in cultured opium poppy cells in response to wounding or treatment with a pathogen-derived elicitor. Time-courses for the induction of tydc7 and bbe1 mRNAs in wounded cells were nearly identical to those for GUS activity in cells bombarded with select promoter-gus constructs when the -393 to -287 region of tydc7, or the -355 to -200 region of bbe1, was present. Our data suggest that the wound signal caused by the entry of DNA-coated microcarriers into opium poppy cells was sufficient to induce tydc7 and bbe1 promoter activity, and that wound-responsive regulatory elements are located within domains identified by deletion analysis.

Alkaloids↗
PubMed2001

Morphine metabolism in the opium poppy and its possible physiological function. Biochemical characterization of the morphine metabolite, bismorphine.

We identified a novel metabolic system of morphine in the opium poppy (Papaver somniferum L.). In response to stress, morphine is quickly metabolized to bismorphine consisting of two morphine units, followed by accumulation in the cell wall. This bismorphine binds predominantly to pectins, which possess high galacturonic acid residue contents, through ionical bonds. Our newly developed method using artificial polysaccharides demonstrated that bismorphine bridges are formed between the two amino groups of bismorphine and the carboxyl groups of galacturonic acid residues, resulting in cross-linking of galacturonic acid-containing polysaccharides to each other. The ability of bismorphine to cross-link pectins is much higher than that of Ca2+, which also acts as a cross-linker of these polysaccharides. Furthermore, we confirmed that cross-linking of pectins through bismorphine bridges leads to resistance against hydrolysis by pectinases. These results indicated that production of bismorphine is a defense response of the opium poppy. Bismorphine formation is catalyzed by anionic peroxidase that pre-exists in the capsules and leaves of opium poppies. The constitutive presence of morphine, together with bismorphine-forming peroxidase, enables the opium poppy to rapidly induce the defense system.

Carbohydrate Sequence↗
PubMed2000

Agrobacterium rhizogenes-mediated transformation of opium poppy, Papaver somniferum l., and California poppy, Eschscholzia californica cham., root cultures.

An efficient protocol for the establishment of transgenic opium poppy (Papaver somniferum L.) and California poppy (Eschscholzia californica Cham.) root cultures using A. grobacterium rhizogenes is reported. Five strains of A. rhizogenes were tested for their ability to produce hairy roots on wounded opium poppy seedlings and California poppy embryogenic calli. Three of the strains induced hairy root formation on both species, whereas two others either caused the growth of tumorigenic calli or produced no response. To characterize the putative transgenic roots further, explant tissues were co-cultivated with the most effective A: rhizogenes strain (R1000) carrying the pBI121 binary vector. Except for the co-cultivation medium, all formulations included 50 mg l(-1) paromomycin to select for transformants and 200 mg l(-1) timentin to eliminate the Agrobacterium. Four weeks after infection, paromomycin-resistant roots appeared on 92-98% of explants maintained on hormone-free medium. Isolated hairy roots were propagated in liquid medium containing 1.0 mg l(-1) indole-3-acetic acid to promote rapid growth. Detection of the neomycin phosphotransferase gene, high levels of beta-glucuronidase (GUS) transcripts and enzyme activity, and GUS histochemical localization confirmed the integrative transformation of root cultures. Transgenic roots grew faster than wild-type roots, and California poppy roots grew more rapidly than those of opium poppy. With the exception of a less compact arrangement of epidermal cells and more root hairs, transformed roots of both species displayed anatomical features and benzylisoquinoline alkaloid profiles that were virtually identical to those of wild-type roots. Transgenic root cultures of opium poppy and California poppy are a simple, reliable and well-defined model system to investigate the molecular and metabolic regulation of benzylisoquinoline alkaloid biosynthesis, and to evaluate the genetic engineering potential of these important medicinal plants.

Alkaloids↗
PubMed2005

Sanguinarine biosynthesis is associated with the endoplasmic reticulum in cultured opium poppy cells after elicitor treatment.

Three key benzylisoquinoline alkaloid biosynthetic enzymes, (S)-N-methylcoclaurine-3'-hydroxylase (CYP80B1), berberine bridge enzyme (BBE), and codeinone reductase (COR), were localized in cultured opium poppy (Papaver somniferum) cells by sucrose density gradient fractionation and immunogold labeling. CYP80B1 catalyzes the second to last step in the formation of (S)-reticuline, the last common intermediate in sanguinarine and morphine biosynthesis. BBE converts (S)-reticuline to (S)-scoulerine as the first committed step in sanguinarine biosynthesis, and COR catalyzes the penultimate step in the branch pathway leading to morphine. Sanguinarine is an antimicrobial alkaloid that accumulates in the vacuoles of cultured opium poppy cells in response to elicitor treatment, whereas the narcotic analgesic morphine, which is abundant in opium poppy plants, is not produced in cultured cells. CYP80B1 and BBE were rapidly induced to high levels in response to elicitor treatment. By contrast, COR levels were constitutive in the cell cultures, but remained low and were not induced by addition of the elicitor. Western blots performed on protein homogenates from elicitor-treated cells fractionated on a sucrose density gradient showed the cosedimentation of CYP80B1, BBE, and sanguinarine with calreticulin, and COR with glutathione S-transferase. Calreticulin and glutathione S-transferase are markers for the endoplasmic reticulum (ER) and the cytosol, respectively. In response to elicitor treatment, large dilated vesicles rapidly developed from the lamellar ER of control cells and fused with the central vacuole. Immunogold localization supported the association of CYP80B1 and BBE with ER vesicles, and COR with the cytosol in elicitor-treated cells. Our results show that benzylisoquinoline biosynthesis and transport to the vacuole are associated with the ER, which undergoes major ultrastructural modification in response to the elicitor treatment of cultured opium poppy cells.

Alcohol Oxidoreductases↗
PubMed1996

Molecular analysis of a new member of the opium poppy tyrosine/3,4-dihydroxyphenylalanine decarboxylase gene family.

An aromatic amino acid decarboxylase DNA fragment was generated from opium poppy (Papaver somniferum L.) genomic DNA by the PCR using primers designed from conserved amino acid sequences of other aromatic amino acid decarboxylase genes. Using this fragment as a probe, a genomic clone was isolated that encodes a new member of the opium poppy tyrosine/3,4-dihydroxyphenylalanine decarboxylase gene family (TyDC5). The predicted TyDC5 amino acid sequence shares extensive identity with other opium poppy tyrosine/3,4-dihydroxyphenylalanine decarboxylases (84%), and when expressed in Escherichia coli, it is active against tyrosine and to a lesser extent against 3,4-dihydroxyphenylalanine. Ribonuclease protection assays indicate that TyDC5 is expressed primarily in the roots of mature poppy plants. A peak of TyDC5 expression was also observed during germination, coincident with the emergence of the radicle from the seed coat. Parallel results were obtained in transgenic tobacco using a TyDC5 promoter fragment (-2060) translationally fused to the beta-glucuronidase reporter gene (GUS). IN TyDC5::GUS tobacco, GUS activity transiently appeared in all parts of the seedling during germination, but was limited to the roots in older plants. These results indicate that TyDC5 expression is transcriptionally regulated and suggest that the TyDC5 enzyme may play an important role in providing precursors for alkaloid synthesis in the roots and germinating seedlings of opium poppy.

Amino Acid Sequence↗
PubMed1998

Expression patterns conferred by tyrosine/dihydroxyphenylalanine decarboxylase promoters from opium poppy are conserved in transgenic tobacco.

Opium poppy (Papaver somniferum) contains a large family of tyrosine/dihydroxyphenylalanine decarboxylase (tydc) genes involved in the biosynthesis of benzylisoquinoline alkaloids and cell wall-bound hydroxycinnamic acid amides. Eight members from two distinct gene subfamilies have been isolated, tydc1, tydc4, tydc6, tydc8, and tydc9 in one group and tydc2, tydc3, and tydc7 in the other. The tydc8 and tydc9 genes were located 3.2 kb apart on one genomic clone, suggesting that the family is clustered. Transcripts for most tydc genes were detected only in roots. Only tydc2 and tydc7 revealed expression in both roots and shoots, and TYDC3 mRNAs were the only specific transcripts detected in seedlings. TYDC1, TYDC8, and TYDC9 mRNAs, which occurred in roots, were not detected in elicitor-treated opium poppy cultures. Expression of tydc4, which contains a premature termination codon, was not detected under any conditions. Five tydc promoters were fused to the beta-glucuronidase (GUS) reporter gene in a binary vector. All constructs produced transient GUS activity in microprojectile-bombarded opium poppy and tobacco (Nicotiana tabacum) cell cultures. The organ- and tissue-specific expression pattern of tydc promoter-GUS fusions in transgenic tobacco was generally parallel to that of corresponding tydc genes in opium poppy. GUS expression was most abundant in the internal phloem of shoot organs and in the stele of roots. Select tydc promoter-GUS fusions were also wound induced in transgenic tobacco, suggesting that the basic mechanisms of developmental and inducible tydc regulation are conserved across plant species.

Amino Acid Sequence↗
PubMed2006

The role of phloem sieve elements and laticifers in the biosynthesis and accumulation of alkaloids in opium poppy.

The benzylisoquinoline alkaloids of opium poppy, including the narcotic analgesics morphine and codeine, accumulate in the multinucleate cytoplasm of specialized laticifers that accompany vascular tissues throughout the plant. In mature opium poppy plants, immunofluorescence labeling using specific antibodies showed that four alkaloid biosynthetic enzymes, (S)-norcoclaurine 6-O-methyltransferase (6OMT), (S)-coclaurine N-methyltransferase (CNMT), (S)-3'-hydroxy-N-methylcoclaurine-4'-O-methyltransferase (4'OMT) and salutaridinol-7-O-acetyltransferase (SAT) were restricted to sieve elements of the phloem adjacent or proximal to laticifers. The identity of sieve elements was confirmed by (i) the specific immunogold labeling of the characteristic cytoplasm of this cell type, (ii) the co-localization of a sieve element-specific H(+)-ATPase with all biosynthetic enzymes and (iii) the strict association of sieve plates with immunofluorescent cells. The localization of laticifers was demonstrated antibodies specific to major latex protein (MLP), which is characteristic of this cell type. In situ hybridization using antisense RNA probes for 6OMT, CNMT, 4'OMT and SAT showed that the corresponding gene transcripts were found in the companion cell paired with each sieve element. Seven benzylisoquinoline alkaloid biosynthetic enzymes, (S)-N-methylcoclaurine 3'-hydroxylase (CYP80B1), berberine bridge enzyme, codeinone reductase, 6OMT, CNMT, 4'OMT and SAT were localized by immunofluorescence labeling to the sieve elements in the root and hypocotyl of opium poppy seedlings. The abundance of these enzymes increased rapidly between 1 and 3 days after seed germination. The localization of seven biosynthetic enzymes to the sieve elements provides strong support for the unique, cell type-specific biosynthesis of benzylisoquinoline alkaloids in the opium poppy.

Alkaloids↗
PubMed2000

[Alcohol abuse by patients with opium addiction].

A problem of alcohol abuse in patients with opium addiction was investigated. 72 patients with were examined. An alcoholic positive family history was investigated in families of the patients as well as frequency and character of premorbid alcohol abuse, types of alcoholization in remission of opium addiction and relationship between alcohol drinking in drug addiction and psychopathologic disorders. A correlation between massive alcoholization and worsening of the somatic state of the patients was determined. It was found that opium addiction increases predisposition to alcohol abuse. Relationship between massive alcoholization and unfavorable course of opium addiction is shown.

Adolescent↗
PubMed2002

Advanced treatment of high strength opium alkaloid industry effluents.

The purpose of this study was to investigate an effective treatment system which can be applicable to treat opium alkaloid industry (OAI) effluents characterised with high COD, TKN, dark color and non-biodegradable organic pollutants. In the first phase of the study, lab-scale anaerobic (UASBR) + aerobic (SBR) treatability studies were carried out on opium processing industry effluents. Effluent CODs from the two staged biological treatment system were relatively high (-700 mg l(-1)) and additional post treatment was required. Physico-chemical treatability studies previously carried out on the effluent of opium alkaloid wastewater treatment plant, were not effective in removing residual COD and color. In the second phase of the study, the refractory organics causing higher inert COD values in the SBR effluent were additionally treated by using Fenton's Oxidation. The batch tests were performed to determine the optimum operating conditions including pH, H2O2 dosage, molar ratio of Fe2+/H2O2 and reaction time. It was found that removal efficiencies of COD and color for 30 minutes reaction time were about 90% and 95%, respectively. The ratio of H2O2/FeSO4 was determined as 200 mg l(-1)/600 mg l(-1) for the optimum oxidation and coagulation process at pH 4. Experimental results of the present study have clearly indicated that the Fenton's oxidation technology is capable to treat almost all parts of the organics which consist of both soluble initial and microbial inert fractions of COD for opium alkaloid industry effluents. Effluents from the Fenton's Oxidation process can satisfy effluent standards for COD and color in general.

Bacteria, Aerobic↗
PubMed1976

The pro-heroin effects of anti-opium laws in Asia.

Over 25 years anti-opium laws were enacted by three Asian governments in countries where opium use was traditional. Within months, heroin use suddenly appeared; and within a decade, heroin addiction surpassed opium addiction. The laws led to (1) increased price of narcotic drugs, (2) a heroin "industry," (3) corruption of the law enforcement system, and (4) major health problems involving parenteral drug use. The Asian experience indicates that antinarcotic laws can be effective only with careful preparations: (1) changing society's attitude toward the traditional drug from ambivalence to opposition; (2) mobilizing resources to treat and rehabilitate all addicts within a short period of time; (3) developing the social will to incarcerate all "recidivist" addicts for a prolonged period; and (4) preventing narcotic production or importation.

Drug and Narcotic Control↗
PubMed2004

Rapid and sensitive determination of morphine in street opium samples by thermal desorption gas chromatography using a microfurnace pyrolyzer.

Thermal desorption of the alkaloids in opium samples at 300 degrees C using a vertical microfurnace pyrolyzer was followed by their on-line gas chromatographic (GC) analysis on a large-bore glass capillary column. This method permitted rapid and sensitive determination of the content of the main alkaloid, morphine, in the small (ca. 100 microg) opium samples with a relative standard deviation within 4% for 5 runs. The observed morphine contents of about 12 to 15 w/w% in the given opium samples were in fairly good agreement with those estimated by a conventional GC-MS method.

Alkaloids↗
PubMed2005

The GC-MS detection and characterization of neopine resulting from opium use and codeine metabolism and its potential as an opiate-product-use marker.

Neopine, a minor opium alkaloid and an isomer of codeine (also known as beta-codeine), has been detected in both the urine of opium users and pharmaceutical codeine users. The characterization of neopine was achieved by comparison of the mass spectra and GC retention times of the trimethylsilyl derivative. The presence of neopine in the urine of pharmaceutical codeine users was attributed to the metabolism of codeine through a double bond migration in ring C, from the 7-8 to the 8-14 position. The potential use of the alkaloid as a confirmation marker of opium and/or pharmaceutical codeine use and the ability to differentiate these from heroin use has been discussed.

Biomarkers↗