The tissue distribution, course of secretion, and sex differences of morphine were delineated in ... more The tissue distribution, course of secretion, and sex differences of morphine were delineated in Ascaris suum. Nitric oxide (NO) release in various tissues in response to morphine and its metabolite morphine-6-glucuronide (M6G) were also examined. Ascaris suum of both sexes along with their incubation fluid were analyzed for morphine concentrations by high- performance liquid chromatography (HPLC) over a 5-day period. Various tissues were also dissected for HPLC and NO analysis. Morphine was found to be most prevalent in the muscle tissue, and there is significantly more morphine in females than males, probably because of the large amounts present in the female uterus. Morphine (10−9 M) and M6G (10−9 M) stimulated the release of NO from muscles. Naloxone (10−7 M) and N-nitro-l-arginine methyl ester (10−6 M) blocked (P < 0.005) morphine-stimulated NO release from A. suum muscle tissue. d-Phe-Cys-Tyr-d-Trp-Om-Thr-Pen-Thr-NH2 (CTOP) (10−7 M) did not block morphine's NO release. ...
The effects of the opiates morphine and morphine-6-glucuronide (M6G), the μ opioid receptor speci... more The effects of the opiates morphine and morphine-6-glucuronide (M6G), the μ opioid receptor specific antagonist D-Phe-Cys-Tyr-D-Trp-Om-Thr-Pen-Thr-NH 2 (CTOP), and the general opiate antagonist naloxone on the latency of response to thermal stimulation were determined in the parasitic nematode Ascaris suum. Thermal detection and avoidance behaviors of the worms were evaluated with a tail flick analgesia meter using a modification of a technique employed for nociception experiments in rodents. Morphine and M6G were shown to have a dose dependent analgesic effect on A. suum's latency of response to heat with morphine being the most potent. The analgesic effect of morphine was reversed by naloxone but not CTOP. Neither naloxone nor CTOP was able to block the analgesia of M6G. CTOP but not naloxone had significant analgesic effects on its own. These findings are generally consistent with previous results on the effects of opiates and nitric oxide release from A. suum tissue. Apparently these nematodes possess opioid receptors that effect nociception.
Samples of mosquitoes in the Culex pipiens L. complex from Memphis, Tenn., were collected from Ju... more Samples of mosquitoes in the Culex pipiens L. complex from Memphis, Tenn., were collected from June to November 1985 and examined in regard to allozyme frequencies and ratios of the two arms of the phallosome of the male genitalia (DV/D). The dominant allozymes of hexokinase (HkA) and 6-phosphogluconate dehydrogenase (PgdF) significantly increased in frequency during this period as did the mean DV/D ratio. An analysis of gene frequencies by species group designated by DV/D ratios revealed no significant differences among Cx. pipiens, Cx. quinquefasciatus Say, and intermediates. The lack of association of gene frequencies with the taxa determined by the DV/D ratio indicated that although allozyme frequencies were correlated temporally with the DV/D ratio in the population, they were not associated with subspecies. These results are consistent with previous work that has shown latitudinal association and thermal stability differences in the major allozymes of these enzymes in the Cx. ...
Medical science monitor : international medical journal of experimental and clinical research, 2005
The current paper summarizes the findings on endogenous morphine isolated by HPLC and characteriz... more The current paper summarizes the findings on endogenous morphine isolated by HPLC and characterized by mass spectroscopy in Schistosoma mansoni, Dracunculus medinensis and Ascaris suum. Morphine-6-glucuronide (M6G) has also been found by HPLC and confirmed by mass spectroscopy in Dracunculus medinensis and Ascaris suum. In addition, a morphine like substance has been isolated from Trichinella spiralis by HPLC and mice infected with Trichinella spiralis show a naloxone reversible analgesia. We discuss in greater detail the tissue distribution, course of secretion, and sex differences of morphine in Ascaris suum. Finally, we explore the function of morphine as both an internal signaling molecule and its use in immune evasion in Ascaris suum.
The effects of the opiates morphine and morphine-6-glucuronide (M6G), the mu opioid receptor spec... more The effects of the opiates morphine and morphine-6-glucuronide (M6G), the mu opioid receptor specific antagonist D-Phe-Cys-Tyr-D-Trp-Om-Thr-Pen-Thr-NH(2) (CTOP), and the general opiate antagonist naloxone on the latency of response to thermal stimulation were determined in the parasitic nematode Ascaris suum. Thermal detection and avoidance behaviors of the worms were evaluated with a tail flick analgesia meter using a modification of a technique employed for nociception experiments in rodents. Morphine and M6G were shown to have a dose dependent analgesic effect on A. suum's latency of response to heat with morphine being the most potent. The analgesic effect of morphine was reversed by naloxone but not CTOP. Neither naloxone nor CTOP was able to block the analgesia of M6G. CTOP but not naloxone had significant analgesic effects on its own. These findings are generally consistent with previous results on the effects of opiates and nitric oxide release from A. suum tissue. Appar...
R ECENT studies have used the blue mussel, Mytilus edulis, as a sentinel organism for pollution (... more R ECENT studies have used the blue mussel, Mytilus edulis, as a sentinel organism for pollution (Greig & Sennefelder 1985; Micallef & Tyler 1989). The mussel and other marine mollusks, such as oysters and clams, have long been known to concentrate toxic heavy metals, such as mercury, lead and cadmium. They are also capable of bioaccumulating synthetic organic compounds, such as PCBs and pesticides. This propensity to concentrate toxins often poses a public health problem, but it also offers an opportunity for environmental monitoring. The United Nations Environmental Program has in fact launched a global effort to monitor these organisms for marine pollution (Bayne 1989; Beeby 1993; Borchart et al. 1988). Not only can mussels act as reservoirs to measure the extent of chemical contamination and its variation in different coastal environments, it can also be used in the teaching lab to illustrate biological responses to pollution (Bayne 1989; Conrad 1988). Pollutants, such as heavy metals, have been found to alter molecular and cellular processes in mussels. While each organism is unique in both its capacity to deal with pollutants and its biological responses to them, certain cellular responses are often similar down the phylogenetic scale. A serious concern in recent years is the immune suppressive effect that pollutants are having on marine life. This phenomenon has been observed from invertebrate organisms, such as mussels and oysters, to vertebrates such as fish, seals and whales (De L. Swart et al. 1994; Faisal et al. 1991; Sami et al. 1992). The blue mussel offers an easy way of introducing your class to both immune response and the effects of environmental pollution on marine organisms. The immune cells of mollusks are called hemocytes and can be easily extracted with hemolymph from the mussel's foot or heart with a syringe. These cells have been shown to have many characteristics of vertebrate granular leukocytes. They respond to the same mes-
The tissue distribution, course of secretion, and sex differences of morphine were delineated in ... more The tissue distribution, course of secretion, and sex differences of morphine were delineated in Ascaris suum. Nitric oxide (NO) release in various tissues in response to morphine and its metabolite morphine-6-glucuronide (M6G) were also examined. Ascaris suum of both sexes along with their incubation fluid were analyzed for morphine concentrations by highperformance liquid chromatography (HPLC) over a 5-day period. Various tissues were also dissected for HPLC and NO analysis. Morphine was found to be most prevalent in the muscle tissue, and there is significantly more morphine in females than males, probably because of the large amounts present in the female uterus. Morphine (10 Ϫ9 M) and M6G (10 Ϫ9 M) stimulated the release of NO from muscles. Naloxone (10 Ϫ7 M) and N-nitro-L-arginine methyl ester (10 Ϫ6 M) blocked (P Ͻ 0.005) morphine-stimulated NO release from A. suum muscle tissue. D-Phe-Cys-Tyr-D-Trp-Om-Thr-Pen-Thr-NH 2 (CTOP) (10 Ϫ7 M) did not block morphine's NO release. However, naloxone could not block M6G-stimulated NO release by muscles, whereas CTOP (10 Ϫ7 M) blocked its release. These findings were in seeming contradiction to our earlier inability to isolate a opiate receptor messenger RNA by reverse transcriptase-polymerase chain reaction using a human primer. This suggests that a novel opiate receptor was possibly present and selective toward M6G.
The parasitic worm Ascaris suum contains the opiate alkaloid morphine as determined by HPLC coupl... more The parasitic worm Ascaris suum contains the opiate alkaloid morphine as determined by HPLC coupled to electrochemical detection and by gas chromatography/mass spectrometry. The level of this material is 1168 ؎ 278 ng/g worm wet weight. Furthermore, Ascaris maintained for 5 days contained a significant amount of morphine, as did their medium, demonstrating their ability to synthesize the opiate alkaloid. To determine whether the morphine was active, we exposed human monocytes to the material, and they immediately released nitric oxide in a naloxone-reversible manner. The anatomic distribution of morphine immunoreactivity reveals that the material is in the subcuticle layers and in the animals' nerve chords. Furthermore, as determined by RT-PCR, Ascaris does not express the transcript of the neuronal receptor. Failure to demonstrate the expression of this opioid receptor, as well as the morphine-like tissue localization in Ascaris, suggests that the endogenous morphine is intended for secretion into the microenvironment.
Both morphine and anandamide significantly stimulated cultured endothelial intracellular calcium ... more Both morphine and anandamide significantly stimulated cultured endothelial intracellular calcium level increases in a concentration-dependent manner in cells pre-loaded with fura 2rAM. Morphine Ž w x. w x is more potent than anandamide approximately 275 vs. 135 nM Ca , and the Ca for both ligands was i i blocked by prior exposure of the cells to their respective receptor antagonist, i.e., naloxone and SR 171416A. Various opioid peptides did not exhibit this ability, indicating a morphine-mediated process. In comparing 3 w x the sequence of events concerning morphine's and anandamide's action in stimulating both Ca and nitric i oxide production in endothelial cells, we found that the first event precedes the second by 40 " 8 sec. The w x opiate and cannabinoid stimulation of Ca was attenuated in cells leeched of calcium, strongly suggesting that i intracellular calcium levels regulate cNOS activity. CELL SIGNAL 11;3:189᎐193, 1999.
Morphine and anandamidc stimulate the release of nitric oxide ~,NO) in diverse tissues. Tile pres... more Morphine and anandamidc stimulate the release of nitric oxide ~,NO) in diverse tissues. Tile present study examines file consequences of this action on neumtransmittcr release in ganglia from two inverlebrates: ventral eL:m,' ganglia ftum tile leech flirudo medicinali.s and the pedal ganglion fi'om the mussel Mvtilus e&dis, in Ihese ganglia, preloaded serot,',mn (5-HT) and dopamme (DA) can be released by 50 mM KCI. Anandamide, an endogenous cannabinoid substance, suppresses tile pota,sium-stimulated release or' [ ~H]DA (80~/,), but not 5-HT, in a ctmcentratmn-dependent manner, lrom the neural tissues of both. The cflZ-ct of anandamide can be antagonized by pre-exposing the neural tissues of both animals to SR 141716A, a potent cannabinoid receptor antagonist. Prior treatment of the ganglia with N-~-nitm-t.-argininc methyl ester (|,+NAME). a nitric oxide synthase inhibito;, significantly diminishes the inhibitory effect of anandanfide. Morphine also inhibits [~|-|ll)A l'eleas~2 H| a |ialoxollc.. and t-NAME+sensitive manner. Anandamide and morphhlc act through separate mechanisms since the respective antagonists show no cross r+. tctivily. The N() dollor, SNAP, depressed the potassiunl-sthnuhtled rele-~se ',,f plreh~at|ed [ +till)A, but !1ol 5-HT, in the neural n;,, s of both animals. R)-Ala,-Mcts enkephalinamidc (II)AMA) also inhibited the po,as.,,ium-stinmlated reteasc ol [ ~|-|]|)A in a nahmmc+scnsitiv¢ process. However, file el feet of DAMA was seen in tile presence of I,+NAME (1(} a M), indicatin~ thai the ophud pei'btidc inhihi|ion t'Jl |he i','csymq'~tic relea~,e of I)A in not coupled IO NO. We postulate that cannabinolds and tlleir endogenous eft ectors play a prominent role in the regulation t)t catecholamine tcJea,',c ill invertebnttes via NO release as is the case for opiate alkaloids. ~' 1997 Elsevier Science B.V.
The present study demonstrates that morphine-and codeine-like molecules are present in Schistosom... more The present study demonstrates that morphine-and codeine-like molecules are present in Schistosoma mansoni following HPLC separation and identification with. an appropriate commercially available antibody. Furthermore, the endogenous material, corresponding to ...
The tissue distribution, course of secretion, and sex differences of morphine were delineated in ... more The tissue distribution, course of secretion, and sex differences of morphine were delineated in Ascaris suum. Nitric oxide (NO) release in various tissues in response to morphine and its metabolite morphine-6-glucuronide (M6G) were also examined. Ascaris suum of both sexes along with their incubation fluid were analyzed for morphine concentrations by high- performance liquid chromatography (HPLC) over a 5-day period. Various tissues were also dissected for HPLC and NO analysis. Morphine was found to be most prevalent in the muscle tissue, and there is significantly more morphine in females than males, probably because of the large amounts present in the female uterus. Morphine (10−9 M) and M6G (10−9 M) stimulated the release of NO from muscles. Naloxone (10−7 M) and N-nitro-l-arginine methyl ester (10−6 M) blocked (P < 0.005) morphine-stimulated NO release from A. suum muscle tissue. d-Phe-Cys-Tyr-d-Trp-Om-Thr-Pen-Thr-NH2 (CTOP) (10−7 M) did not block morphine's NO release. ...
The effects of the opiates morphine and morphine-6-glucuronide (M6G), the μ opioid receptor speci... more The effects of the opiates morphine and morphine-6-glucuronide (M6G), the μ opioid receptor specific antagonist D-Phe-Cys-Tyr-D-Trp-Om-Thr-Pen-Thr-NH 2 (CTOP), and the general opiate antagonist naloxone on the latency of response to thermal stimulation were determined in the parasitic nematode Ascaris suum. Thermal detection and avoidance behaviors of the worms were evaluated with a tail flick analgesia meter using a modification of a technique employed for nociception experiments in rodents. Morphine and M6G were shown to have a dose dependent analgesic effect on A. suum's latency of response to heat with morphine being the most potent. The analgesic effect of morphine was reversed by naloxone but not CTOP. Neither naloxone nor CTOP was able to block the analgesia of M6G. CTOP but not naloxone had significant analgesic effects on its own. These findings are generally consistent with previous results on the effects of opiates and nitric oxide release from A. suum tissue. Apparently these nematodes possess opioid receptors that effect nociception.
Samples of mosquitoes in the Culex pipiens L. complex from Memphis, Tenn., were collected from Ju... more Samples of mosquitoes in the Culex pipiens L. complex from Memphis, Tenn., were collected from June to November 1985 and examined in regard to allozyme frequencies and ratios of the two arms of the phallosome of the male genitalia (DV/D). The dominant allozymes of hexokinase (HkA) and 6-phosphogluconate dehydrogenase (PgdF) significantly increased in frequency during this period as did the mean DV/D ratio. An analysis of gene frequencies by species group designated by DV/D ratios revealed no significant differences among Cx. pipiens, Cx. quinquefasciatus Say, and intermediates. The lack of association of gene frequencies with the taxa determined by the DV/D ratio indicated that although allozyme frequencies were correlated temporally with the DV/D ratio in the population, they were not associated with subspecies. These results are consistent with previous work that has shown latitudinal association and thermal stability differences in the major allozymes of these enzymes in the Cx. ...
Medical science monitor : international medical journal of experimental and clinical research, 2005
The current paper summarizes the findings on endogenous morphine isolated by HPLC and characteriz... more The current paper summarizes the findings on endogenous morphine isolated by HPLC and characterized by mass spectroscopy in Schistosoma mansoni, Dracunculus medinensis and Ascaris suum. Morphine-6-glucuronide (M6G) has also been found by HPLC and confirmed by mass spectroscopy in Dracunculus medinensis and Ascaris suum. In addition, a morphine like substance has been isolated from Trichinella spiralis by HPLC and mice infected with Trichinella spiralis show a naloxone reversible analgesia. We discuss in greater detail the tissue distribution, course of secretion, and sex differences of morphine in Ascaris suum. Finally, we explore the function of morphine as both an internal signaling molecule and its use in immune evasion in Ascaris suum.
The effects of the opiates morphine and morphine-6-glucuronide (M6G), the mu opioid receptor spec... more The effects of the opiates morphine and morphine-6-glucuronide (M6G), the mu opioid receptor specific antagonist D-Phe-Cys-Tyr-D-Trp-Om-Thr-Pen-Thr-NH(2) (CTOP), and the general opiate antagonist naloxone on the latency of response to thermal stimulation were determined in the parasitic nematode Ascaris suum. Thermal detection and avoidance behaviors of the worms were evaluated with a tail flick analgesia meter using a modification of a technique employed for nociception experiments in rodents. Morphine and M6G were shown to have a dose dependent analgesic effect on A. suum's latency of response to heat with morphine being the most potent. The analgesic effect of morphine was reversed by naloxone but not CTOP. Neither naloxone nor CTOP was able to block the analgesia of M6G. CTOP but not naloxone had significant analgesic effects on its own. These findings are generally consistent with previous results on the effects of opiates and nitric oxide release from A. suum tissue. Appar...
R ECENT studies have used the blue mussel, Mytilus edulis, as a sentinel organism for pollution (... more R ECENT studies have used the blue mussel, Mytilus edulis, as a sentinel organism for pollution (Greig & Sennefelder 1985; Micallef & Tyler 1989). The mussel and other marine mollusks, such as oysters and clams, have long been known to concentrate toxic heavy metals, such as mercury, lead and cadmium. They are also capable of bioaccumulating synthetic organic compounds, such as PCBs and pesticides. This propensity to concentrate toxins often poses a public health problem, but it also offers an opportunity for environmental monitoring. The United Nations Environmental Program has in fact launched a global effort to monitor these organisms for marine pollution (Bayne 1989; Beeby 1993; Borchart et al. 1988). Not only can mussels act as reservoirs to measure the extent of chemical contamination and its variation in different coastal environments, it can also be used in the teaching lab to illustrate biological responses to pollution (Bayne 1989; Conrad 1988). Pollutants, such as heavy metals, have been found to alter molecular and cellular processes in mussels. While each organism is unique in both its capacity to deal with pollutants and its biological responses to them, certain cellular responses are often similar down the phylogenetic scale. A serious concern in recent years is the immune suppressive effect that pollutants are having on marine life. This phenomenon has been observed from invertebrate organisms, such as mussels and oysters, to vertebrates such as fish, seals and whales (De L. Swart et al. 1994; Faisal et al. 1991; Sami et al. 1992). The blue mussel offers an easy way of introducing your class to both immune response and the effects of environmental pollution on marine organisms. The immune cells of mollusks are called hemocytes and can be easily extracted with hemolymph from the mussel's foot or heart with a syringe. These cells have been shown to have many characteristics of vertebrate granular leukocytes. They respond to the same mes-
The tissue distribution, course of secretion, and sex differences of morphine were delineated in ... more The tissue distribution, course of secretion, and sex differences of morphine were delineated in Ascaris suum. Nitric oxide (NO) release in various tissues in response to morphine and its metabolite morphine-6-glucuronide (M6G) were also examined. Ascaris suum of both sexes along with their incubation fluid were analyzed for morphine concentrations by highperformance liquid chromatography (HPLC) over a 5-day period. Various tissues were also dissected for HPLC and NO analysis. Morphine was found to be most prevalent in the muscle tissue, and there is significantly more morphine in females than males, probably because of the large amounts present in the female uterus. Morphine (10 Ϫ9 M) and M6G (10 Ϫ9 M) stimulated the release of NO from muscles. Naloxone (10 Ϫ7 M) and N-nitro-L-arginine methyl ester (10 Ϫ6 M) blocked (P Ͻ 0.005) morphine-stimulated NO release from A. suum muscle tissue. D-Phe-Cys-Tyr-D-Trp-Om-Thr-Pen-Thr-NH 2 (CTOP) (10 Ϫ7 M) did not block morphine's NO release. However, naloxone could not block M6G-stimulated NO release by muscles, whereas CTOP (10 Ϫ7 M) blocked its release. These findings were in seeming contradiction to our earlier inability to isolate a opiate receptor messenger RNA by reverse transcriptase-polymerase chain reaction using a human primer. This suggests that a novel opiate receptor was possibly present and selective toward M6G.
The parasitic worm Ascaris suum contains the opiate alkaloid morphine as determined by HPLC coupl... more The parasitic worm Ascaris suum contains the opiate alkaloid morphine as determined by HPLC coupled to electrochemical detection and by gas chromatography/mass spectrometry. The level of this material is 1168 ؎ 278 ng/g worm wet weight. Furthermore, Ascaris maintained for 5 days contained a significant amount of morphine, as did their medium, demonstrating their ability to synthesize the opiate alkaloid. To determine whether the morphine was active, we exposed human monocytes to the material, and they immediately released nitric oxide in a naloxone-reversible manner. The anatomic distribution of morphine immunoreactivity reveals that the material is in the subcuticle layers and in the animals' nerve chords. Furthermore, as determined by RT-PCR, Ascaris does not express the transcript of the neuronal receptor. Failure to demonstrate the expression of this opioid receptor, as well as the morphine-like tissue localization in Ascaris, suggests that the endogenous morphine is intended for secretion into the microenvironment.
Both morphine and anandamide significantly stimulated cultured endothelial intracellular calcium ... more Both morphine and anandamide significantly stimulated cultured endothelial intracellular calcium level increases in a concentration-dependent manner in cells pre-loaded with fura 2rAM. Morphine Ž w x. w x is more potent than anandamide approximately 275 vs. 135 nM Ca , and the Ca for both ligands was i i blocked by prior exposure of the cells to their respective receptor antagonist, i.e., naloxone and SR 171416A. Various opioid peptides did not exhibit this ability, indicating a morphine-mediated process. In comparing 3 w x the sequence of events concerning morphine's and anandamide's action in stimulating both Ca and nitric i oxide production in endothelial cells, we found that the first event precedes the second by 40 " 8 sec. The w x opiate and cannabinoid stimulation of Ca was attenuated in cells leeched of calcium, strongly suggesting that i intracellular calcium levels regulate cNOS activity. CELL SIGNAL 11;3:189᎐193, 1999.
Morphine and anandamidc stimulate the release of nitric oxide ~,NO) in diverse tissues. Tile pres... more Morphine and anandamidc stimulate the release of nitric oxide ~,NO) in diverse tissues. Tile present study examines file consequences of this action on neumtransmittcr release in ganglia from two inverlebrates: ventral eL:m,' ganglia ftum tile leech flirudo medicinali.s and the pedal ganglion fi'om the mussel Mvtilus e&dis, in Ihese ganglia, preloaded serot,',mn (5-HT) and dopamme (DA) can be released by 50 mM KCI. Anandamide, an endogenous cannabinoid substance, suppresses tile pota,sium-stimulated release or' [ ~H]DA (80~/,), but not 5-HT, in a ctmcentratmn-dependent manner, lrom the neural tissues of both. The cflZ-ct of anandamide can be antagonized by pre-exposing the neural tissues of both animals to SR 141716A, a potent cannabinoid receptor antagonist. Prior treatment of the ganglia with N-~-nitm-t.-argininc methyl ester (|,+NAME). a nitric oxide synthase inhibito;, significantly diminishes the inhibitory effect of anandanfide. Morphine also inhibits [~|-|ll)A l'eleas~2 H| a |ialoxollc.. and t-NAME+sensitive manner. Anandamide and morphhlc act through separate mechanisms since the respective antagonists show no cross r+. tctivily. The N() dollor, SNAP, depressed the potassiunl-sthnuhtled rele-~se ',,f plreh~at|ed [ +till)A, but !1ol 5-HT, in the neural n;,, s of both animals. R)-Ala,-Mcts enkephalinamidc (II)AMA) also inhibited the po,as.,,ium-stinmlated reteasc ol [ ~|-|]|)A in a nahmmc+scnsitiv¢ process. However, file el feet of DAMA was seen in tile presence of I,+NAME (1(} a M), indicatin~ thai the ophud pei'btidc inhihi|ion t'Jl |he i','csymq'~tic relea~,e of I)A in not coupled IO NO. We postulate that cannabinolds and tlleir endogenous eft ectors play a prominent role in the regulation t)t catecholamine tcJea,',c ill invertebnttes via NO release as is the case for opiate alkaloids. ~' 1997 Elsevier Science B.V.
The present study demonstrates that morphine-and codeine-like molecules are present in Schistosom... more The present study demonstrates that morphine-and codeine-like molecules are present in Schistosoma mansoni following HPLC separation and identification with. an appropriate commercially available antibody. Furthermore, the endogenous material, corresponding to ...
Uploads
Papers by Stephen Pryor