Effect of Pethidine Hydrochloride on the Development of Neural Tube: A Genetic Analysis Study in a Chick Embryo Model.

BACKGROUND: Neural tube defects are among the most frequent congenital abnormalities of the central nervous system. Progression of neural tube deficits is affected by hereditary predilection and environmental determinants. Pethidine (meperidine) is a fast and powerful opioid analgesic in U.S. Food and Drug Administration category C. There are reports about developmental anomalies due to this medication. The aim of this study was to investigate the effects of different doses of pethidine hydrochloride on neural tube development in a chick embryo model resembling the first month of vertebral growth in mammals. METHODS: Seventy-five specific pathogen-free eggs were incubated for 28 hours and divided into 5 groups (including the control group), each consisting of 15 eggs. Pethidine hydrochloride was administered sub-blastodermically with a Hamilton microinjector in 4 different doses. Incubation was continued until the end of the 48th hour. Subsequently, all eggs were opened, and embryos were cut from the embryonic membranes and evaluated morphologically, genetically, and histopathologically. RESULTS: Crown-rump length, somite numbers, and silver-stained nucleolar organizer region (AgNOR) number averages, and total AgNOR/nuclear area ratios decreased in a dose-dependent manner. Examination of neural tube closure revealed statistically significant differences in all experimental groups (P<0.05). Messenger RNA levels of the BRE gene were decreased in pethidine hydrochloride-exposed embryos compared with the control group. Although this downregulation was not statistically significant, this decrease was striking with a 0.422-fold change in the fifth group. CONCLUSIONS: We demonstrated that pethidine hydrochloride affects neuronal development in chicken embryos. The teratogenic mechanism of pethidine hydrochloride is unclear; therefore, further investigation is required.

Suspected acute meperidine toxicity in a dog.

OBSERVATIONS: A 22-month-old male neutered Coton De Tulear dog was presented for upper gastrointestinal endoscopy under general anesthesia. The anesthetic plan included premedication with intramuscular meperidine (4 mg kg(-1)) but meperidine was inadvertently administered at ten-fold this dose. Within 5 minutes, the dog was unresponsive to external stimulation, and by 10 minutes post-injection developed generalized signs of central nervous system (CNS) excitement. Initial therapy included inspired oxygen supplementation, and single intravenous (IV) doses of diazepam (0.68 mg kg(-1)) and naloxone (0.03 mg kg(-1)) to no effect. A second dose of diazepam (0.46 mg kg(-1), IV) abolished most of the signs of CNS excitement. General anesthesia was induced and the endoscopy performed. Time to extubation was initially prolonged, but administering naloxone (final dose 0.1 mg kg(-1), IV) to effect enabled extubation. After naloxone, the dog became agitated, noise sensitive, and had leg and trunk muscle twitches. Diazepam (0.30 mg kg(-1), IV) abolished these signs and the dog became heavily sedated and laterally recumbent. Naloxone administration was continued as a constant rate infusion (0.02 mg kg(-1) hour(-1), IV) until approximately 280 minutes post-meperidine injection, at which time the dog suddenly sat up. Occasional twitches of the leg and trunk muscles were observed during the night. The dog was discharged the next day appearing clinically normal. CONCLUSIONS: Given that the CNS excitatory effects of normeperidine are not a mu opioid receptor effect, the use of naloxone should be considered carefully when normeperidine excitotoxicity is suspected. Benzodiazepines may be beneficial in ameliorating clinical signs of normeperidine excitotoxicity.

[Lethal intoxication while driving a car]. / Letale Intoxikation im fahrenden PKW.

This report concerns a passenger car crash, which at first looked like a case of multiple trauma resulting in the death of the pinned-in driver. Conspicuous was a cuff that had been applied to the left forearm. No significant injuries could be determined during the autopsy of the corpse. The forensic toxicology results showed lethal toxic concentrations of the painkiller pethidine, which suggests that the driver had applied a lethal dose of the medication immediately before the collision as part of a complex suicide.

The interaction between monoamine oxidase inhibitors and narcotic analgesics in mice.

1. The administration of either iproniazid or tranylcypromine to mice potentiates the acute toxicity of pethidine, morphine, pentazocine and phenazocine.2. Blood levels of pentazocine in mice pretreated with tranylcypromine do not differ from the levels in animals not receiving the monoamine oxidase (MAO) inhibitor.3. There is no correlation between changes in brain and liver MAO activity and the increased pethidine toxicity.4. A comparison is made between the change in pethidine toxicity and the changes in the concentration of cerebral noradrenaline, dopamine and 5-hydroxytryptamine following the injection of tranylcypromine.5. It is concluded that the increased toxicity of potent analgesics in combination with MAO inhibitors is not due to a decelerated metabolism of the analgesic drug, but is related to an increased concentration of cerebral 5-hydroxytryptamine. A critical level of this monoamine, in the brain, may be necessary before the drug interaction will take place.

Effect of nonselective and selective inhibitors of monoamine oxidases A and B on pethidine toxicity in mice.

The LD50 of pethidine was determined in mice pretreated (4 h) either with the nonselective monoamine oxidase (MAO) inhibitor, phenelzine or with clorgyline, a selective inhibitor of MAO A or deprenyl, a selective inhibitor of MAO B. Phenelzine or combined clorgyline plus deprenyl pretreatments decreased pethidine LD50. Clorgyline or deprenyl alone did not affect pethidine toxicity. Whole brain 5-hydroxytryptamine (5-HT) concentrations were measured in the pretreated mice. 5-HT levels were approximately doubled (P less than 0.001) after phenelzine or clorgyline plus deprenyl treatment, but not after clorgyline or deprenyl given alone. These results indicate that both MAO A and MAO B need to be inhibited to increase pethidine toxicity and brain 5-HT levels. They support the involvement of 5-HT in the toxic interaction between pethidine and MAO inhibitors.

Comparison of the effects of morphine, pethidine and pentazocine in rabbits pretreated with a monoamine oxidase inhibitor.

1. Rabbits were premedicated with pargyline and the changes in rectal temperature measured after the intravenous infusion of morphine, pentazocine and pethidine.2. Pethidine produced pronounced rises in rectal temperature which were dose dependent. One out of the four rabbits given 1 mg/kg died in hyperthermia. Four out of the four rabbits given 5 mg/kg died in hyperthermia. Doses of 10 mg/kg of pethidine caused no significant change in the rectal temperature of rabbits not pretreated with pargyline.3. Morphine and pentazocine in doses of 1 mg and 10 mg/kg did not significantly alter the rectal temperature of rabbits pretreated with pargyline except for one rabbit which developed a delayed hyperthermia following the injection of morphine 1 mg/kg.

Relative cataleptic potency of narcotic analgesics, including 3,6-dibutanoylmorphine and 6-monoacetylmorphine.

Dose-response curves were constructed for duration of significant catalepsy in male Sprague-Dawley rats following i.p. administration of four established and two novel narcotic analgesics. The relative potency of the six drugs was levorphanol greater than monacetylmorphine greater than diacetylmorphine greater than dibutanoylmorphine greater than morphine greater than meperidine. Cataleptic potency correlated with analgesic potency for morphine, diacetylmorphine, and dibutanoylmorphine but not for monoacetylmorphine.

Behavioural effects of norpethidine, a metabolite of pethidine, in rats.

This study investigated behavioural effects of the toxic pethidine metabolite, norpethidine, in rats and its interactions with reserpine, apomorphine and physostigmine. Following intraperitoneal administration, brain concentrations of norpethidine reached a plateau after 20-40 min and remained elevated for 2 h. In the dose range 0.06-0.18 mmol/kg, norpethidine induced myoclonic jerks, a characteristic splayed posture, and episodes of exaggerated shivering. Forward locomotion, grooming, yawning and rearing were suppressed. Seizures and reverse locomotion occurred occasionally. Administration of reserpine 1 h prior to norpethidine, or of apomorphine or physostigmine 15 min after norpethidine, did not alter the norpethidine-induced behaviours; neither did norpethidine block the effects of apomorphine or physostigmine. The characteristic profile of behaviours induced by norpethidine make this toxicant readily amenable to animal studies. Our results indicate that its mechanism of action is unlikely to involve dopaminergic or cholinergic pathways.

Modification by monoamine oxidase inhibitors of the analgesic, hypothermic and toxic actions of morphine and pethidine in mice.

A single injection of phenelzine 100 mg kg-1 given 18 h before, decreased the analgesia and hypothermia induced by morphine, but potentiated the analgesic and hypothermic effects of pethidine, when the analgesics were administered either intraperitoneally, or intracerebroventricularly. The modification of pethidine analgesia and hypothermia, but not morphine analgesia, was antagonized by methysergide (10 mg lg-1, s.c.). The LD50 of pethidine, but not that of morphine, was 30-40% lower in mice treated with phenelzine tranylcypromine or iproniazid 6 h before the test. The increased lethality of a single dose of pethidine induced by phenelzine was also prevented by methysergide. Pretreatment of mice with 100 mg kg-1 phenelzine was followed by a significant rise in both brain tryptophan and 5-hydroxytryptamine (5-HT) concentrations which lasted for 24 h. Therefore, the changes in pethidine effects could have been due to raised brain tryptophan and 5-HT concentrations.

The effect of drugs on the mortality of mice inoculated with Friend leukaemia virus or toxoplasma gondii.

Infection and tumors provoke substantial changes accompanied with the disbalance of many neuroendocrine factors which in their summarizing effects influence the life span of animals. Our previous results showed enhanced mortality after one injection of morphine in association with Friend leukaemia virus infection. The aim of this study was to examine the effects of some other opioids (pethidine and pentazocine) and an acetylcholine esterase inhibitor neostigmine on the survival of animals under two conditions: (1) Friend leukaemia virus infection which mostly depressed immune functions, and (2) Toxoplasma gondii infection which in general enhanced the immune status. In contrast to our previous observation with morphine, the mortality induced by single doses of pethidine (150 mg/kg) or pentazocine (50-75 mg/kg) was unchanged during the Friend leukaemia virus infection. A single injection of neostigmine (0.42 or 0.56 mg/kg) was significantly more lethal in DBA-2 mice infected with Friend leukaemia virus. Neostigmine in doses of 0.33 and 0.4 mg/kg caused death in 46 % and 57 %, respectively, of animals infected with Toxoplasma gondii which was significantly higher in comparison with only 8 % and 12.5 % in control groups. Pethidine (150 mg/kg) killed 70 % of Toxoplasma gondii infected animals and even 90 % of non-infected mice. Thus, the Friend leukaemia virus and Toxoplasma gondii infections increased toxicity only of some drugs which may, at least partly, be associated with altered immune status during infection and involvement of the cholinergic system.

Interaction of butorphanol, with monoamine oxidase inhibitor, tranylcypromine.

This investigation examines the possibility of interaction between tranylcypromine and butorphanol in comparison to pethidine. The LD50 of pethidine and butorphanol were determined in mice pretreated with the non-selective monoamine oxidase (MAO) inhibitor, tranylcypromine orally for 8 days or with oral saline solution. Tranylcypromine decreased the LD50 of both pethidine and butorphanol by 78% and 41%, respectively. Anesthetized rabbits with halothane pretreated with tranylcypromine or saline were given pethidine (5 mg/kg i.v.) or butorphanol (0.5, 1 and 2 mg/kg i.v.). Pethidine produced a marked increase in blood pressure in rabbits pretreated with tranylcypromine and did not affect significantly the heart rate. Butorphanol did not affect either blood pressure or heart rate at doses of 0.5 or 1 mg/kg. However, the largest dose of butorphanol (2 mg/kg) produced hypotension and tachycardia in rabbits pretreated with tranylcypromine. Neither pethidine nor butorphanol affected the temperature of anesthetized rabbits pretreated with tranylcypromine or saline.

Meperidine and normeperidine metabolism in the rhesus monkey.

Meperidine and its principle metabolite, normeperidine, were given intravenously to four non-human primates prior to cesarean delivery in an equivalent dose for human parturients. The status of the infants regarding neonatal depression was assessed at delivery. Repeated blood samples from both the mother and the neonate were obtained over a period of 4 days. The levels of meperidine and normeperidine were analyzed. The results showed that the metabolism of meperidine and normeperidine in the non-human primate was essentially the same as that observed in the human parturient. In addition, normeperidine appeared to be more toxic than meperidine to the neonate. Finally, there does not appear to be an evidence for neonatal metabolism of meperidine to normeperidine.

Interaction between ethanol and opioids in a protozoan assay.

Ethanol excess combined with opioids can be fatal due to their toxic interaction, yet the nature of the interaction is little documented. Since ethanol and some opioids have membrane stabilizing activity, the present study used a protozoan motility model to test the possibility that ethanol may interact with some opioids on this basis. The EC50 in motility reduction for ethanol, dextropropoxyphene, methadone and pethidine was 522.0 +/- 36.7 mM, 0.59 +/- 0.08 mM, 0.40 +/- 0.09 mM and 4.57 +/- 0.36 mM, respectively. When ethanol was combined with one of the three drugs in equitoxic doses at a ratio of 0.5:0.5, the predicted/observed EC50 values for ethanol-dextropropoxyphene, ethanol-methadone and ethanol-pethidine were 1.37, 1.11 and 1.00, each being close to unity, indicating an additive interaction. The interaction between ethanol and dextropropoxyphene was further explored at 0.25:0.75 and 0.75:0.25 equitoxic dose ratios, with the predicted/observed EC50 values of 0.98 and 0.97, also showing an additive interaction. This suggests that a non-specific interaction between ethanol and opioids may also take place in vivo, which could cause increased toxicity over and above the involvement of opioid receptors. Information from this study should aid understanding of the mechanism of interactions in human poisoning by agents with membrane stabilizing activity.

The effect of chronic toxicity of pethidine on the spinal cord: an experimental model in rabbits.

The aim of this study was to evaluate the toxicity of chronic spinal analgesia with pethidine in a rabbit model. We introduced epidural catheters in twenty New Zealand white rabbits, divided into two groups, and we administered 0.5 mg/kg pethidine or the same volume of normal saline through the catheters, for three consecutive days. Throughout the experiment, the animals were evaluated in terms of neurological status using the Tarlov score. After the rabbit's euthanasia, 4 µm sections of spinal cord stained with Hematoxylin-Eosin were analyzed by a pathologist blinded to the study for neurohistopathological changes. The results were statistically analyzed with Prism 5 software for Windows. No significant differences were noticed between the two groups in as far as body temperature (p=0.295) and weight (p=0.139) were concerned. In the group of animals, which received epidural pethidine, nine rabbits showed histological changes suggestive for neurotoxicity at the lumbar level of the spinal cord. These findings were significantly different compared with the control group which received only saline (no microscopic lesions revealed; p=0.0006). When combining the data from both groups or using the pethidine group alone, there was a significant correlation between the presence of neurological injury (Tarlov score) and the presence of the histopathological lesions in the spinal cord (r=-0.709, p=0.0002 and r=-0.635, p=0.013, respectively). Based on our findings, the chronic epidural administration of pethidine in rabbits induces moderate to severe histological changes on the spinal cord, but further investigations are needed to make a definitive statement about the histological effect of pethidine on the neurological tissue.

Examination of the effect of dose-death interval on detection of meperidine exposure in decomposed skeletal tissues using microwave-assisted extraction.

The effect of dose-death interval and tissue distribution on the detection of meperidine in selected skeletal tissues was examined using a rapid microwave-assisted extraction (MAE) methodology. Rats (n=14) were dosed with 0 (n=2) or 30 mg/kg (n=12) meperidine (i.p.). Drug-positive rats were sacrificed with CO(2) after 20, 30, 90 and 150 min (n=3 per group). Heart blood was collected immediately after death. Tibiae were excised and frozen for further analysis. The remaining carcasses were allowed to decompose outside in secured cages to the point of complete skeletonization in a rural Northern Ontario location during the late summer months. Vertebrae and pelvi were collected for each animal. Tibial marrow was homogenized in 3 mL PB6 (phosphate buffer, 0.1M, pH 6). Fresh tibiae, and decomposed vertebrae and pelvi were cleaned in PB8.5 (phosphate buffer, 0.1M, pH 8.5) and sonicated to remove remaining soft tissue. Samples of dried, ground bone (0.5-1g) suspended in 2 mL PB6 were then irradiated in a domestic microwave oven (1100 W) at atmospheric pressure for 15 min. Samples of vertebral bone (1g) were also extracted by passive incubation in methanol (3 mL, 50°C, 72 h). All supernatants then underwent solid-phase extraction and analysis by GC/MS, using electron impact ionization in the Selected Ion Monitoring (SIM) mode. Mean GC/MS responses for each tissue type were negatively correlated with dose-death interval, with correlation coefficients ranging from -0.32 to -0.87. Analysis of variance showed dose-death interval to be a main effect (p<0.05) with respect to GC/MS response for blood, marrow, tibial epiphyses prepared by MAE, and vertebral bone prepared by passive extraction, but not for tibial diaphyses, pelvi or vertebrae prepared by MAE. Overall, MAE is advantageous as a rapid extraction tool for screening purposes in skeletal tissues, but assignment of significance to quantitative expressions of skeletal drug concentrations is complex and should be approached with caution.