Medical linear accelerator mounted mini-beam collimator: design, fabrication and dosimetric characterization.

The goal of this work was to design, build and experimentally characterize a linear accelerator mounted mini-beam collimator for use at a nominal 6 MV beam energy. Monte Carlo simulation was used in the design and dosimetric characterization of a compact mini-beam collimator assembly mounted to a medical linear accelerator. After fabrication, experimental mini-beam dose profiles and central axis relative output were measured and the results used to validate the simulation data. The simulation data was then used to establish traceability back to an established dosimetric code of practice. The Monte Carlo simulation work revealed that changes in collimator blade width have a greater influence on the valley-to-peak dose ratio than do changes in blade height. There was good agreement between the modeled and measured profile data, with the exception of small differences on either side of the central peak dose. These differences were found to be systematic across all depths and result from limitations associated with the collimator fabrication. Experimental mini-beam relative output and simulation data agreed to better than ± 2.0%, which is well within the level of uncertainty required for dosimetric traceability of non-standard field geometries. A mini-beam collimator has now been designed, built and experimentally characterized for use with a commercial linear accelerator operated at a nominal 6 MV beam energy.

The effects of nandrolone decanoate on cocaine-induced kindling in male rats.

Use and abuse of various controlled substances in recent years has reached alarming levels. Among these are cocaine and anabolic steroids. The two contrasting types of drug have common sites of action within the limbic system of the central nervous system. The ability of cocaine to provoke seizures is well documented, and sex hormones also have been shown to alter seizure types and characteristics. This project studied the consequences of co-administration of cocaine and a representative anabolic-androgenic steroid, nandrolone decanoate. Specifically, the effects of nandrolone on cocaine-induced kindling of seizures were examined. Nandrolone was shown to increase seizure rate when given in high (20mg twice weekly) intermittent doses. No statistically significant differences were observed with low (2mg) daily doses of nandrolone. The results support the hypothesis that an androgen may interact so as to modify the pattern of cocaine-related kindling. However, the potential of either pharmacodynamic and/or pharmacokinetic mechanism(s) for this interaction exists, and the nature of these interactions remains to be fully elucidated.

Time-dose relationships for locomotor activity effects of morphine after acute or repeated treatment.

1. Effects of morphine sulphate (1.25, 2.5, 5, 10, 20 and 40 mg/kg i.p.) on locomotor activity of male rats were observed for 8 h after single doses in non-tolerant rats. The lower three doses had only an excitatory effect, whereas the higher three doses caused initial depression followed by a delayed excitatory effect.2. The same doses of morphine were administered daily for 30 days. No tolerance developed within this time to the excitatory effect. The locomotor excitatory effect of the higher three doses of morphine became progressively more pronounced over treatment periods of 30 days (and 48 days for 20 mg/kg), while the latency to peak activity decreased.3. An explanation of these results is suggested on the basis of two different central drug-receptor interactions affecting motility.

Freezing action of a metabolite of haloperidol in frogs.

In vivo metabolism studies led to the identification of a previously proposed metabolite of haloperidol, 4-(4'-chlorophenyl)-4-piperidinol (CPPO), in the liver of a haloperidol-treated rat. However, the secondary metabolites of CPPO that we have proposed were not observed in this study. Neurotoxicity studies in frogs, which have been used to detect N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) action, showed that CPPO did not mimic the neurotoxicity of MPTP but caused a delayed and persistent freezing action in Rana pipiens frogs. It is proposed that this action may contribute to some of the delayed side-effects associated with haloperidol therapy.

Alcohol-associated conditioned reinforcement.

Research was conducted to examine the ability of alcohol to impart conditioned reinforcement. Rats were allowed to self-administer solutions of either saline or alcohol (25, 50, and 100 mg/kg/infusion) by the intragastric route. Superimposed on the infusion interval was a buzzer (conditioned reinforcing stimulus). Tests during extinction revealed that conditioned reinforcement had been acquired. Results also indicated that as the paired unit dose was increased, potency of the conditioned reinforcer increased. In a second study, the lever-pressing response, which produced saline infusion and the buzzer, became available only subsequent to 5 sessions of pairing the buzzer with infusions of saline or alcohol. The results indicated that lever pressing increased with increasing unit dosage of alcohol infusions in prior pairings.

Effect of unit dose and route of administration on self-administration of morphine.

Rats were implanted with intravenous or intragastric cannulas and allowed to self-administer morphine sulfate in doses of 0 (saline), 0.03, 0.1, 0.3, 1.0, 3.0, and 10.0 mg/kg/infusion. For the intravenous route the number of infusions decreased with increasing unit dose, while the amount self-administered was directly related to unit dose. However, for the intragastric route the number of infusions first increased and then decreased as unit dose was elevated, while the amount self-administered again increased with unit dose. Comparisons between routes showed that for intragastric subjects the number of infusions and amount self-administered both were lower at the two lowest doses but higher for all other doses. These results support the expectation that intravenous injection should produce more potent reinforcing effects than intragastric administration.

Morphologic aspects of experimental esophageal lye strictures. II. Effect of steroid hormones, bougienage, and induced lathyrism on acute lye burns.

Among 77 dogs surviving standardized transmural esophageal lye injury for at least 2 weeks and as long as 12 weeks, 24 were untreated, 26 received corticosteroids and bougienage (S&B), and 27 received only the lathyrogen beta-aminoproprionitrile (BAPN). Stricture frequency was reduced markedly and significantly in the S&B and BAPN groups when compared to the controls (p less than 0,01). Strictures resulted from inward circumferential remodeling of all mural layers, not proliferating bulky scar tissue, and persistent ulceration was apparently not an influential factor in any group. The S&B dogs invariably showed reduction of the internal or mucosal length of the injured segment as compared to the outer length; these relations were quite variable in the other two groups so that mean internal shortening was significantly greater (p less than 0.01) in the S&B group. Marked mural thinning in the injured zone was present in all three groups but was most frequent in the BAPN-treated animals. The major conclusion is that BAPN-induced changes in the physical properties of reparative tissue can increase the ultimate caliber of an injured hollow viscus without resort to mechanical bougienage. In addition, the data suggest that wound contraction may play a role in stricture formation in this model.

Structural characterization of calcitonin gene-related peptide purified from rabbit intestine.

Calcitonin gene-related peptide (CGRP) immunoreactive material has been found in extracts of the intestine, however, the structure of intestinal CGRP is not known. Analytical reverse phase HPLC and ion-exchange FPLC revealed one predominant immunoreactive CGRP peak in rabbit intestinal extracts. This material was purified from rabbit intestine by sequential steps of reverse phase HPLC and ion-exchange FPLC. Microsequence and mass spectral analysis of the purified peptide and its chymotryptic fragments were consistent with the structure: GCNTATCVTHRLAGLLSRSGGMVKSNFVPTNVGSEAF-amide. Rabbit intestinal CGRP is identical to human CGRP-II in 35 of 37 amino acid residues. Two amino acid differences were detected at position 1, with Gly in rabbit CGRP instead of Ala in human CGRP-II, and at position 35, with Glu instead of Lys, respectively. Rabbit CGRP differed from human CGRP-I by three additional amino acids at positions 3, 22, and 25. This report shows that a CGRP form which closely resembles human CGRP-II, by means of chemical characterization, is the predominant form in rabbit intestine. Rabbit CGRP is the only CGRP form which has Gly as the amino terminal amino acid. Since the amino terminus of CGRP seems to be important for expression of bioactivity, the biological activity of rabbit CGRP may differ from human, rat and porcine CGRP.

Psychopharmacologic violence associated with cocaine abuse: kindling of a limbic dyscontrol syndrome?

1. An association of cocaine abuse with aggressive or violent behavior arising from direct pharmacologic effects of cocaine is demonstrable in the forensic and clinical literature. 2. The neurobehavioral basis for this association is considered form among known CNS actions of cocaine. A hypothesis is developed concerning the role of pharmacological kindling by cocaine that may sensitize for release of limbic-hypothalamic mechanisms of aggressive behavior, and for a drug-induced dyscontrol syndrome. 3. Parallels are drawn to kindling by electrical stimuli, and to neurophysiological research on mechanisms of aggression. 4. A role of concurrent hyperthermic effects of cocaine is suggested. 5. Potential contributions of cocaine actions on CNS serotonergic, catecholaminergic and/or adenosinergic systems are considered. 6. A likely role of concurrent ethanol ingestion to enhance the manifestation of cocaine-associated violence is recognized. 7. Pharmacological challenges, lidocaine or caffeine, are suggested as a means of detecting lowered thresholds of limbic excitability as a consequence of repeated cocaine exposures.

The effects of cocaine and nandrolone co-administration on aggression in male rats.

1. Cocaine and anabolic-androgenic steroids are among the more commonly abused substances in selected populations. These agents, when used alone or in combination, have been reported to cause aggressive tendencies in both laboratory-based animal models and in human clinical situations. This project, using a resident-intruder paradigm, examined the effects of co-administration of cocaine and a typical anabolic-androgenic steroid, nandrolone decanoate, on the development of aggression in male Sprague-Dawley rats. 2. Dose response studies demonstrated that low dose cocaine (1 mg/kg) produced more aggression in a greater percentage of animals than for either the controls or groups receiving higher doses (up to 20 mg/kg). Initially, high intermittent doses of nandrolone (20 mg twice weekly) produced more aggression; however, low daily doses of nandrolone (2 mg) produced greater levels of aggression following 4 weeks of treatment. 3. Optimal doses of cocaine and nandrolone, when administered together, resulted in aggression scores that were not significantly different from controls or either drug singly. However, a greater percentage of animals receiving both drugs exhibited aggression than did rats receiving either drug alone. 4. These results support the interpretation that the drugs interact to produce unique effects in the development of aggression. However, the complexity and extent of the interactions is great and remains to be fully elucidated.

Premature mortality among prominent American authors noted for alcohol abuse.

An array of 27 American authors noted in the literature for their excessive drinking was compared for their longevity to a control group of 54 writers. The latter were selected for having similar sex distribution and distribution of dates of birth, as well as for their peer recognition among American writers. The median (and range) for the alcohol-abusing writers was 65 (33-77) years, whereas the control group survived to a median of 76 years (range 32-90). This difference was highly significant (p less than 0.001). Thus, data from American authors support those epidemiologic studies reporting alcohol abuse to be a cause of premature mortality, rather than others that have failed to demonstrate such a relationship. The social cost of an alcohol-related decrement in lifespan is emphasized by these data from a highly talented selection of American writers.

Intragastric drug self-administration by rats exposed successively to morphine and ethanol.

Male Sprague-Dawley rats were implanted with intragastric cannulas for self-administration of drug solutions by bar-pressing on a continuous reinforcement schedule in 10-hour daily sessions. Similar levels of responding were observed for doses per infusion of 3.0 mg/kg morphine sulfate and 25 mg/kg ethanol in separate groups of rats. When rats that showed self-administration of a morphine solution over a 5-day period were than given access instead to ethanol for 5 days, the number of infusions taken did not deviate significantly between the two periods. However, rats selected from a small minority that failed to take morphine under these conditions also failed to manifest ethanol self-administration behavior. The data can be seen to support a possible concurrence or similarity between innate factors determining acceptance or rejection of morphine and ethanol as objects of self-administration behavior.

Lethal synergism between morphine or other narcotic analgesics and propranolol.

Interactions of (+/-)-propranolol HCl with various narcotics were determined in albino rats. The 24-h intraperitoneal (i.p.) LD50 of morphine sulfate + saline was 15--16 times greater than for morphine + propranolol in both sexes although morphine was nearly twice as toxic to males as to females. The potency ratios for LD50's with saline vs. with propranolol were: codeine, 1.9, (+/-)-methadone, 6.0; (-)-alpha-acetylmethadol, 2.8 (72 h). The toxicity of levorphanol also was greatly increased with propranolol, but the dose-effect relationship showed non-parallelism vs. levorphanol + saline. Albino mice and mongrel dogs also showed synergism between morphine and propranolol. Mortality after morphine and propranolol was antagonized by naloxone or naltrexone in rats and mice. The potency ratio in rats for morphine + saline vs. morphine + practolol was 3.5. However, the synergism between propranolol and the narcotics probably was unrelated to beta-adrenergic blocking effects of propranolol because of the apparent equivalence of (+)-, (-)- and (+/-)-propranolol in rats for synergism with morphine.

Synergism of the toxicity of physostigmine and neostigmine by lithium or by a reserpine-like agent (Ro4-1284).

A single sublethal i.p. dose of lithium chloride (300 mg/kg or 7.1 meq/kg) followed 12 h later by an otherwise sublethal s.c. dose of physostigmine sulfate (1.0 mg/kg) resulted in 90% mortality among male rats following a pronounced cholinergic syndrome, including convulsions. This confirms a previous report of a lethal synergism of physostigmine after subacute dosing with lithium. Mortality could be completely prevented by 1.0 mg/kg of atropine sulfate given 30 min before physostigmine, but was incompletely, if at all, reduced by selective peripheral cholinergic blockers, methylatropine bromide (0.5, 1.5 mg/kg) or glycopyrrolate (1 mg/kg). This suggested a predominantly central site for the toxic interaction. However, a similar synergism of lethality caused by neostigmine methylsulfate (0.3 mg/kg, s.c.) after treatment with lithium, which could be eliminated by methylatropine or glycopyrrolate, indicates that lithium may also produce lethal synergism of a cholinesterase (ChE) inhibitor that does not act centrally. Ro4-1284, an agent that has reserpine-like actions, was tested in combination with physostigmine or neostigmine; it showed synergism of toxicity nearly the same as in the case of lithium plus the cholinergic agents. These findings support the hypothesis that lithium causes the toxic synergism via a reduction of adrenergic activity, leading to an imbalance between adrenergic and cholinergic influences and a consequent failure to tolerate the effects of the ChE inhibitors. A potential hazard for the clinical use of physostigmine and neostigmine, concurrently with lithium or reserpine-like agents, it suggested.

Toxic interaction between narcotic analgesics and inhibitors of catechol-O-methyltransferase.

A lethal synergism between morphine and tropolone, an inhibitor of catechol-O-methyltransferase, was previously noted in adult male Holtzman rats. The present research demonstrates that this phenomenon generalizes across factors of sex, age, strain (Sprague--Dawley, Wistar) and species (Swiss albino mice). Acute toxicity was also significantly increased (1.5--1.9 times) in the case of codeine, methadone, meperidine and levorphanol, but to a lesser extent than for morphine (4.0 times) in the S-D strain. Another COMT inhibitor, 3,5-dihydroxy-4-methoxybenzoic acid, interacted with morphine in S-D rats to an equal degree as did tropolone. Post-treatment with 1 mg/kg of naloxone in rats or naltrexone in mice reduced the high lethality associated with morphine plus tropolone. There was a pronounced lowering of whole brain norepinephrine (NE) level after morphine plus tropolone in Wistar rats with doses of each component that alone caused no change in NE. Brain dopamine (DA) was elevated by tropolone and by its combination with morphine. Each drug alone caused slight lowering of brain serotonin. Enhancement by tropolone of the toxicity of (+)-amphetamine in mice and rats was of similar magnitude as for morphine. The possible role of brain NE and/or DA in the sensitivity to acute toxic effects of opioids in rodents is suggested by these data, as well as a parallel in this regard with amphetamine-type stimulants.

Dynamic organ culture of precision liver slices for in vitro toxicology.

The lack of a reproducible method for the production of thin tissue slices has hindered the use of liver slices as an in vitro tool for hepatotoxicity studies. Fresh human, rat, and rabbit liver was processed using a mechanical slicer. With this instrument, precision (5% of thickness) liver slices in the submillimeter range could be produced at a rapid rate. Slices were prepared from fresh livers in chilled, oxygenated buffer to minimize trauma. Following incubation for up to 20 h in a dynamic organ culture system, histology of incubated slices suggested that 250 m precision-cut slices were optimum in regard to morphology relative to liver slices incubated under conventional organ culture conditions. Addition of bromobenzene to the culture showed time-dependent hepatotoxicity based on two classic parameters of cell degeneration. Histological evidence is presented which suggests the usefulness of this system for hepatotoxicity studies and the production of focal necrosis in vitro.

Perinatal exposure to cannabichromene and delta 9-tetrahydrocannabinol: separate and combined effects on viability of pups and on male reproductive system at maturity.

The effects of cannabichromene (CBC), delta 9-tetrahydrocannabinol (delta 9-THC) and their combination (all doses 50 mg/kg orally) were determined after being administered to female mice for 7 days beginning on the 20th day of gestation. The THC treatment reduced postnatal viability, impaired male reproductive behavior at maturity and significantly reduced seminal vesicle weights. No changes from control values occurred after CBC or CBC + THC. Thus, CBC alone at this dosage did not act like THC; moreover, it antagonized the effects of THC when the two were given in combination.

Determination of the sedimentary microbial biomass by extractible lipid phosphate.

The measurement of lipid phosphate is proposed as an indicator of microbial biomass in marine and estuarine sediments. This relatively simple assay can be performed on fresh, frozen or frozen-lyophilized sediment samples with chloroform methanol extraction and subsequent phosphate determination. The sedimentary lipid phosphate recovery correlates with the extractible ATP and the rate of DNA synthesis. Pulse-chase experiments show active metabolism of the sedimentary phospholipids. The recovery of added 14C-labeled bacterial lipids from sediments is quantitative. Replicate analyses from a single sediment sample gave a standard deviation of 11%. The lipid extract can be fractionated by relatively simple procedures and the plasmalogen, diacyl phospholipid, phosphonolipid and non-hydrolyzable phospholipid content determined. The relative fatty acid composition can be readily determined by gas-liquid chromatography.The lipid composition can be used to define the microbial community structure. For example, the absence of polyenoic fatty acids indicates minimal contamination with benthic micro-eukaryotes. Therefore the high content of plasmalogen phospholipids in these sediments suggests that the anaerobic prokaryotic Clostridia are found in the aerobic sedimentary horizon. This would require anaerobic microhabitats in the aerated zones.

Evaluation of risperidone in the neonatal 6-hydroxydopamine model of Lesch-Nyhan syndrome.

Rats were treated as neonates with 6-hydroxydopamine (6-OHDA) 100 mg free base in 10 microl intracisternally. Upon maturation, animals were injected with L-dopa and placed in photocell cages for monitoring of locomotion, stereotypies, and self-mutilation. Pretreatment with either risperidone or SCH-23390 significantly reduced locomotion and stereotypies. SCH-23390 eliminated L-dopa induced self-mutilation in all subjects, while risperidone eliminated self-mutilation in all but one subject.