Confirmation of anatoxin-a(s), in the cyanobacterium Anabaena lemmermannii, as the cause of bird kills in Danish lakes

Cyanobacterial blooms were implicated in bird kills at lakes in Denmark in July 1993 and June-July 1994. These blooms were dominated by Anabaena lemmermannii and were shown to contain a neurotoxin with anticholinesterase activity. In this study, the toxin was isolated by mouse lethality guided column chromatographies from the field sample collected at Lake Knud s? in 1993. Various spectroscopic data indicated that the toxin was anatoxin-a(s), an irreversible anticholinesterase, first reported in Anabaena flos-aquae. Chemical detection of the same toxin in cultured A. lemmermannii also confirmed this species as the cause of the deaths of the wild birds.     

Hepatotoxin (microcystin) and neurotoxin (anatoxin-a) contained in natural blooms and strains of cyanobacteria from Japanese freshwaters

Amounts of hepatotoxic microcystin and neurotoxic anatoxin-a were estimated in natural blooms and strains of cyanobacteria from freshwaters in Japan. A simultaneous analysis method of anatoxin-a and microcystin was applied to natural bloom samples, which has been dominated by several species and the strains of cyanobacteria which produced simultaneously both toxins. The natural blooms examined in the present study were mainly composed of Anabaena and Oscillatoria, but most also contained Microcystis and other cyanobacteria. Only one sample was almost unialgal, Anabaena spiroides, collected from Lake Sagami. The toxins in 14 samples collected from nine different natural blooms during 1988-1992 were identified as microcystins-RR, -YR, and -LR; desmethyl-7-microcystin-LR (7-DMLR); and anatoxin-a. Microcystins were the main toxins contained in these natural blooms, with anatoxin-a not being detected or of very little quantity. 7-DMLR was detected in samples only from Lake Kasumigaura. Five strains of Anabaena isolated from waters in Japan produced a small amount of anatoxin-a, but no microcystins. One half of the strains of Microcystis produced microcystins and/or anatoxin-a. This is the first study showing Microcystis producing both anatoxin-a and microcystins.     

First identification of microcystins in Irish lakes aided by a new derivatisation procedure for electrospray mass spectrometric analysis

Recent animal and bird deaths at several lakes in Ireland were indicative of possible cyanobacterial poisoning. Using protein phosphatase inhibition assays, microcystins (MCs) were identified in extracts of cyanobacteria from several lakes at concentrations ranging from 1.6 to 168 micrograms/g. This is the first report of MCs in Irish freshwaters. The protein phosphatase inhibition assay was used to screen fractions during HPLC purification of the MCs in cyanobacteria (Anabaena and Oscillatoria) and water samples from Corbally and Caragh Lakes. MC-LR, MC-HtyR, MC-FR, and MC-YR and 3 unidentified MCs of m/z values 1028.5, 981, 1042.7 were isolated from the Corbally sample; while the Caragh Lake sample contained largely MC-LR and MC-YR. A new microanalytical technique was developed for the confirmation of MCs which involved the derivatisation of the methyldehydroalanine group of MCs with 2-aminoethanethiol. Electrospray mass spectrometry of these products showed characteristic double-charged ions, and this novel technique was useful for differentiating MCs from co-eluting impurities in HPLC fractions of cyanobacterial extracts.     

Two (Z)-dehydrobutyrine-containing microcystins from a hepatotoxic bloom of Oscillatoria agardhii from Soulseat Loch, Scotland

Two (Z)-dehydrobutyrine(Dhb)-containing microcystins, [d-Asp3, (Z)-Dhb7]microcystin-HtyR (1) and [d-Asp3, (Z)-Dhb7]microcystin-LR (2), were isolated from a hepatotoxic bloom of the cyanobacterium Oscillatoria agardhii from a freshwater lake in Scotland. The geometrical structure of the Dhb units in the microcystins was determined as Z on the basis of NOE and ROESY experiments.     

Urinary mercury levels in females: influence of skin-lightening creams and dental amalgam fillings

The influence of application of skin-lightening creams and dental amalgam fillings on the urinary mercury (Hg) level was evaluated in 225 females (ages 17 to 58 years) living in Riyadh, capital of Saudi Arabia. The arithmetic mean of the urinary Hg level was 6.96 +/- 20.43 micrograms 1(-1), in the range 0 to 204.8 micrograms 1(-1). The mean urinary Hg level adjusted by creatinine (Cr) was 11.22 +/- 37.23 micrograms g-1 Cr, in the range 0 to 459.37 micrograms g-1. No significant difference in urinary Hg was noted between the females regarding the use of skin-lightening creams. On the other hand, results showed that urinary Hg concentration was influenced by the use and number of dental amalgam fillings. No women were identified with symptoms or signs that could be attributed to Hg intoxication. Urine analyses for creatinine, urea, uric acid, phosphorus, magnesium, glucose and calcium showed significant correlation with urinary Hg. This suggests that chronic exposure to Hg may be associated with a deterioration of renal function.     

Postnatal changes in size and actomyosin content of rabbit gastric myocytes

Maximal tension generated by gastric muscle is three to four times greater in weanlings than in newborn rabbits. To determine if this functional maturation is accompanied by structural changes, we compared length-tension relationships, myocyte number and size, and actomyosin content in muscle from the gastric body of newborn (1 day) and weanling (12 weeks) rabbits. Passive tension at optimal length (Lo) was six times greater in circular smooth muscle strips from weanling rabbits than from newborn rabbits. Active tension at Lo in weanling rabbits was three times greater than in newborn rabbits. For morphometry, muscle cross-sections stretched to Lo in the circular axis were photographed with electron microscopy (5300x). Cell number/unit area was counted in circular muscle layers from newborns and weanlings. Cross-sectional area of each cell was measured by computerized planimetry. There were 2.5 times more cells per unit area in newborn than in weanling tissue, P < 0.001. However, the mean cell area in newborns (5.4 +/- 4.6 microns2) was less than that in weanlings (13.5 +/- 11.7 microns2). Consequently, the muscle cells occupied similar total areas in newborns and weanlings. We measured actin and myosin heavy chain in full-thickness muscle homogenates using SDS gel electrophoresis and densitometric scanning. Actin and myosin concentrations were lower in newborns (9.6 +/- 1.3 micrograms g-1 wet weight and 5.6 +/- 0.7 micrograms g-1 wet wt, respectively) than in weanlings (17.7 +/- 3.0 micrograms g-1 wet wt and 8.2 +/- 1.6 micrograms g-1 wet wt respectively), each P < 0.01. The proportion of myosin heavy chain isozymes did not change with age. We conclude that there are postnatal increases in cell size and the quantity of actin and myosin in rabbit gastric muscle. The increase in quantity of contractile protein may be in part responsible for age-dependent increases in maximal tension.     

Peroxynitrite formation in focal cerebral ischemia-reperfusion in rats occurs predominantly in the peri-infarct region

The plasma pharmacokinetics of danofloxacin administered at 1.25 mg kg-1 body weight by the intravenous and intramuscular routes were determined in sheep. Tissue distribution was also determined following administration by the intramuscular route at 1.25 mg kg-1 body weight. Danofloxacin had a large volume of distribution at steady state (Vdss) of 2.76 +/- 0.16 h (mean +/- S.E.M.) L kg-1, an elimination half-life (t1/2 beta) of 3.35 +/- 0.23 h, and a body clearance (C1) of 0.63 +/- 0.04 L kg-1 h-1. Following intramuscular administration it achieved a maximum concentration (Cmax) of 0.32 +/- 0.02 microgram mL-1 at 1.23 +/- 0.34 h (tmax) and had a mean residence time (MRT) of 5.45 +/- 0.19 h. Danofloxacin had an absolute bioavailability (F) of 95.71 +/- 4.41% and a mean absorption time (MAT) of 0.81 +/- 0.20 h following intramuscular administration. Mean plasma concentrations of > 0.06 microgram mL-1 were maintained for more than 8 h following intravenous and intramuscular administration. Following intramuscular administration highest concentrations were measured in plasma (0.43 +/- 0.04 microgram mL-1), lung (1.51 +/- 0.18 micrograms g-1), and interdigital skin (0.64 +/- 0.18 microgram g-1) at 1 h, duodenal contents (0.81 +/- 0.40 microgram mL-1), lymph nodes (4.61 +/- 0.35 micrograms g-1), and brain (0.06 +/- 0.00 microgram mL-1) at 2 h, jejunal (10.50 +/- 4.31 micrograms mL-1) and ileal (5.25 +/- 1.67 micrograms mL-1) contents at 4 h, and colonic contents (8.94 +/- 0.65 micrograms mL-1) at 8 h.     

Herbicide residues in leaves of Erythroxylum coca var. coca plants treated with soil-applied tebuthiuron and hexazinone

The herbicide residue levels in leaves of Erythroxylum coca var. coca Lam. plants treated with soil applications of tebuthiuron and hexazinone at 3.36 and 6.72 kg a.i. ha-1 were determined in order to estimate the potential for human exposure to these residues from consuming the leaves or cocaine produced from them. Field-grown plants were treated with a commercial formulation of tebuthiuron or hexazinone and leaves were harvested at the first indication of herbicide injury (i.e. chlorosis and/or necrosis) and at the onset of leaf abscission. Herbicide residues were detected by HPLC in leaf samples from both harvests of all plants treated with tebuthiuron or hexazinone. At 3.36 kg ha-1, herbicide residues in the leaves were less than 2 micrograms g-1 dry wt. for both harvests of both experiments. The highest residue levels detected were 5.90 micrograms g-1 dry wt. for tebuthiuron and 7.17 micrograms g-1 dry wt. for hexazinone in leaves from plants treated with the herbicide at the rate of 6.72 kg ha-1 and harvested at the onset of leaf drop. Based on published toxicity data and estimates of leaf consumption, the herbicide residues in leaves of E. coca var. coca plants treated with tebuthiuron or hexazinone at twice the recommended control rates or less would have a negligible contribution to the health risks of individuals who chew coca leaves. Furthermore, based on the most conservative estimates of cocaine yield and herbicide carry over, death by cocaine overdose would occur long before the NOEL for either herbicide was reached.     

Quantitation of acetazolamide in rat plasma, brain tissue and cerebrospinal fluid by high-performance liquid chromatography

A simple and sensitive high-performance liquid chromatographic method for the analysis of acetazolamide (AZ) in rat blood (plasma/serum, whole blood and serum ultrafiltrate), brain tissue and cerebrospinal fluid (CSF) was described. Quantitative extraction of AZ with ethyl acetate from both buffered plasma and brain tissue homogenate (pH 8.0) was achieved. Each extract was evaporated to dryness and the residue was chromatographed on a reversed-phase column. CSF was directly analysed without extraction step. The limits of detection were 0.05 microgram ml-1 for plasma, 0.02 microgram g-1 for brain tissue and 0.004 microgram ml-1 for CSF. Calibration curves were linear over the working ranges of 0.1-100 micrograms ml-1 for plasma, 0.05-50 micrograms g-1 for brain tissue and 0.025-50 micrograms ml-1 for CSF. The reproducibility of AZ assay in the rat biologic media indicated very low relative standard deviations (RSDs). The recoveries of AZ added to plasma and brain tissue were more than 96% with an RSD of less than 5%. The present method was applied to studies of plasma concentration profiles of the drug after administration and its distribution into central nervous system.     

Effect of systemic corticoid and antihistamine alone or in combination on elements of the response to a two dose nasal allergen challenge

Adenosine, an endogenous vasodilator, induces a cerebral vasodilation at hypotensive infusion rates in anaesthetized humans. At lower doses (< 100 micrograms kg-1 min-1), adenosine has shown to have an analgesic effect. This study was undertaken to investigate whether a low dose, causing tolerable symptoms of peripheral vasodilation affects the global cerebral blood flow (CBF). In nine healthy volunteers CBF measurements were made using axial magnetic resonance (MR) phase images of the internal carotid and vertebral arteries at the level of C2-3. Quantitative assessment of CBF was also obtained with positron emission tomography (PET) technique, using intravenous bolus [15O]butanol as tracer in four of the subject at another occasion. During normoventilation (5.4 +/- 0.2 kPa, mean +/- s.e.m.), the cerebral blood flow measured by magnetic resonance imaging technique, as the sum of the flows in both carotid and vertebral arteries, was 863 +/- 66 mL min-1, equivalent to about 64 +/- 5 mL 100 g-1 min-1. The cerebral blood flow measured by positron emission tomography technique, was 59 +/- 4 mL 100 g-1 min-1. All subjects had a normal CO2 reactivity. When adenosine was infused (84 +/- 7 micrograms kg-1 min-1.) the cerebral blood flow, measured by magnetic resonance imaging was 60 +/- 5 mL 100 g-1 min-1. The end tidal CO2 level was slightly lower (0.2 +/- 0.1 kPa) during adenosine infusion than during normoventilation. In the subgroup there was no difference in cerebral blood flow as measured by magnetic resonance imaging or positron emission tomography. In conclusion, adenosine infusion at tolerable doses in healthy volunteers does not affect global cerebral blood flow in unanaesthetized humans.     

Simulation of CA3 region of hippocampus by kinetic models

Embryos of grayling (Thymallus thymallus) were exposed to different concentrations of methylmercury (0.16, 0.8, 4.0 and 20 micrograms Hg l-1) during the first 10 days of development. The exposure resulted in body concentrations in the newly hatched fry of 0.09, 0.27, 0.63 and 3.80 micrograms Hg g-1 wet wt., respectively. A control group had a body concentration of 0.01 microgram Hg g-1. Morphological disturbances were only found in the highest exposure group. Three years later, at a size of 13.8 +/- 0.8 cm, the different groups were tested for sublethal toxicant effects on foraging behavior. In the first series of experiments we tested the foraging efficiency of the fish when kept alone for 5 min in small flow-through aquariums. In the second series of experiments we tested the competitive ability of eight individuals from an exposed group vs. eight individuals from a control group when kept together for 30 min in a 300-1 aquarium. In both experiments live Dapnia magna were used as prey. We found impaired feeding efficiencies and reduced competitive abilities in grayling from the exposed groups which as yolk-fry had Hg concentrations of 0.27 microgram g-1 or more. In the foraging efficiency experiments these groups were 15-24% less efficient as compared to the control group. In the competitive ability experiments the control group caught two to six times as many preys as these exposed groups. Such harmful body concentrations of Hg (> 0.27 microgram g-1) may be found in eggs from piscivorous fishes in lakes receiving diffuse atmospheric depositions of mercury. We suggest such concentrations may have ecological consequences by reducing the fitness of the affected populations.     

Sheep mortality associated with paralytic shellfish poisons from the cyanobacterium Anabaena circinalis

This is the first report of sheep mortalities associated with paralytic shellfish poisons (PSPs) from the cyanobacterium Anabaena circinalis Rabenhorst. Fourteen sheep died within 150 m of a farm dam containing a dense bloom of A. circinalis. Extracts from both the cyanobacterium and small intestine from a dead ewe were analysed by high-performance liquid chromtography (HPLC) and found to contain PSPs. The toxin profiles of the cyanobacterium contained a high proportion of C-toxins (70%), whereas toxins in the small intestine content were dominated by gonyautoxin 5 (87%). This observation could be explained by desulfation of the C-toxins in the gut of the sheep. The LD100 of bloom material calculated from HPLC data was consistent with mouse bioassay data (12-25 mg/kg). The symptoms of affected sheep, mouse bioassay data, coupled with HPLC analysis of toxins from the bloom samples and the intestine contents, and the absence of other cyanobacterial alkaloid toxins, indicate that PSPs were responsible for the deaths of the sheep.     

Exposure to methyl tertiary-butyl ether from oxygenated gasoline in Stamford, Connecticut

In 1993, state health officials in Connecticut invited the Centers for Disease Control and Prevention (CDC) to assist in an investigation of exposure to methyl tertiary-butyl ether in oxygenated gasoline in Stamford, Connecticut. Venous blood samples were collected from 14 commuters and from 30 other persons who worked in the vicinity of traffic or automobiles, and the samples were analyzed for methyl tertiary-butyl ether, tertiary-butyl alcohol, benzene, m-/p-xylene, o-xylene, and toluene. The highest levels of methyl tertiary-butyl ether in blood were measured among gasoline service station attendants (median = 15 micrograms/l, range = 7.6-28.9 micrograms/l). Blood levels of methyl tertiary-butyl ether were highly variable among persons who worked in car-repair shops (median = 1.73 micrograms/l, range = 0.17-36.7 micrograms/l) and were generally lowest among commuters (median = 0.11 micrograms/l, range = < 0.05-2.60 micrograms/l). Blood levels of methyl tertiary-butyl ether were correlated strongly with personal-breathing-zone samples of methyl tertiary-butyl ether and blood levels of other volatile organic compounds. This exposure information should prove useful to a future risk analysis of this high-volume chemical.     

Percutaneous absorption and disposition studies of methotrexate in rabbits and rats

The absorption and disposition of methotrexate (MTX) in the plasma, synovial fluid (SF), skin, and muscle tissue were studied following administration of a topical MTX gel in rabbits and rats. In rabbits, MTX concentrations in the plasma increased steadily toward the peak (5.9 +/- 2.8 ng mL-1) which appeared at approximately 2 h postdose and declined with the elimination half-life of 4.48 +/- 1.74 h. At 1 h after the topical dose, the MTX concentrations in the skin (49.0 +/- 19.8 micrograms g-1), muscle (12.7 +/- 3.3 ng g-1), and SF (19.2 +/- 10.1 ng g-1) underneath the dosed stifle joint were significantly higher (p < 0.05) than those of the untreated stifle joint, indicating the potential therapeutic value of topical delivery of MTX for rheumatoid arthritis. A large fraction (approximately 59%) of MTX which was found in the skin at 1 h postdose was present in the stratum corneum, indicating its extensive binding capacity for MTX. The MTX concentrations in the muscle and SF of the dosed stifle joint at 1 h postdose were 1.8 and 2.6 times higher than those in the dosed elbow joint, respectively, reflecting the effect of dose site on the permeation of MTX. Using a new filter paper method, the amounts of SF obtained from the elbow and stifle joints of four rabbits were 26.3 +/- 8.3 and 48.8 +/- 5.2 mg, respectively. A significant enhancer effect of N,N-diethyl-n-toluamide (DEET) on the disposition of MTX in the stratum corneum of rabbit ear was observed (p < 0.05) by the tape-stripping method. In rats, the gel containing 4% DEET resulted in a twofold increase in the permeation of MTX into the muscle over the 4 h period postdose. A modified HPLC method with a linear calibration curve (r > 0.999) over the range of 2-50 ng mL-1. quantitation limit of 0.5 ng mL-1, and mean recovery of approximately 87% was used for the quantitation of MTX in the tissue and fluid samples.     

Cerebral blood flow velocity in relation to cerebral blood flow, cerebral metabolic rate for oxygen, and electroencephalogram analysis during isoflurane anesthesia in dogs

The purpose of this study was to correlate changes in cerebral blood flow velocity (Vmean) with cerebral blood flow (CBF) during isoflurane anesthesia in dogs. The relation between cerebral oxygen consumption (CMRO2) and electroencephalogram (EEG) analysis also was investigated. Blood flow velocity was measured in the middle cerebral artery using a pulsed transcranial Doppler (TCD). CBF was measured with radioactive microspheres. EEG was measured over both hemispheres and median EEG frequency (median frequency) was calculated after fast Fourier transformation. Baseline anesthesia was maintained with 50% nitrous oxide in oxygen and 50 micrograms.kg-1 x h-1 fentanyl. Animals of Group I (control, n = 6) were not given isoflurane. Data were recorded at baseline, and at 30, 60, and 90 min. There was no significant change in any variable over time. In Group II (n = 7), data were recorded at baseline and at 1%, 2%, and 3% end-tidal isoflurane. Mean arterial pressure was maintained at baseline levels by phenylephrine infusion. CBF increased from 70.8 +/- 10.6 mL.100g-1 x min-1 at baseline to 146.1 +/- 36.9 mL.100 g-1 x min-1 with 3% isoflurane (P < 0.01). Vmean increased from 38.3 +/- 6.7 cm/s to 65.6 +/- 9.7 cm/s (P < 0.01). The correlation between relative changes in CBF and Vmean was r = 0.94 (P < 0.01). With 1% isoflurane the EEG shifted to slow-wave, high-voltage activity, and median frequency decreased from 5.9 +/- 0.7 Hz to 1.4 +/- 0.4 Hz (P < 0.05). Median frequency was not decreased further during 2% and 3% isoflurane anesthesia.(ABSTRACT TRUNCATED AT 250 WORDS)     

The disposition of nifurtimox in the rat isolated perfused liver: effect of dose size

The disposition of nifurtimox was studied in the rat isolated perfused liver using a recirculating system. The drug was administered as a bolus (5.0, 15.0 or 30.0 micrograms mL-1), and its disappearance was monitored by analysing perfusate samples. In all experiments perfusate disappearance was monoexponential, and no significant difference was found between the three doses for the elimination constant (0.016, 0.011 and 0.012 min-1, respectively), half-life (46.6, 65.8 and 66.8 min, respectively), extraction rate (0.128, 0.091 and 0.099, respectively) and distribution volume (41.1, 47.3 and 30.7 mL g-1, respectively). At 30 micrograms mL-1 the hepatic clearance was lower than the other concentrations of nifurtimox (0.66, 0.51 and 0.34 mL min-1 g-1, respectively). Relatively little parent drug was recovered from the liver at the end of the perfusions. In summary, nifurtimox is cleared slowly from the rat isolated perfused liver, is poorly extracted by hepatocyte cells and is completely metabolized from 2 to 4 h after perfusion.     

Chemical modulation of in situ intrinsic cardiac neurones influences myocardial blood flow in the anaesthetised dog

OBJECTIVE: The aim was to determine whether modulation of intrinsic cardiac neurones influences the distribution of myocardial blood flow in canine anaesthetised open chest experimental preparations. METHODS: Intrinsic cardiac neurones were modified by locally applied nicotine (100 micrograms) or bradykinin (50 micrograms) while changes were recorded in cardiac haemodynamics and myocardial blood flow (radiolabelled microspheres). Right and left ventricular intramyocardial tissue pressures were measured with high fidelity microtip transducers. RESULTS: Control injections of saline (vehicle; 0.1 ml) into active loci did not produce cardiovascular responses. Nicotine modulation of intrinsic cardiac neurones did not change coronary artery conductance, but total myocardial blood flow [116(SEM 17) v 532(97) ml.min-1.100 g-1; p = 0.001 v baseline] and oxygen consumption [7.92(1.10) v 20.14(1.86) ml.min-1.100 g-1; p = 0.001] increased in direct relation to heart rate-blood pressure product changes. Locally administered bradykinin increased coronary artery conductance [2.62(0.39) v 4.71(1.07) ml.min-1.100 g-1.mm Hg-1], total myocardial blood flow, to 263(72) ml.min-1.100 g-1, and oxygen consumption, to 14.9(4.4) ml.min-1.100 g-1; however, heart rate-blood pressure product did not change. CONCLUSIONS: These results support earlier findings that intrinsic neurones are involved in cardiac regulation. Furthermore, modification of intrinsic cardiac neurones by nicotine or bradykinin significantly alters the distribution of myocardial blood flow, possibly because of increased myocardial metabolism.     

Evaluation of carborane-containing porphyrins as tumour targeting agents for boron neutron capture therapy

A number of carborane-containing porphyrins were administered to mice bearing subcutaneously transplanted mammary carcinomas. Administration was via serial intraperitoneal (i.p.) injections to assess their relative toxicities and tumour affinities. Three analogues of the natural porphyrin heme and four tetraphenylporphyrins (TPPs) were given at total doses of 78-245 micrograms g-1 body weight. The water-insoluble TPPs were less toxic to mice, and delivered greater amounts of boron to tumour than did the water-soluble TPPS and the heme analogues. One such compound, NiTCP-H, delivered more than 100 micrograms B g-1 to tumour tissue with a tumour:blood boron concentration ratio greater than 500:1 and a tumour: brain boron concentration ratio greater than 50:1, 4 days after the last of six i.p. injections given over 2 days. Another TPP analogue, NiTCP, delivered approximately 50 micrograms B g-1 to tumour with similar boron concentrations in normal tissues. Neither compound was toxic to mice at total doses of approximately 200 micrograms g-1 body weight. In contrast, the heme analogues were toxic and, with the exception of VCDP, delivered less boron to tumour than NiTCP and NiTCP-H. The two porphyrins with the greatest potential for application to boron neutron capture therapy (BNCT), NiTCP and NiTCP-H, yielded higher tumour:blood and tumour:brain boron concentration ratios in mice than could be achieved with p-boronophenylalanine (BPA) and sodium mercaptoundecahydrododecaborate (BSH), the compounds which are currently being used in clinical trials of BNCT in the treatment of glioblastoma. The boron delivered by each of the porphyrins tested remained in tumour tissue longer than did boron delivered by either BPA or BSH. The copper and nickel chelates of these porphyrins behave identically in vivo. The former offer the potential for imaging by 67Cu-mediated single photon emission computed tomography (SPECT) to aid BNCT treatment planning.     

Determination of iodine in milk and oyster tissue samples using combustion and peroxydisulfate oxidation

Two methods are described for the preparation of samples for total iodine measurement in biological matrices. In the first method, the samples were combusted in a stream of oxygen to release iodine that, subsequently, was trapped in a solution as iodide. The second method is a new approach in which the samples were oxidized in a basic solution of peroxydisulfate. In this case, the iodine was retained in solution as iodate. Total iodine was measured by gas chromatographic analysis of the 2-iodopentan-3-one derivative. The methods were tested using Standard Reference Materials (SRMs) 1549 Non-Fat Milk Powder, and 1566a and 1566 Oyster Tissue. Also, KI and KIO3 were used for testing the procedures. The results obtained for the SRMs, given as average +/- standard deviation in micrograms g-1, were: 3.39 +/- 0.14 and 3.40 +/- 0.23 for SRM 1549; 4.60 +/- 0.42 and 4.51 +/- 0.45 for SRM 1566a; and 2.84 +/- 0.16 and 2.76 +/- 0.06 for SRM 1566; values corresponding to combustion and wet oxidation, respectively. Overall, the absolute recoveries varied between 91 and 103%. These methods can also be used in the preparation of targets for the measurement of 129I using accelerator mass spectrometry.     

Nervous regulation of the tone of the retractor penis muscle in the goat

The nervous control of the retractor penis muscle (rp) was investigated in the anaesthetized goat. Also, isolated field stimulated strips of the muscle were studied. The noradrenaline (NA) and acetylcholine (ACh) content of the rp was determined, and histochemistry for adrenergic and acetylcholinesterase (AChE) positive nerves was performed. The muscle exhibited spontaneous activity that persisted after section of all nerves. There was, however, also a tendency of the activity to follow the general vasomotor tone, which disappeared after section of the sympathetic chains. The excitatory adrenergic nerves which innervate the muscle come from the sympathetic chains and run along the pudendal, the hypogastric and the pelvic nerves. The rp has a dense network of adrenergic fibres and is very sensitive to excitatory adrenergic stimulation. It has a fairly large NA content, which is higher in old goats (5.95 +/- 0.42 micrograms g-1) than in young goats (2.87 +/- 0.78 micrograms g-1). Inhibitory non-adrenergic non-cholinergic (NANC) innervation reaches it via the pelvic and the hypogastric nerves. The maximum inhibitory response is reached at low frequencies (2-4 Hz). Cholinergic prejunctional inhibition of the excitatory response to sympathetic chain stimulation was effected by simultaneous stimulation of the hypogastric nerves. In vitro experiments confirmed the presence of endogenous cholinergic muscarinic suppression of the excitatory adrenergic neurotransmission. Significant amounts of ACh (0.81 +/- 0.18 micrograms g-1) are present in the muscle, and it contains strongly AChE positive nerve fibres and nerve cell bodies. It is concluded that the goat rp is innervated by sympathetic adrenergic excitatory nerves and parasympathetic NANC inhibitory nerves. It further has a direct sympathetic inhibitory NANC innervation, and an indirect inhibitory cholinergic innervation which at least in part is sympathetic.