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Kinetics of elimination of thiocyanate in 7 healthy subjects and in 8 subjects with renal failure.

The concentration of thiocyanate in the serum of eight test subjects with renal failure and seven healthy control subjects was measured, as it declined with time, after oral doses of thiocyanate or i.v. injections of nitroprusside had been administered. Additional measurements were taken, on the healthy subjects only, of the concentrations of thiocyanate in the urine, and also of the influence of an increased chloride intake on the rate of elimination of thiocyanate. For the healthy subjects an elimination half-life of between one and five days (mean c. 3 days) was found. Increasing the chloride elimination rate to approximately twice normal did not significantly speed up the rate of thiocyanate elimination. The amounts of thiocyanate which had been administered as doses reappeared almost exclusively in the urine. For the subjects with renal failure, the elimination half-life had a mean value of approximately nine days. The elimination constants were found to be proportional to the creatinine-clearance rates. The ke value at a creatinine-clearance of zero ml/min was approximately 15% of the ke value at a creatinine-clearance rate of 120 ml/min. The distribution volumes for thiocyanate were greater for the patients with renal failure than for the healthy subjects. The conclusions for therapies using nitroprusside are discussed.

Adult

Effect of thiocyanate on nitrite estimation and the cleavage of nitrosamines.

Thiocyanate or bromide increased the colour formed by nitrite reacting with sulfanilic acid and naphthylethylenediamine. If the colour reagents were added together with thiocyanate (final concentration, M/10), the colour intensity was doubled. If sulfanilic acid was added three minutes before addition of naphthylethylenediamine, the relationship between nitrite concentration and colour production was more linear in the presence of thiocyanate. This effect was due to thiocyanate catalysing the diazotization of sulfanilic acid and inhibiting the reaction of nitrite with naphthylethylenediamine. Bromide and thiocyanate are similar in their catalytic effects on nitrosation, and hydrobromic acid in glacial acetic acid is an effective reagent for denitrosation of nitrosamine. Although thiocyanate catalysed denitrosation of nitrosamines, the effect was small except with nitrosomethylaniline, which had also been found to be denitrosated by sulfanilic acid. Thiocyanate could not be used generally for the destruction of nitrosamines; it was also found to be ineffective as an alternative to hydrobromic acid in the estimation of nitrosamines.

Acetates

Smoking during pregnancy--effects on the fetus and on thiocyanate levels in mother and baby.

Twenty-eight mothers smoking 10--20 cigarettes daily during pregnancy had significantly higher serum thiocyanate concentrations at delivery compared in 25 non-smoking controls. The thiocyanate levels were positively correlated to cigarette consumption and inversely correlated to the birth weights of the infants. A highly significant correlation existed between serum thiocyanate levels of the mother and umbilical cord serum thiocyanate levels, reflecting a nearly complete equilibration. The thiocyanate concentrations in human milk on the 4th day after delivery were considerably lower than the serum concentrations, and no correlation existed between serum and milk concentrations. The infants of smoking mothers had significantly decreased weight and length at birth compared in infants of non-smokers. Birth weights were 3 344 +/- 434 g and 3 620 +/- 504 g respectively (p less than 0.05), and lengths 49.8 +/- 1.7 cm and 51 +/- 1.6 cm respectively (p less than 0.05). No differences were found between smokers and non-smokers in placental and umbilical cord histology, and umbilical cord artery medial area. It is concluded that serum thiocyanate concentration in smokers may be used as an objective measure for smoke exposure, and that maternal cigarette smoking acts as an exogenous factor which interferes with intrauterine development of the fetus in a dose related way.

Birth Weight

Method for determining thiocyanate in serum and urine.

We describe a method for rapid and specific measurement of thiocyanate in serum or urine. We separate thiocyanate from interfering compounds by adsorbing it on an anion-exchange resin that has special affinity for thiocyanate, then eluting with sodium perchlorate. The eluted thiocyanate is quantified by a modified König reaction, sodium hypochlorite being used as the chlorinating reagent. Analytical recovery of thiocyanate added to serum and urine was quantitative; the coefficient of variation was 2.3% for both within-day and between-day precision. Cyanide and certain antibiotics interfere, but may be eliminated by including additional washing steps in the usual procedure. The proposed procedure was compared with another method, based on the oxidation of thiocyanate to cyanide. Agreement was satisfactory, both for serum and urine.

Chromatography, Ion Exchange

Effect of cyanide antidotes on the metabolic conversion of cyanide to thiocyanate.

The excretion of thiocyanate following the administration of equitoxic doses of cyanide to unprotected mice and to animals pretreated with various cyanide antidotes has been studied. The results demonstrate that cyanide given alone or to animals pretreated with thiosulfate is extensively converted to thiocyanate. Animals pretreated with sodium nitrite or a combination of nitrite and sodium thiosulfate excreted even higher amounts of thiocyanate. This demonstrates that cyanide originally detoxified by combination with methemoglobin is ultimately converted to thiocyanate in the animal body. Pretreatment of animals with cobalt compounds (cobaltous chloride or dicobalt-EDTA) or a combination of cobalt compounds and thiosulfate resulted, on the other hand, in a less efficient conversion of cyanide to thiocyanate. The cyanide detoxified by trapping as highly undissociated cobalt-cyanide complexes is instead excreted in the urine, as demonstrated by detection of high amounts of cobalt ions and strongly complex-bound cyanide in the urine from animals treated with cobalt compounds and cyanide. A method for the determination of cyanide present as cobalt-cyanide complexes is described and its forensic application is proposed.

Animals

Effects of prolonged cyanide and thiocyanate feeding in rats.

Male weaning rats were fed a semipurified diet containing 10% casein; this diet restricted in methionine, vitamin B12, and iodine; or the complete and restricted diets supplemented with either 1500 ppm potassium cyanide or 2240 ppm potassium thiocyanate for 11.5 mo. There were no deaths or clinical signs of toxicity. Cyanide, but not thiocyanate, caused a consistent reduction in weight gain in the complete and restricted groups. Both cyanide and thiocyanate caused decreased thyroid gland activity in young rats, particularly in the restricted groups. Plasma thyroxine concentrations were maintained in the mature cyanide-treated rats, even though secretion rates were decreased. However, the mature thiocyanate-treated animals showed decreased plasma thyroxine concentrations, despite thyroid gland enlargement. Modest primary myelin degeneration in the spinal cord white matter was found in the restricted group and in rats receiving this diet supplemented with either cyanide or thiocyanate. The lesions did not resemble those of a vitamin B12 deficiency in appearance or those of acute cyanide intoxication in distribution. Because of tissue autolysis, it could not be determined whether these changes resulted from histotoxic anoxia or an alteration of oligodendroglial myelin metabolism.

Animals

Chronic administration of cyanide: urinary excretion of thiocyanate in male and female rats.

The urinary elimination of thiocyanate was investigated in male and female rats following the chronic administration of potassium cyanide. Female rats dosed at the level of 5 mg KCN/kg once a week and twice a week respectively, displayed no significant difference in the excretion of thiocyanate in urine after periods of up to eight weeks of study. Similarly, male rats that were administered 5 mg/KCN/kg twice weekly showed no significant difference in the amount of thiocyanate excreted. The elimination patterns of thiocyanate in male and female rats showed no significant differences. The results suggest that a substrate saturation phenomenon is not operative with cyanide metabolism at the dosage level of potassium cyanide employed in this study. This is contrary to a previously published study which reported that the urinary excretion of thiocyanate decreased after chronic potassium cyanide administration.

Animals

Nitrite and thiocyanate in the fasting and secreting stomach and in saliva.

The concentrations of nitrite and thiocyanate in fasting and pentagastrin stimulated gastric juice and in saliva have been examined. Nitrite was found in all of 17 samples of fasting gastric juice, mean 4-9 +/- 1-1 muM. Stimulation of gastric secretion with pentagastrin caused no significant change in nitrite concentration. Thiocyanate was detected in all of 21 samples of fasting gastric juice and the difference in concentration between smokers and non-smokers probably reflects similar differences in saliva. In contrast to the nitrite data there was a significant drop in thiocyanate concentration of gastric juice after pentagastrin from 0-9 +/- 0-1 mM to 0-3 +/- 0-04 mM, suggesting a salivary origin for the thiocyanate in gastric juice. Thiocyanate is a powerful catalyst of nitrosation, which, together with small amounts of nitrite and naturally occurring amines could lead to the intragastric formation of carcinogenic nitrosamines and in certain circumstances be a factor in the aetiology of gastric cancer.

Fasting

Characteristics of Thiobacillus thioparus and its thiocyanate assimilation.

Thiocyanate-assimilatig bacterium, TK 21, was isolated from activated sludge used for the treatment of thiocyanate contained in coke-oven liquor. This organism oxidized thiosulfate and elemental sulfur, causing a decrease of pH of the medium. These facts indicated that it belongs to the genus Thiobacillus. Potassium thiocyanate (0.5 g/l) was completely assimilated during 60 h. Thiosulfate inhibited the assimilation of thiocyanate but elemental sulfur did not. This bacterium did not evolve cyanide as its oxidation product after the decomposition of thiocyanate. The isoalted bacterium was identified as Thiobacillus thioparus. Examination of the composition of cellular fatty acid of three strains of T. thioparus showed that they prossessed 3-hydroxy fatty acid of C10 and C12; saturated straight chains of C10, C12, C15, C16, C17, and C18; monounsaturated straight chains of C16 and C18; and cyclopropane acid of C17.

Fatty Acids

Thiocyanate inhibition of ATPase and its relationship to anion transport.

Adenosine triphosphatase activity not dependent on sodium or potassium but inhibited by thiocyanate is present in broken-cell homogenates of eel gill and rat kidney. This enzymatic property is predominantly associated with mitochondria, although thiocyanate-inhibited ATPase can also be detected in microsomes with little or no mitochondrial contamination as measured by the activity of the mitochondrial marker enzyme succinic dehydrogenase. When eels are transferred from fresh to salf water, thus increasing active outward transport of chloride across the gill, the thiocyanate-inhibited ATPase of gill microsomes does not change, though the activities of succinic dehydrogenase and Na-K-ATPase in gill homogenates are augmented. The thiocyanate-inhibited ATPase of homogenates of outer renal medulla does not differ from that of renal cortex, in contrast to Na-k-atpase which is higher in renal medulla than in cortex. The data do not support a role for thiocyanate-inhibited ATPase in active chloride transport by epithelial tissues.

Adenosine Triphosphatases

Use of thiocyanic acid to form 2-thiohydantoins at the carboxyl terminus of proteins.

The chemistry of the formation of 2-thiohydantoins on the carboxyl terminal of peptides or proteins was investigated. It was found that thiocyanic acid was much more reactive for the formation of 2-thiohydantoins than were the thiocyanate salts. The physical reasons for this observation are explained. The kinetics of the reaction of a number of proteins, and some of their fragments, with thiocyanic acid were also determined. Simple and safe procedures for the preparation of anhydrous thiocyanic acid solutions were devised. The prospective application of these procedures to sequencing from the carboxyl terminal of a polypeptide is discussed.

Amines

Lactoperoxidase, peroxide, thiocyanate antimicrobial system: correlation of sulfhydryl oxidation with antimicrobial action.

The antimicrobial activity of the lactoperoxidase, peroxide, thiocyanate system against Escherichia coli was directly related to the oxidation of bacterial sulfhydryls. Lactoperoxidase catalyzed the oxidation of thiocyanate, which resulted in the accumulation of hypothiocyanite ion, OSCN-. A portion of the bacterial sulfhydryls were oxidized by OSCN- to yield sulfenic acid and sulfenyl thiocyanate derivatives. The remaining sulfhydryls were not oxidized, although OSCN- was present in large excess. The oxidation of sulfhydryls to sulfenyl derivatives inhibited bacterial respiration. This inhibition could be reversed by adding sulfhydryl compounds to reduce the sulfenyl derivatives and the excess OSCN-. Also, this inhibition could be reversed by washing the cells so as to remove the excess unreacted OSCN-. After washing, the bacteria underwent a time-dependent recovery of their sulfhydryl content. This recovery resulted in recovery of the ability to respire. The inhibited cells were viable if diluted and plated shortly after the incubation with the lactoperoxidase, peroxide, thiocyanate system. On the other hand, long-term incubation in the presence of the excess OSCN- resulted in loss of viability. Also, the inhibition of respiration became irreversible. During this long-term incubation, the excess OSCN- was consumed and the sulfenyl derivatives disappeared.

Dithiothreitol

Correlation between per cent discharge of thiocyanate test and thyroid functions in patients with Hashimoto's disease.

The thiocyanate test was carried out in 18 patients with Hashimoto's disease. Thiocyanate was given orally several hours after the administration of tracer doses of 131I when the thyroid counts seemed to have reached a plateau. The per cent discharge of thiocyanate test was significantly higher in the groups with lower level of T4 (less than 5.5 microgram/100 ml), RSH (less than 25.5%) and BMR (less than +2.5%), and higher levels of TSH (greater than 10.0 muU/ml). A chi-square (chi2) test with Yates correction led to chi2 = 3.56 (1 df) with P = 0.06 (per cent discharge vs T4, RSU, BMR and TSH). Then the per cent thiocyanate discharge was increased with the degree of hypothyroidism in patients with Hashimoto's disease. The correlation between the per cent discharge and 131I 24 h uptake was not significant. It was apparent that the iodide concentration mechanism was present even in the severe hypothyroid stage of Hashimoto's disease. This disproportion between the uptake of iodide and the iodide organification may result in the increase in unbound iodide. It is concluded that the hypothyroidism in Hashimoto's disease may not be caused by a defect in iodide organification, even if it correlated to the degree of hypothyroidism.

Adult

[On the influence of thiocyanates on the delayed immune response in intact and epinephrectomised guinea pigs (author's transl)].

Daily application of 32 mg/kg sodium thiocynate from 1--11th respectively 12th--21st experimental day causes in epinephrectomised guinea pigs at the 26th experimental day an inhibition of cell-mediated immune response (sensitization was induced by intraperitoneal injection of horse serum at the 5th and 7th experimental day). The inhibition of cell-mediated immune response could be demonstrated by means of migration inhibition of spleen cells. Epinephrectomy was performed by operation procedures twice a week. Gluco- and mineralocorticoids were applied supplementary. -- Also other proved thiocyanates respectively compounds with thiocyanate groups such as potassium thiocyanate, ammonium thiocyanate and d,1-2-imino-3.4-dimethyl-5-phenyl-thiacolidinhydrothiocyanate showed the same effect. The results are only pointed out in intact animals, but distinct in epinephrectomised guinea pigs. Finally a significant weaker effect, but of the same kind was observed by potassium- and sodium iodate. Every used dose of prednisolone and cyclophosphamide suppress the migration inhibition. The influence of 2-iminothiacolidin-4-carbon acid and desoxycorticosteronacetat on the migration inhibition is different and depends on the antigen dose applied for sensitization. -- Simulated daily injections undertaken in form of stitches and application of aqua destillata are inefficacious.

Adrenal Glands

Influence of thiocyanate ions on starch-iodine reaction used for the estimation of alpha-amylase activity.

A strong simulating effect of thiocyanate ions during the estimation of alpha-amylase by the amyloclastic method is shown. Thiocyanate ions themselves catalyse the decolorization of the starch-iodine complex. This may lead to wrong values during determination of alpha-amylase by this method. The rate of thiocyanate-iodine reaction depends upon the hydrogen ion concentration of the iodine reagent.

Amylases

Effect of thiocyanate on mammary gland growth in rats.

In growing rats from 3 weeks to 3 months old, 0.1% potassium thiocyanate (KSCN) decreased the plasma protein-bound iodine level by 47% and the total DNA in the mammary gland by 11%, but had no effect on the total RNA. Administration of thiocyanate in the form of 0.1, 0.3 and 0.5% KSCN during pregnancy and 14 d of lactation lowered the protein-bound iodine level by 48-66%, the total DNA by 11-47% and the total RNA by 14-63%, depending on the KSCN dosage. The 14-d-old litters of rats receiving thiocyanate weighed 4-41% less than those in the control group.

Animals

Effect of linamarin on thiocyanate production and thyroid activity in rats.

The principal cyanogenic glucoside in cassava is linamarin (2-hydroxyisobutyronitrile-beta-D-glucoside). Addition of pure linamarin to a diet based on 10% vitamin-free casein did not affect the weight gain of rats. However, as expected, the animals that received the casein diet supplemented with methionine gained more weight than those that received the unsupplemented diet. The amount of thiocyanate ion excreted in the urine by the animals that received linamarin was higher than that excreted by the animals that did not receive linamarin at both levels of dietary methionine. The plasma thiocyanate concentration was also higher for linamarin-fed animals than for animals that did not receive linamarin. Higher plasma thiocyanate levels were associated with a lower percentage uptake of radioiodine by the thyroid.

Animals

A comparison of the effects of sodium thiocyanate and dantrolene sodium on a mammalian isolated skeletal muscle.

1 A combination of electrical and pharmacological stimulation has been used to compare the effects of sodium thiocyanate and dantrolene sodium on the excitation-contraction coupling (ECC) mechanism of the mouse soleus muscle. 2 Thiocyanate prolonged the 'active state' after electrical stimulation and increased the response to 8 mM caffeine and 80 mM KCl. Dantrolene had an opposite effect to thiocyanate on all the indices studied. 3 It is concluded that the mechanism of action of dantrolene is by inhibition of the release of the calcium ions involved in the ECC mechanism, probably at the level of the transverse tubules.

Animals