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Effects of red cell exchange on calculated sheep lung vascular permeability to 14C-urea and 14C-thiourea.

We performed fourteen experiments on sheep to determine whether or not erythrocytes tracer transport alters the calculated value of capillary tracer permeability surface area. In each set of experiments 14C-urea or 14C-thiourea was equilibrated with: (1) a whole blood sample; (2) saline alone, and (3) packed erythrocytes alone. Aliquots of each of these samples were injected separately into the superior vena cava of sheep and multiple indicator data collected from the aorta. Lung urea or thiourea microvascular permeability surface area (PS) was calculated for each set of data using the integral extraction method. These results were compared to the predictions of a detailed theory of microvascular transport which included red cell effects. As predicted by the theory, only small differences were found between urea PS calculations based on pre-equilibrated blood or plasma injectates. When 14C-urea in the injectate was confined to red cells, the average PS calculation was approximately 60% of the average plasma-equilibrated value (also predicted by theory). This ratio was substantially lower (25%) when 14C-thiourea was used as an indicator, suggesting that the red cell membrane, rather than the microvascular barrier, limits thiourea exchange. We conclude that a finite red cel-plasma urea exchange rate does not significantly influence the calculation of lung vascular PS when the injected blood is equilibrated with urea prior to its introduction into the vascular system.

Animals↗

Covalent binding of [14C]thiourea to protein in lungs from endotoxin-treated rats.

Administration of thiourea to mature male rats at a dosage of 3.5 mg/kg (ip) produced marked pleural effusion by 2 hr (3-4 ml). Pretreatment with bacterial lipopolysaccharide (endotoxin) significantly reduced this pleural effusion (less than 0.5 ml). Despite this unequivocal effect, there was no corresponding reduction in the covalent binding of [14C]thiourea to lung protein. These data indicate that the protective effect of endotoxin against the initial stages of thiourea pneumotoxicity does not involve a reduction in binding of the [14C]thiourea or a metabolite to lung protein. However, alterations in low levels of binding to specific cell types or particular protein(s) relative to covalent binding cannot be ruled out.

Animals↗

High-pressure liquid chromatography separation of phenobarbitone, thiourea, and thioacetamide of toxicological interest.

It was possible, by reverse-phase high-pressure liquid chromatography (HPLC) to separate thiourea, thioacetamide, and phenobarbitone with retention times of 1.4, 1.7, and 4.7 min, respectively. Thiourea and thioacetamide are known carcinogens. Substances such as phenobarbitone, thiourea, and thioacetamide, which may be encountered in our environment, can now be identified by this technique. It is also recommended that this HPLC technique should be used routinely in the screening of the purity of substances which are being subjected to toxicity, carcinogenicity, mutagenicity, and teratogenicity evaluation. A mixture of impurities in the sodium phenobarbitone was removed in the synthesis of the 5-ethyl-5-phenyl-barbituric acid (phenobarbitone). These substances which were not thiourea or thioacetamide could have been responsible for the positive result in the in vitro degranulation test for carcinogens with sodium phenobarbitone.

Abnormalities, Drug-Induced↗

Regeneration experiments of the platinated enzyme fumarase, using sodium diethyldithiocarbamate, thiourea, and sodium thiosulfate.

The enzyme fumarase is inhibited by [cis-Pt(NH3)2(H2O)2] (NO3)2. The Pt compound most likely binds at a S-methionine site. Sodium diethyldithiocarbamate (Naddtc) appears to be a powerful regenerator of enzymatic activity. Thiourea is less active, while sodium thiosulfate (STS) is almost inactive in restoring the activity of the enzyme. The regeneration phenomena are based on the dissociation of the Pt-S bonds of the methionine type, and formation of species like [Pt(ddtc)2]. In the model adduct [Pt(dien)GS-Me]2+ Naddtc, thiourea and STS easily break the Pt-S bond of the methionine type. It is concluded that the model system for Naddtc and thiourea does resemble fumarase quite well. S-donor ligands, which may be used as rescue agents in Pt antitumor therapy, are known to suppress nephrotoxicity caused by [cis-PtCl2(NH3)2]. A parallel is drawn between the enzyme reactivation, modeled by fumarase, and the [cis-PtCl2(NH3)2] nephrotoxicity suppression by rescue agents. It is proposed that a Pt-methionine type binding is broken by the rescue agents Naddtc and thiourea, but that the rescue agent STS only inhibits the nephrotoxicity by inactivating unbound Pt species in the cell.

Animals↗

Assessment of contact sensitivity of four thiourea rubber accelerators: comparison of two mouse lymph node assays with the guinea pig maximization test.

Contact sensitivity of four thiourea rubber accelerators, diphenylthiourea (DPTU), dilaurylthiourea (DLTU), dibutylthiourea (DBTU) and diethylthiourea (DETU), was evaluated by a new sensitive mouse lymph node assay (SLNA) and the murine local lymph node assay (LLNA). The results of the SLNA and LLNA were compared with the data of the guinea pig maximization test (GPMT). In the LLNA and SLNA, the sensitizing activity was measured as a function of draining lymph node activation following application of the test chemicals. Of these four thioureas, three (DETU, DBTU and DPTU) were not classified as skin sensitizers in the LLNA. The SLNA successfully detected the sensitivity of all thioureas tested. This result indicated that the SLNA was, in these cases, more sensitive than the LLNA for identification of contact allergens. The order of sensitization potential observed from the SLNA was DPTU (greatest), DLTU, DBTU and then DETU (least). The predictions of sensitizing potential and the order of the sensitizing capacity of four thioureas by the SLNA and the GPMT are very similar.

Administration, Topical↗

Analysis of structure-activity relationships for the 'B-region' of N-(3-acyloxy-2-benzylpropyl)-N(')-[4-(methylsulfonylamino)benzyl]thiourea analogues as vanilloid receptor antagonists: discovery of an N-hydroxythiourea analogue with potent analgesic activity.

The structural modifications on the B-region of the potent and high affinity vanilloid receptor (VR1) lead ligand N-(3-acyloxy-2-benzylpropyl)-N(')-[4-(methylsulfonylamino)benzyl]thiourea were investigated by the replacement of the thiourea with diverse isosteric functional groups. Structure-activity analysis indicated that the A-region in this series was the primary factor in determining the agonistic/antagonistic activities regardless of the B-region. The N(C)-hydroxy thiourea analogues (12, 13) showed excellent analgesic activities in the acetic acid writhing assay compared to the parent thiourea analogues.

Analgesics↗

Thiourea-induced thyroid hormone depletion impairs testicular recrudescence in the air-breathing catfish, Clarias gariepinus.

We used thiourea-induced thyroid hormone depletion as a strategy to understand the influence of thyroid hormones on testicular recrudescence of the air-breathing catfish, Clarias gariepinus. Treatment with 0.03% thiourea via immersion for 21 days induced hypothyroidism (thyroid hormone depletion) as evidenced by significantly reduced serum T(3) levels. Thiourea-treated males had narrowed seminiferous lobules with fewer spermatozoa in testis, very little or no secretory fluid, reduced protein and sialic acid levels in seminal vesicles when compared to controls. The histological changes were accompanied by reduction in serum and tissue levels of testosterone (T) and 11-ketotestosterone (11-KT), a potent male specific androgen in fish. Qualitative changes in the localization of catfish gonadotropin-releasing hormone (cfGnRH) and luteinizing hormone (LH, heterologous system) revealed a reduction in the distribution of immunoreactive neuronal cells and fibers in thyroid depleted fish. Interestingly, thiourea-withdrawal group showed physiological and histological signs of recovery after 21 days such as reappearance of spermatozoa and partial restoration of 11-KT and T levels. These data demonstrate that thyroid hormones play a significant role in testicular function of catfish. The mechanism of action includes modulating sex steroids either directly or through the hypothalamo (GnRH)-hypophyseal (LH) axis.

Animals↗

Thin-layer polarographic detector for the high-performance liquid chromatographic detection of thiourea derivatives.

A polarographic thin-layer detector for high-performance liquid chromatography, was devised. The detector has a sandwich structure with a working surface area of the mercury electrode of 0.25 mm2. The detector was applied to the direct amperometric detection and for detection with reactivation of the electrode surface by voltage pulses. It is shown that the latter detection mode is suitable for the determination of compounds that form insoluble or complex mercury salts, e.g., thiourea. Thiourea and its derivatives can be separated and determined in a wide concentration range and the detection limit for thiourea corresponds to 2 ng injected. Thiourea can be determined directly in urine.

Chromatography, High Pressure Liquid↗

Free radicals generated in yeast by the Salmonella test-negative carcinogens benzene, urethane, thiourea and auramine O.

A large fraction of carcinogens score negative in short-term genotoxicity assays such as the Salmonella reverse mutation (Ames) assay. More information is needed about the mechanism of action of such Salmonella-negative carcinogens. Many Salmonella-negative carcinogens induce deletions due to intrachromosomal recombination in Saccharomyces cerevisiae with an apparent threshold. We have previously shown that the Salmonella-negative carcinogens cadmium, aniline, chloroform and carbon tetrachloride generate free radical species in S. cerevisiae. We have further investigated the possible generation of intracellular free radical species by the diverse Salmonella-negative carcinogens benzene, urethane, thiourea and auramine O. The toxicity and recombinagenicity of thiourea and auramine O was reduced in the presence of the free radical scavenger N-acetyl cysteine. N-acetyl cysteine did not protect against toxicity or recombination induced by the Salmonella-positive carcinogens ethyl methane sulfonate, methyl methane sulfonate or nitroquinoline-N-oxide. A strain deficient in the enzyme superoxide dismutase, which catalyses the dismutation of superoxide anion radical, was hypersensitive to killing by benzene, urethane and thiourea. The sod- strain was only slightly more sensitive to the Salmonella-positive carcinogens. Intracellular oxidation of the free radical-sensitive reporter compound dichlorofluorescin diacetate was increased in yeast cultures exposed to benzene, urethane and auramine O; again, the Salmonella mutagens had no effect on oxidation of the dye. These data show that free radical species are produced in Saccharomyces cerevisiae following exposure to benzene, urethane, thiourea and auramine O, and suggest a possible role for oxidative stress is recombination induced by these carcinogens.

Acetylcysteine↗

Nonpeptide somatostatin agonists with sst4 selectivity: synthesis and structure-activity relationships of thioureas.

Utilizing NNC 26-9100 (11) as a structural lead, a variety of nonpeptide derivatives of somatostatin were synthesized and evaluated for sst2 and sst4 receptor binding affinity. A novel thiourea scaffold was utilized to attach (1) a heteroaromatic nucleus to mimic the Trp8 residue, (2) a nonheteroaromatic nucleus to mimic Phe7, and (3) a primary amine or other basic group to mimic the Lys9 residue of somatostatin. Displacement studies were carried out using membranes from cell lines expressing ssts [BHK cells (sst4) and HEK 293 cells (sst2)] utilizing [125I]Tyr11-SRIF as the radioligand. Several thioureas (11, 38, 39, 41, and 42) and the urea 66 exhibited Ki values of less than 100 nM. The thioureas 11 (Ki = 6 nM) and 41 (Ki = 16 nM) and the urea 66 (Ki = 14 nM) are believed to be the most potent nonpeptide sst4 agonists known. Since the thiourea 11 and the urea 66 exhibit high sst4 selectivity, these novel nonpeptide derivatives may be useful tools for studying the sst4 receptor. Studies are currently in progress to evaluate the therapeutic potential of NNC 26-9100 (11) in the treatment of glaucoma.

Aminopyridines↗

Plan-view and cross-sectional characterization of thiourea-treated phosphorus-added steel surface.

The galvanizing reaction is retarded when phosphorus-added steel is used as a substrate. We have found that both the galvanizing and the galvannealing reactions are highly promoted when thiourea solution is coated on the cold-rolled steel surface before the annealing process. Both a plan-view and cross-sectional characterization of thiourea-coated steel was performed using low-voltage scanning electron microscopy and a focused ion beam. A fine grain structure is formed in the surface region of approximately 1 microm in thickness. In this region, (Mn, Fe)S particles are formed by the reaction between sulphur from thiourea and manganese from steel. These (Mn, Fe)S particles have a pinning effect on the cold-rolled steel grains and greatly retard recrystallization when the steel is annealed. It is concluded that the promotion of the galvannealing reaction is due to the increased diffusion paths of zinc and iron materialized by the fine grain structure of the thiourea-coated steel.

Microscopy, Electron, Scanning↗

Structural requirements for potent anti-human immunodeficiency virus (HIV) and sperm-immobilizing activities of cyclohexenyl thiourea and urea non-nucleoside inhibitors of HIV-1 reverse transcriptase.

The current pandemic of sexually transmitted human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) has created an urgent need for a new type of microbicide, one that is both a spermicide and a virucide. In a systematic effort to identify a non-detergent-type antiviral spermicide, we have rationally designed and synthesized a series of cyclohexenyl thiourea (CHET) nonnucleoside inhibitors (NNIs) of HIV-1 reverse transcriptase (RT) with sperm-immobilizing activity (SIA). To gain further insight into the structural requirements for the optimal activity of these dual-function NNIs, we compared the effects of thiazolyl, benzothiazolyl, and pyridyl ring substitutions and functionalization with electron-donating and electron-withdrawing groups as well as the importance of thiourea and urea moieties of 15 heterocyclic ring-substituted NNIs. RT activity and p24 antigen production in HIV-infected peripheral blood mononuclear cells were used as markers of viral replication. Computer-assisted sperm analysis was used for evaluating SIA of CHET compounds. The rabbit model was used for evaluation of in vivo mucosal toxicity and contraceptive activity of the lead NNIs. Three CHET-NNIs with a bromo, chloro, or methyl substitution at the 5 position of the pyridyl ring exhibited potent anti-HIV activity at nanomolar concentrations (IC(50) = 3-5 nM) and SIA at micromolar concentrations (EC(50) = 45-96 micro M). The dual-function CHET-NNIs were potent inhibitors of drug-resistant HIV-1 strains with genotypic and phenotypic NNI resistance. Upon substitution of the sulfur atom of the thiourea moiety with an oxygen atom, the most striking difference noted was a 38-fold reduction in time required for 50% sperm immobilization (T(1/2)). A quantitative structure-activity relationship (QSAR) analysis was used in deriving regression equations between 20 physicochemical properties and SIA of NNIs. QSAR analysis showed that the T(1/2) values positively correlated with values for molecular refractivity (r = 0.88), hydrophobicity (r = 0.72), atomic polarizability (r = 0.70), and principal moment of inertia (r = 0.63) of spermicidal NNIs. A stepwise multiple regression model to describe the relationship of T(1/2) values with these four regressors provided excellent predictability (r = 0.93). Exposure of semen to thiourea/urea NNIs either alone or in combination at the time of artificial insemination led to marked or complete inhibition of pregnancy in rabbits as assessed by the number of embryo implants versus corpora lutea on Day 8 of pregnancy. Repeated intravaginal application of a gel-microemulsion with and without 0.5%, 1%, and 2% CHET-NNI or its urea analog either alone or in combination did not induce mucosal toxicity. We hypothesize that the gain of spermicidal function by CHET-NNIs is due to their metabolic oxidation to urea analogs by sperm. Three reaction pathways are discussed. The extremely rapid SIA of the urea analog as well as the broad-spectrum anti-HIV activity of spermicidal CHET-NNIs together with their lack of mucosal toxicity and the marked ability to reduce in vivo fertility is particularly useful for the clinical development of a dual-function spermicidal microbicide. The cyclohexenyl pyridyl NNIs, especially N-[2-(1-cyclohexenyl)ethyl] N'-[2-(5-bromopyridyl)]-thiourea in combination with the urea analog, show unique clinical potential as anti-HIV spermicides aimed at curbing the sexual transmission of multidrug-resistant HIV-1 while providing effective fertility control for women.

Aminopyridines↗

The change of disposition kinetics for thiourea accompanied by growth in mice.

The whole blood levels and whole-body autoradiography following intravenous administration of thiourea, which is considered to covalently bind to the macromolecules in the body, were investigated in 1-day-old, 1-week-old, 3-week-old and 8-week-old mice in order to clarify the change of disposition kinetics accompanied by growth. The total body clearance for thiourea was remarkably lower in 1-day-old and 1-week-old mice than in more aged mice. The lower metabolic clearance as well as renal clearance in 1-day-old and 1-week-old mice seemed to be responsible for this observation. The whole-body autoradiograms obtained at 60 min following intravenous administration of 14C-thiourea showed the homogeneous distribution of radioactivity throughout the body in 1-day-old and 1-week-old mice and the high radioactivity localized in the liver and lung in more aged mice. This remarkable difference was assumed to be brought about by the difference in quantity of the macromolecules in the liver and lung which can covalently bind with thiourea. This assumption was supported by the result of gel filtration on Sephadex G-10 of the liver and lung homogenates.

Aging↗

Effect of hypophysectomy by partial decapitation and treatment with thiourea on iodine metabolism in the developing chick embryo.

The hypothesis of an action of the pituitary gland of the developing chick embryo in the transfer of iodide from the yolk of the egg to the circulation of the embryo, through the yolk sac, was tested. Plasma iodide levels and thyroidal iodine contents were determined in hypophysectomized (by partial decapitation), thiourea-injected and control embryos. From day 11 of incubation these parameters were always lower in the 'hypophysectomized' embryos than in controls, and plasma iodide levels of the thiourea-treated embryos were higher than those of controls. These results indicate a reduced iodide transfer from the yolk to the 'hypophysectomized' embryo, and an increased iodide transfer to the thiourea-treated embryo. This occurred in spite of a reduced thyroid hormonal secretion in both series. The pituitary gland could therefore have a direct action (not through the thyroid gland) at the yolk sac level, to augment the transfer of iodide from the yolk in intact embryos from day 11 to the end of incubation. Thyroid-stimulating hormone (TSH) could be the pituitary hormone acting at the yolk sac level, the increased iodide transfer observed in the thiourea-injected embryos being due to a raised TSH secretion responding to the decreased plasma thyroxine levels.

Animals↗

Effects of dimethyl sulfoxide and thiourea upon intercalator-induced DNA single-strand breaks in mouse leukemia (L1210) cells.

The free radical scavengers, dimethyl sulfoxide (Me2SO) and thiourea, were used to assess the role of free radicals in the production of intercalator-induced DNA breaks and cytotoxicity in mouse leukemia L1210 cells. Both agents decreased X-ray break production, and this decrease was comparable in magnitude to the degree of inhibition of X-ray-induced cell killing. By contrast, Me2SO increased the DNA breaks produced by the intercalators, Adriamycin, 5-iminodaunorubicin, and 4'-(9-acridinylamino)methanesulfon-m-anisidide. This was not due to an enhancement of Adriamycin or 4'-(9-acridinylamino)methanesulfon-m-anisidide uptake by Me2SO. Strand break production by intercalators was decreased by thiourea. This was not due to an inactivation of the intercalators or to a decrease of Adriamycin or 4'-(9-acridinylamino)methanesulfon-m-anisidide uptake by thiourea. Experiments using nucleoid sedimentation to assess the DNA linking number and domain size from cells treated with Me2SO and thiourea indicated that these chemicals alter chromatin structure in a fashion which may account for effects on intercalator-induced DNA scission. The alterations in intercalator-induced DNA scission were not accompanied by corresponding alterations in cytotoxicity, thus dissociating intercalator-induced strand break production from lethality and the mechanism of X-ray break production.

Animals↗

[Structurally dependent effects of substituted thioureas on rna phage m 12 (author's transl)].

Using the agar diffusion method, thiourea as well as N-aryl-, N, N'-diaryl-, N-aryl-N'-alkyl-, N,N'-dialkyl-, and N-alkyl-substituted thiourea derivatives were examined for their effects on the system of Escherichia coli W 1665 F+/M 12. The compounds tested were found to produce more or less strongly inhibitive or promotive effects on the RNA phages and/or host bacteria. N-phenyl-N'-dibutyl thiourea, N-phenyl-N'-acetyl thiuorea, and N1,N1'-p-phenylene-bis-(N3-n-propyl thiourea) showed an intense inhibitory effect on M12. Quite obviously, the decisive intervention by these substances in the multiplication of viruses was by a replicative step subsequent to the adsorption of phages.

Coliphages↗

Synthesis of a series of new N1-[4-(4-nitrophenylthio)phenyl]-N3-(H/alkyl/acyl/aryl) thioureas and their antifungal, insecticidal and larvicidal activities.

A series of new mono and disubstituted thioureas, derived from 4-(4-nitrophenylthio) aniline, have been prepared and screened for antifungal activity against Alternaria alternata and Curvularia lunata. Their insecticidal and larvicidal activities have also been evaluated against adult male and female cockroaches (Periplanata americana) and mosquito larvae (Culex pipiens fatigans Wied) of IIIrd and early IVth instar stage, respectively. The tabulated results reveal that most of the thiourea derivatives, in general, possess marked antifungal, insecticidal and larvicidal properties, but the chlorosubstituted derivatives, in particular, are the most active compounds in the entire series. Compound 20, N1-[4-(4-nitrophenylthio)phenyl]-N3-(4-chlorophenyl) thiourea, was the most active antifungal compound, whereas, compound 12, N1-[4-(4-nitrophenylthio)phenyl]-N3-(4-chlorobenzoyl) thiourea, was the most potent insecticidal and larvicidal compound.

Alternaria↗

Development of N-benzamidothioureas as a new generation of thiourea-based receptors for anion recognition and sensing.

A series of neutral N-(substituted-benzamido)-N'-phenylthioureas (substituent = p-OC(2)H(5), p-CH(3), m-CH(3), H, p-Cl, p-Br, m-Cl, and p-NO(2)) were designed as anion receptors, in which the thiourea binding site was attached to the benzamido moiety via an N-N bond. The absorption spectra of these N-benzamidothioureas in acetonitrile peaked at ca. 270 nm were found to show unprecedented red shifts by 7 373 to 14 325 cm(-1) in the presence of anions such as AcO(-), F(-), and H(2)PO(4)(-). Under the same conditions, the classic neutral thiourea receptors, N-(substituted-phenyl)-N'-phenylthioureas, showed absorption spectral shifts in most cases of less than 800 cm(-1) with one exception of 6501 cm(-1). Control experiments, effects of protic solvent, and (1)H NMR titration confirmed the formation of hydrogen-bonding complexes between the new N-benzamidothiourea receptors and anions. The binding constants with AcO(-), for example, are at 10(5)-10(7) mol(-1) L order of magnitude, which are 13 to 590 times those of the corresponding classic N-phenylthioureas in the same solvent. It was found that, whereas the absorption of the N-benzamidothiourea receptors showed essentially no dependence on the substituent, the substantially red-shifted new absorption band of the N-benzamidothiourea-anion binding complex was sensitively subject to the substituent. A linear relationship was found between the absorption energies of the N-benzamidothiourea-acetate binding complexes and the Hammett constants of the substituents with a negative slope of -0.34 eV. This led to the assignment that the substantially red-shifted absorption band was the ground-state intramolecular charge-transfer absorption with the substituent locating in the electron acceptor moiety. It was concluded that anion binding to the thiourea moiety of the N-benzamidothiourea receptors switched on their ground-state charge transfer. An anion-binding induced structural change was suggested to occur around the N-N bond in N-benzamidothioureas, which resulted in a substantially increased electron donating ability of the electron donor in the receptor molecules. As a consequence, the ground-state charge transfer takes place in the N-benzamidothiourea-anion binding complexes, leading to unprecedented red shifts in the absorption spectra and substantially enhanced anion binding affinities than those of the corresponding N-phenylthiourea receptors. N-Benzamido-N'-phenylthioureas represent a new generation of neutral thiourea-based anion receptors that show substantially improved anion binding performance important for anion sensing and recognition.

Journal Article↗