Role of Amino Acid-catabolizing Enzymes in Urea Synthesis for Rats Treated with the Thyroid Hormone

The purpose of the present study was to determine whether the regulation of urea synthesis was mediated through the supply of nitrogen by amino acid-catabolizing enzymes and whether the concentration of acetylCoA would control the N-acetylglutamate concentration when the thyroid status was manipulated. Experiments were conducted on three groups of rats, each being given 6-propyl-2-thiouracil (PTU, thyroid inhibitor) without a triiodothyronine (T₃) treatment, or PTU + T₃, or neither PTU nor T₃ (control), respectively. The urinary excretion of urea, the liver concentration of N-acetylglutamate, and the hepatic activities of serine dehydratase, threonine dehydratase, alanine transaminase (GPT) and aspartate transaminase (GOT) in rats given PTU + T₃ were significantly lower than those in rats given PTU alone. The activity of glutamate dehydrogenase and the concentrations of free amino acids and acetylCoA in the liver of the PTU + T3-treated group were significantly higher than those in the group treated with PTU alone. These results suggest that the higher activity of amino acid-catabolizing enzymes in the hypothyroid (with PTU alone) rats is likely to stimulate urea synthesis.     

The response of tomato (Lycopersicon esculentum) to different concentrations of inorganic and organic compounds of arsenic

Tomato plants were cultivated in greenhouse and water solutions of arsenite (As(III)), arsenate (As(V)), methylarsonic acid (MA) and dimethylarsinic acid (DMA) were applied individually into cultivation substrate at two As levels, 5 and 15 mg kg⁻¹ of the substrate. Comparing the availability of arsenic compounds increased in order arsenite = arsenate < MA < DMA where the arsenic contents in plants decreased during vegetation period. Within a single plant, the highest arsenic concentration was found in roots followed in decreasing order by leaves, stems, and fruits regardless of arsenic compound applied. Arsenic toxicity symptoms reflected in suppressed growth of plants and a lower number and size of fruits were most significant with DMA treatment. However, the highest accumulation of arsenic by plants growing in the soil containing DMA was caused by higher mobility of this compound in the soil due to its lower sorption affinity. Our results confirmed substantial role of transformation processes of arsenic compounds in soil in uptake and accumulation of arsenic by plants.     

Xenopus laevis tadpole limb regeneration in vivo and in vitro: thyroxine directly promotes blastemal cell proliferation and morphogenesis

Regeneration in hindlimbs of Xenopus laevis larvae which were amputated at stage 53 and 55 through the tarsalia region is promoted by thyroxine (T4), while propyl-thiouracil (PTU) inhibits regeneration when compared to controls. In this paper, by in vivo and in vitro experiments, we demonstrate that the promoting effect of T4 on the regenerative processes of larval X. laevis hindlimbs is a direct effect of this hormone on the blastemal cells. By contrast, the inhibitory effect of PTU on the regenerative process is not due to a direct effect on blastemal cells or to a general toxic effect on the treated larvae, but is related to hypothyroidism induced by the drug. We find that: (i) an increase in blastemal cell proliferation is observed not only in blastemata of T4-treated larvae, but also in blastemata cultured in vitro in a medium supplemented with T4; (ii) the renegerative process is accelerated not only in larvae reared in T4 but also in larvae submitted to a combined treatment of T4 and PTU; (iii) inhibition of cell proliferation is observed in blastemata of PTU-reared larvae but not in blastemata cultured in vitro in a medium supplemented with PTU. Experiments on thyroidless larvae (which were submitted to transplantation of hindlimbs from larvae at stages 53 and 55 followed by amputation of their own right hindlimb and the transplanted limbs) have shown that without thyroid hormone the regenerative process is arrested at cone stage and the promoting effect of T4 treatment is dependent on limb stage and amputation level.     

<Emphasis Type="Italic">Caenorhabditis elegans gcs-1</Emphasis> Confers Resistance to Arsenic-Induced Oxidative Stress

Gamma-glutamylcysteine synthetase (γ-GCS) catalyzes the first, rate-limiting step in the biosynthesis of glutathione (GSH). To evaluate the protective role of cellular GSH against arsenic-induced oxidative stress in Caenorhabditis elegans (C. elegans), we examined the effect of the C. elegans ortholog of GCS(h), gcs-1, in response to inorganic arsenic exposure. We have evaluated the responses of wild-type and gcs-1 mutant nematodes to both inorganic arsenite (As(III)) and arsenate (As(V)) ions and found that gcs-1 mutant nematodes are more sensitive to arsenic toxicity than that of wild-type animals. The amount of metal ion required to kill half of the population of worms falls in the order of wild-type/As(V)>gcs-1/As(V)> wild-type/As(III)>gcs-1/As(III). gcs-1 mutant nematodes also showed an earlier response to the exposure of As(III) and As(V) than that of wild-type animals. Pretreatment with GSH significantly raised the survival rate of gcs-1 mutant worms compared to As(III)- or As(V)-treated worms alone. These results indicate that GCS-1 is essential for the synthesis of intracellular GSH in C. elegans and consequently that the intracellular GSH status plays a critical role in protection of C. elegans from arsenic-induced oxidative stress.     

Modulation of rat liver mitochondrial antioxidant defence system by thyroid hormone

In the present study the effect of thyroid hormone (T(3)) on oxidative stress parameters of mitochondria of rat liver is reported. Hypothyroidism is induced in male adult rats by giving 0.05% propylthiouracil (PTU) in drinking water for 30 days and in order to know the effect of thyroid hormone, PTU-treated rats were injected with 20 microg T(3)/100 g body weight/day for 3 days. The results of the present study indicate that administration of T(3) to hypothyroid (PTU-treated) rats resulted in significant augmentation of oxidative stress parameters such as thiobarbituric acid reactive substances and protein carbonyl content of mitochondria in comparison to its control and euthyroid rats. The hydrogen peroxide content of the mitochondria of liver increased in hypothyroid rats and was brought to a normal level by T(3) treatment. Induction of hypothyroidism by PTU treatment to rats also resulted in the augmentation of total and CN-sensitive superoxide dismutase (SOD) activities of the mitochondria, which was reduced when hypothyroid rats were challenged with T(3). Although CN-resistant SOD activity of the mitochondria remained unaltered in response to hypothyroidism induced by PTU treatment, its activity decreased when hypothyroid rats were injected with T(3). The catalase activity of the mitochondria decreased significantly by PTU treatment and was restored to normal when PTU-treated rats were given T(3). Total, Se-independent and Se-dependent glutathione peroxidase activities of the mitochondria were increased following PTU treatment and reduced when T(3) was administered to PTU-treated rats. The reduced and oxidised glutathione contents of the mitochondria of liver increased significantly in hypothyroid rats and their level was restored to normal when hypothyroid rats were injected with T(3). The results of the present study suggest that the mitochondrial antioxidant defence system is considerably influenced by the thyroid states of the body.     

Changes in thyroid function caused by continuous treatment with thiouracil in rats

Serum concentration evolution of thyroxine and triiodothyronine, and thyrotropin (TSH), have been studied in rats while they were given 6-propylthiouracil (PTU) as antithyroid drug, and during the recovery period after suppression of treatment. In the same way thyroid hypertrophy and plasmatic levels of thyrotropin were correlated. Animals received orally a daily dose of 1 mg/100 g body weight during thirty-five days and had a two week recovery period. Thyroid hormone concentrations in plasma were determined by immunoenzymatic assay ELISA with peroxidase as labelled enzyme. From the results obtained, it can be stated that chronic administration of PTU implies a continuous decrease in thyroid hormone concentrations in plasma, reaching nearly zero values, while after treatment, levels recover their normal values in a week's time. A parallelism exists between thyroid hypertrophy and pituitary TSH hypersecretion, due to a decrease in thyroid hormone levels.     

The <Emphasis Type="Italic">ars</Emphasis> genotype characterization of arsenic-resistant bacteria from arsenic-contaminated gold–silver mines in the Republic of Korea

The isolates were identified on the basis of ars genotype characteristics as well as arsenic oxidation/reduction analysis based on the molecular detection characterization. Diversity, pH range (4.0 to 7.0), location, and ars features were assessed for four arsenic-contaminated pond sites and six arsenic tailings located in the Duck-um mine and Myoung-bong mine areas. The presence of ars genes in the genomes of each bacterial strain was evaluated using polymerase chain reaction. Batch experiment results showed that Pseudomonas putida strains OS-3 and -18 completely oxidized 1 mM of arsenite(III) to arsenate(V) within 35-40 h. In contrast, two arsenate-reducing bacteria isolated from mines, P. putida RS-4 and RS-5, were capable of growing aerobically in growth medium supplemented with up to 66.7 mM arsenate(V), which are significantly higher concentration than those tolerated by other arsenic-resistant bacteria. These results suggest that newly isolated indigenous arsenic-resistant bacteria may provide a better understanding of the molecular geomicrobiology and may be applied to the bioremediation of arsenic-contaminated mines in Korea. Ecologically, the redox potential plays an important role in arsenic toxicity and mobility in As-contaminated mine areas, as it facilitates the biogeochemical cycling activity of Pseudomonas sp. groups.     

Clinical study on early changes in thyroid function of hyperthyroidism treated with propylthiouracil and a relatively small dose of iodide.

In order to compare the acute effects of three kinds of antithyroid agents of iodide (I-), propylthiouracil (PTU) and PTU combined with iodide (PTU+I-) on thyroid function in hyperthyroid patients with diffuse goiter, serum concentrations of thyroxine (T4), triiodothyronine (T3), T3-resin sponge uptake (T3-RU) and free thyroxine index (FT4I) were employed as thyroid function parameters. In the group given iodine (1 mg/day) as iodinated-lecithine, the initial values of T4, T3, T3-RU and FT4I were 20.9 +/- 1.6 microng/100 ml (T4), greater than 740 ng/100 ml (T3), 49.5 +/- 2.3% (T3-RU) and 14.7 +/- 1.8 (FT4I). At the end of one week of therapy, they decreased clearly to 15.6 +/- 2.2 microng/100 ml, 457 +/- 87 ng/100 ml, 42.2 +/- 4.0% and 9.7 +/- 2.4. The so-called "escape phenomenon" from iodide inhibition was observed in serum T4, T3-RU and FT4I values at the end of two weeks of iodide therapy, while serum T3 continued to decrease but the value of T3 was far outside of the normal range. In the PTU group (300 mg/day), thyroid function parameters were 22.5 +/- 0.8 microng/100 ml (T4), greater than 592 ng/100 ml (T3), 54.9 +/- 1.0% (T3-RU) and 18.7 +/- 1.0 (FT4I) before treatment. They decreased continually week by week. At the end of four-week treatment with PTU, the value of each thyroid function parameter was 11.1 +/- 1.9 microng/100 ml, 229 +/- 56 ng/100 ml, 36.6 +/- 4.4% and 5.7 +/- 1.7. In the group of hyperthyroidism simultaneously given both PTU and iodide (300 mg/PTU and 1 mg/iodine), these thyroid function parameters decreased as well as in the group treated with PTU alone for more than two weeks. More rapid or significant decrease of T4, T3, T3-RU and ft4i in PTU+I- group than in PTU group was observed in the present study. These results suggested strongly that iodide alone was not an adequate therapy for hyperthyroidism as well known and they were also compatible with the idea that the concomitant administration of PTU and iodide was more effective in the early phase of therapy of hyperthyroidism than PTU alone.     

Simultaneously Continuous Separation of Glucose,Maltose, and Maltotriose Using a Simulated Moving-Bed Adsorber

The purpose of this study was to find whether the regulation of urea synthesis was mediated through the activation of N-acetylglutamate synthesis by ornithine when the thyroid status was manipulated. Experiments were done on three groups of rats, given 6-propyl-2-thiouracil (PTU, a thyroid inhibitor) without triiodothyronine (T₃) treatment, treated with PTU + T₃, or neither PTU nor T₃ (control). Urinary excretion of urea and the liver concentrations of ornithine and N-acetylglutamate in rats given PTU + T₃ were significantly lower than in rats given PTU alone. The liver concentration of N-acetylglutamate was correlated with the liver concentration of ornithine (r = 0.920, p < 0.001). Ornithine administration (0.5 mmol/100g body wt) elevated the liver concentration of N-acetylglutamate in all three groups. The results suggest that the greater concentration of ornithine in the hypothyroid (PTU alone) rats is likely to increase the N-acetylglutamate concentration in liver and stimulate urea synthesis.     

Protective role of sinapic acid against arsenic: induced toxicity in rats

Arsenic compounds are classified as toxicants and human carcinogens. Environmental exposure to arsenic imposes a big health issue worldwide. Sinapic acid is a phenylpropanoid compound and is found in various herbal materials and high-bran cereals. It has been reported that sinapic acid has antioxidant efficacy as metal chelators due to the orientation of functional groups. However, it has not yet been examined in experimental animals. In light of this fact, the purpose of this study was to characterize the protective role of sinapic acid against arsenic induced toxicity in rats. Rats were orally treated with arsenic alone (5 mg/kg body weight (bw)/day) plus sinapic acid at different doses (10, 20 and 40 mg/kg bw/day) for 30 days. Hepatotoxicity was measured by the increased activities of serum hepatospecific enzymes namely aspartate transaminase, alanine transaminase, alkaline phosphatase, gamma glutamyl transferase, lactate dehydrogenase and total bilirubin along with increased elevation of lipid peroxidative markers, thiobarbituric acid reactive substances, lipid hydroperoxides, protein carbonyl content and conjugated dienes. The toxic effect of arsenic was also indicated by significantly decreased activities of enzymatic antioxidants like superoxide dismutase, catalase, glutathione peroxidase, glutathione-S-transferase, glutathione reductase and glucose-6-phosphate dehydrogenase along with non-enzymatic antioxidant like reduced glutathione. Administration of sinapic acid exhibited significant reversal of arsenic induced toxicity in hepatic tissue. The effect at a dose of 40 mg/kg bw/day was more pronounced than the other two doses (10 and 20 mg/kg bw/day). All these changes were supported by reduction of arsenic concentration and histopathological observations of the liver. These results suggest that sinapic acid has a protective effect over arsenic induced toxicity in rat.     

Characterization of arsenopyrite oxidizing Thiobacillus. Tolerance to arsenite,arsenate, ferrous and ferric iron

Two strains of Thiobacillus, T. ferrooxidans and T. thiooxidans, have been isolated from a bacterial inoculum cultivated during a one-year period in a 1001 continuous laboratory pilot for treatment of an arsenopyrite/pyrite concentrate. The optimum pH for the growth of both strains has been found to be between 1.7 and 2.5. Because of the high metal toxicity in bioleach pulps, the tolerance of T. ferrooxidans and T. thiooxidans with respect to iron and arsenic has been studied. The growth of both strains is inhibited with 10 g/l of ferric ion, 5 g/l of arsenite and 40 g/l of arsenate. 20 g/l of ferrous iron is toxic to T. ferrooxidans but 30 g/l is necessary to impede the growth of T. thiooxidans.     

Ecological restoration of arsenic contaminated soil by Arundo donax L

The possible arsenic tolerance mechanisms were explored in Arundo donax L. under various supplied arsenic concentrations. The treatments included control (no metal) and five doses of arsenic trioxide i.e., 0, 50, 100, 300, 600 and 1000 μg L⁻¹ As to A. donax. The phytoextraction ability of A. donax L. plants was assessed using both the translocation and bioaccumulation factors. The transpirates were collected to analyze the arsenic concentration volatilized along-with study of anatomical characteristics of the plant parts. In general, the arsenite and arsenate accumulation linearly increased in roots, shoot and leaves with the increasing supplied arsenic levels i.e., from 2.348, 2.775 and 3.25 μg g⁻¹ at 50 μg L⁻¹ to 50, 53.125 and 64.25 μg g⁻¹ arsenite, at 1000 μg L⁻¹, from 4.075, 5.425 and 13.56 μg g⁻¹ at 50 μg L⁻¹ to 71, 62.02 and 436.219 μg g⁻¹ arsenate at 1000 μg L⁻¹, respectively. The order of arsenic accumulation in A. donax L. was: solution As(III) < Root As(III) < Shoot As(III) < Leaf As(III) < Solution As(V) < Root As(V) < Shoot As(V) < Leaf As(V). The range of arsenic volatilization by A. donax L. was 7.2–22% at higher supplied arsenic (300–1000 μg L⁻¹). Volatilization was an important mechanism to avoid toxic effects of arsenic by A. donax L. in addition to bioaccumulation.     

Tri-iodothyronine enhances the formation of light mitochondria during cold exposure

The effect of cold exposure and of PTU and PTU + T3 administration on the protein content and succinic dehydrogenase activity of three mitochondrial populations obtained from rat liver was examined. Our results indicated the following: Succinic dehydrogenase activity increases mainly in the light mitochondrial fraction of cold-exposed rats. PTU administration of cold-exposed animals does not affect the increment in enzyme activity of the heavy fraction but blocks the increment of the light fraction. PTU + T3 administration restores succinic dehydrogenase activity to the values prevalent in normal cold-exposed rats. These findings suggest that thyroid hormone may stimulate the formation of light mitochondria during cold exposure.     

Oxidative stress by inorganic arsenic: modulation by thyroid hormones in rat

Arsenic toxicity is attributed mainly to lipid peroxidation and oxidative stress. We therefore studied the modulatory effects of thyroid hormones on arsenic toxicity in rat on lipid peroxidation and oxidative stress. Thyroid hormones, through a mechanism unknown at present, inhibit arsenic accumulation in liver and kidney. Mobilization of arsenic apparently diminishes lipid peroxidation and improves reduced glutathione status, two biochemical demands of combating arsenic toxicity. Results are discussed in reference to the effect of thyroid hormones on microsomal metabolism of arsenic. Arsenic is less toxic in hyperthyroid than in hypothyroid rats. A physiological antagonism between arsenic and thyroxine is discussed.     

Carbohydrate metabolism in growing rice seedlings under arsenic toxicity

We studied in the seedlings of two rice cultivars (Malviya-36 and Pant-12) the effect of increasing levels of arsenic in situ on the content of sugars and the activity of several enzymes of starch and sucrose metabolism: alpha-amylase (EC 3.2.1.1), beta-amylase (EC 3.2.1.2), starch phosphorylase (EC 2.4.1.1), acid invertase (EC 3.2.1.26), sucrose synthase (EC 2.4.1.13) and sucrose phosphate synthase (EC 2.4.1.14). During a growth period of 10-20 d As2O3 at 25 and 50 microM in the growth medium caused an increase in reducing, non-reducing and total soluble sugars. An increased conversion of non-reducing to reducing sugars was observed concomitant with As toxicity. The activities of alpha-amylase, beta-amylase and sucrose phosphate synthase declined, whereas starch phosphorylase, acid invertase and sucrose synthase were found to be elevated. Results indicate that in rice seedlings arsenic toxicity causes perturbations in carbohydrate metabolism leading to the accumulation of soluble sugars by altering enzyme activity. Sucrose synthase possibly plays a positive role in synthesis of sucrose under As-toxicity.     

Effect of propylthiouracil on liver regeneration in rats after partial hepatectomy.

The effect of propylthiouracil (PTU) on the growth activity of intact liver and liver regenerating after partial (65-70%) hepatectomy (PH) was studied in rats. PTU (Propycil, Kali-Chemie, FRG) was dissolved in drinking water (1 g PTU per litre) and this was given to the rats, as their sole source of fluids, three days before PH and then up to the end of the experiment. In rats given PTU, marked inhibition of liver DNA synthesis and the mitotic activity of hepatocytes was found after PH. This effect was potentiated to some extent by partial inanition of the experimental animals given PTU, as demonstrated in a paired feeding test in control rats. PTU inhibition of DNA synthesis in intact and regenerating liver also took effect in thyroidectomized rats, even with substitution (thyroid hormone) therapy. The experiments demonstrated that the effect of propylthiouracil on DNA synthesis in the liver is mediated primarily by way of its direct effect on the liver.     

Seasonal variations of arsenic in mussels<Emphasis Type="Italic"> Mytilus galloprovincialis</Emphasis>

Total arsenic concentration in the edible part of mussels Mytilus galloprovincialis was evaluated seasonally in the coastal area of Rijeka Bay (North Adriatic Sea, Croatia). Sampling stations were located close to the City of Bakar with no industrial facilities (site 1), in the vicinity of the oil refinery and oil thermoelectric power plant (Urinj, site 2), and 4 miles away from the Plomin coal thermoelectric power plant (Brse village, site 3). Additionally, the concentration of arsenic in the tail muscle of the lobster Nephrops norvegicus, collected in Rijeka Bay, was studied. During winter at sites 2 and 3, the total arsenic in the edible part of the mussels was 16.4 mg As/kg FW (FW=fresh weight) and 4.38 mg As/kg FW, respectively, and increased during springtime at site 2 (6.5 mg As/kg FW) compared to the rest of the year, when individual total arsenic concentration at all sites ranged from 1.7 to 3.7 mg As/kg FW. In the winter (sites 2 and 3) and springtime (site 2) there was no correlation between the length of the mussel shell and the arsenic concentration in the edible part of the mussels. In the other seasons, at sites 1, 2 and 3, there was a correlation between arsenic in the edible part of mussels and shell length in most cases (correlation coefficients r varied from 0.64 to 0.85; P <0.05 to P <0.01). Correlation between shell length (in the narrow range of shell lengths from 3.4 to 5.0 cm) and arsenic in the edible part of the mussels shows linearity with a high regression coefficient ( r =0.914; P <0.001). The increase of arsenic in the mussels during winter and spring was suggested at least partially as a result of a low nutritional status, i.e. reduced weight of the mussels edible part during winter. In addition, a linear relationship was found between body length and arsenic concentration in the tail muscle (mean 17.11±4.48 mg As/kg FW) of the Norway lobster.Communicated by H.-D. Franke     

Relationship of biochemical and morphological changes in rat thyroid and proton spin-relaxation of the tissue water.

The effect was studied of biochemical and morphological changes induced by antithyroid substances (PTU, C10(-4)) on proton spin-relaxation properties of rat thyroid gland. It was found that thyroid stimulated by PTU (0.05%) or C10(-4) (1.0%) exhibit marked morphological changes (hyperplasia and epithelial hypertrophy) with alteration of the soluble iodoprotein pattern (content and composition.). Both relaxation times spin-lattice (T1) and spin-spin (T2) were increasing with the lenght of treatment with antithyroid drugs. Reversibility of the process was noted in accordance with biochemical and morphological data. The relaxation rate (formula: see text) for thyroid tissue water was in positive correlation with the suluble protein concentration and particularly with the TG content in the gland. There was no difference in relaxation times between normal thyroid and gland of rats treated chronically with excess iodide. The observed difference in T1 between normal glands and glands of PTU,-C10(-4)--treated rats was comparable with that found in cases of human thyroid cancer. This finding is of importance when the diagnostic potential of NMR in the detection of malignancy is considered. In conclusion, a strong correlation was found between microstructural and biochemical changes of the thyroid gland and proton magnetic relaxation of tissue water. The striking difference between the proton spin-relaxation times in normal and in goiter thyroid glands of rats suggests that pulsed NMR spectroscopy could be a method for evaluation of some disturbances in thyroid gland.     

Sequestering of As(III) and As(V) from wastewater using a novel neem leaves/MnFe2O4 composite biosorbent

An arsenic biosorbent comprising neem leaves (NL) and MnFe₂O₄ particles was developed and its removal potential was investigated. Physicochemical analysis of the NL/MnFe₂O₄ composite (MNL) was performed for the Brunauer, Emmett and Teller surface area, Fourier transform infrared spectra (FT-IR), and scanning electron microscopy–Energy-dispersive X-ray (EDX). The following parameters were optimized: pH, biosorbent dose, contact time, temperature, and initial arsenic concentration. The optimum pH values achieved for biosorption of As(III) and As(V) were 7.0 and 4.0, respectively, when the equilibrium time was 110 minutes for both. MNL was found to be efficient with 85.217% and 88.154% biosorption efficiency at a concentration of 50 mg/L of As(III) or As(V) solution, respectively. This was also proved by the FT-IR study of arsenic-loaded biosorbent. For establishing the best suitable correlation for the equilibrium curves exploiting the procedure of the nonlinear regression for curve fitting analysis, isotherm studies were conducted for As(III) and As(V) using 30 isotherm models. The pattern of biosorption fitted well with Brouers–Sotolongo isotherm model for As(III) and Langmuir–Freundlich as well as Sips isotherm models for As(V). Dubinin–Radushkevich (D-R) isotherm studies specified that ion exchange might play a significant role. The influence of various co-existing ions at different concentrations was examined. Desorption study was performed using various concentrations of NaOH solution.