Thyroid peroxidase as toxicity target for dithiocarbamates

In vivo ethylenebisdithiocarbamates and ETU are toxic to the thyroid gland. Since the molecular target of these compounds is thought to be thyroid peroxidase (TPO) which catalyzes the transfer of iodine to thyroglobulin, we examined the effect of these compounds on peroxidative activity in Chinese hamster ovary (CHO) cells transfected with the human TPO gene. The activity was inhibited by 50 microM ETU, 5 microM ziram and 5 microM zineb, the last-mentioned effect being irreversible in the absence of iodide. Thiram had no effect. By contrast, the iodinating activity of TPO was blocked only by 5 microM ETU and 50 microM zineb but not by the other compounds. The effect on TPO-catalysed iodination could explain the differences in thyrotoxicity of these compounds in vivo.     

Thyroid function in rats with iodine deficiency is not further impaired by concurrent, marginal zinc deficiency

The hypothesis tested was that Zn deficiency aggravates impaired thyroid function as induced by I deficiency. In two separate experiments male rats were fed on diets either deficient in Zn or in I, or deficient in both. An identical, restricted amount of food was given to each rat so that body-weight gain of the experimental groups was comparable. Zn deficiency was evidenced by reduced tibial Zn concentrations. I deficiency was evidenced by goitre, reduced urinary I excretion, reduced plasma thyroxine concentrations and reduced absolute amounts and concentrations of thyroxine in the thyroid. Zn deficiency had no effect on the raised thyroid weight as induced by I deficiency. Zn restriction from 184 mumol Zn/kg diet to 31 mumol Zn/kg diet, but not to 92 mumol Zn/kg diet, significantly lowered plasma thyroxine concentration. There were no interrelated effects of Zn and I deficiencies on thyroid hormone levels. These results indicate that marginal Zn deficiency does not influence thyroid hormone metabolism in I deficiency.     

Plasma somatomedin activity, growth-hormone and insulin levels in vitamin B6 deficient rats

Plasma somatomedin activity was low in rats fed a vitamin B6 deficient diet for 4 weeks. The reduction appears to be in the fraction which is not growth hormone dependent. Plasma growth hormone levels were unaltered while plasma insulin levels were significantly reduced.     

Thyroid function in a population with an extra iodine intake]

Polymorphonuclear leukocyte (PMN) superoxide (.O2-) production has been implicated in the pathogenesis of cardiopulmonary bypass (CPB)-related end organ injury. PMN "priming" has been described as an event which enhances the release of .O2- following a second, activating insult. We hypothesized that PMN priming occurs during CBP and is temporally related to the plasma level of complement (C3a), interleukin (IL)-6, and IL-8. PMNs were isolated from 10 CPB patients pre-bypass (preCPB), 5 min after protamine administration (PROT), and at 6 and 24 h post-CPB. PMN .O2- production was measured by a cytochrome c reduction assay in the presence or absence of either phorbol 12-myristate-13-acetate (PMA, 0.4 microgram/ml) or N-formyl-methionyl-leucyl-phenylalanine (FMLP, 1 microM) and also after priming with 2000 nM platelet-activating factor (PAF) followed by activation with either PMA or FMLP. Plasma levels of C3a, IL-6, and IL-8 were determined by enzyme-linked immunosorbent assay. PMA-activated PMN .O2- production was significantly elevated at 6 h post-CPB compared to pre-CPB levels (11.04 +/- 0.9 vs 7.62 +/- 0.57, P = 0.009), indicating that CPB is associated with in vivo PMN priming. When PMNs were primed in vitro with PAF and then activated with PMA or FMLP, .O2- release at 6 h post-CPB was also significantly greater than pre-CPB levels (16.04 +/- 0.74 vs 12.2 +/- 0.92, P = 0.038; and 17.33 +/- 1.38 vs 13.33 +/- 1.35, P < 0.05), indicating that CPB acts synergistically with PAF to prime PMNs. Levels of C3a rose significantly over pre-CPB levels at PROT (P = 0.001), and IL-6 and IL-8 rose over pre-CPB levels at 6 h post-CPB (P = 0.01 and P = 0.006, respectively). These findings demonstrate that CPB not only directly primes PMNs, but also potentiates priming of PMNs by PAF. This "primed" PMN state, which coincided with the increased plasma levels of inflammatory mediators, may suggest a mechanism of predisposition to organ dysfunction following CPB.     

Thyroid peroxidase: kinetics, pH optima and substrate dependency

The oxidation of iodide, guaiacol and 2,2'-azino-di[3-ethyl-benzthiazoline-(6)-sulphonic acid] and the iodination of tyrosyl residues in bovine serum albumin, catalysed by partly purified thyroid peroxidase, were studied. The enzyme showed pH optima with all electron donors. With the exception of guaiacol, the position of the pH optima depended upon both the electron donor and hydrogen peroxide concentrations. With increased hydrogen peroxide concentrations the optima shifted to lower pH, and with increased iodide concentration to higher pH. For monoiodotyrosine (MIT) formation in bovine serum albumin the position of the pH optimum was also dependent on the hydrogen peroxide concentrations. The position of the pH optimum of the oxidation of guaiacol was pH 9 and independent of substrate and hydrogen peroxide concentrations. It is obvious from these findings that iodination reactions must be studied under well-defined conditions.     

Fetal thymus development in vitamin D deficient rats

A specifically designed handpiece has been developed for ultrasonic dissection of tissues and organs during minimal-access surgery. The experimental prototype has been evaluated in major endoscopic operations on the esophagus, colon, and rectum (n = 19). The benefits documented by this initial experience include increased dissection efficiency of extensive fibroareolar attachments, safe exposure of major vascular pedicles (especially those located in mesocolic fat), greatly reduced risk of major hemorrhage, and decreased operating time.     

Linoleic acid desaturation activity of liver microsomes of essential fatty acid deficient and sufficient rats

By employing fluorescent histochemistry and spectrofluorimetry the influence of an original psychotropic substance -- carbidine on the level, localization and accumulation of the adrenergic mediator in Vas deferens, dura mater and the kidney capsule was studied "in vivo" and "in vitro". Carbidine was shown to capable of liberating the adrenergic mediator from the sympathetic nerve fibres of the peripheral tissues without affecting the capture and accumulation of exogenous norepinephrine by adrenergic nerve fibres.     

Thiol-linked peroxidase activity of human ceruloplasmin

Human ceruloplasmin exhibited different antioxidant effects according to the electron donors in a metal-catalyzed oxidation system. Purified ceruloplasmin did not play a significant role in the protection of DNA strand breaks in the ascorbate/Fe3+/O2 system. However, when ascorbates were replaced with a thiol-reducing equivalent such as dithiothreitol, DNA strand breaks were significantly prevented by the same amount of ceruloplasmin. Ceruloplasmin did not catalyze the decomposition of H2O2 in the absence of reduced glutathione. On the contrary, ceruloplasmin showed a potent peroxidase ability to destroy H2O2 in the presence of reduced glutathione. In conclusion, the removal of H2O2 by human ceruloplasmin is not simply stoichiometric but thiol-dependent.     

Localization of peroxidase activity in Trypanosoma cruzi microbodies

Electron microscopic observation of Trypanosoma cruzi epimastigotes reveals the presence of microbody-like structures (microperoxisomes) in which 3,3'-diaminobenzidine (DAB) is peroxidized to electron-opaque material. The role of peroxidase in DAB peroxidation is supported by the enzyme demonstration in disrupted epimastigotes and the microbody-containing cell fractions.     

Mice deficient in cellular glutathione peroxidase develop normally and show no increased sensitivity to hyperoxia

Glutathione peroxidase, a selenium-containing enzyme, is believed to protect cells from the toxicity of hydroperoxides. The physiological role of this enzyme has previously been implicated mainly using animals fed with a selenium-deficient diet. Although selenium deficiency also affects the activity of several other cellular selenium-containing enzymes, a dramatic decrease of glutathione peroxidase activity has been postulated to play a role in the pathogenesis of a number of diseases, particularly those whose progression is associated with an overproduction of reactive oxygen species, found in selenium-deficient animals. To further clarify the physiological relevance of this enzyme, a model of mice deficient in cellular glutathione peroxidase (GSHPx-1), the major isoform of glutathione peroxidase ubiquitously expressed in all types of cells, was generated by gene-targeting technology. Mice deficient in this enzyme were apparently healthy and fertile and showed no increased sensitivity to hyperoxia. Their tissues exhibited neither a retarded rate in consuming extracellular hydrogen peroxide nor an increased content of protein carbonyl groups and lipid peroxidation compared with those of wild-type mice. However, platelets from GSHPx-1-deficient mice incubated with arachidonic acid generated less 12-hydroxyeicosatetraenoic acid and more polar products relative to control platelets at a higher concentration of arachidonic acid, presumably reflecting a decreased ability to reduce the 12-hydroperoxyeicosatetraenoic acid intermediate. These results suggest that the contribution of GSHPx-1 to the cellular antioxidant mechanism under normal animal development and physiological conditions and to the pulmonary defense against hyperoxic insult is very limited. Nevertheless, the potential antioxidant role of this enzyme in protecting cells and animals against the pathogenic effect of reactive oxygen species in other disorders remains to be defined. The knockout mouse model described in this report will also provide a new tool for future study to distinguish the physiological role of this enzyme from other selenium-containing proteins in mammals under normal and disease states.     

Chicken extract stimulates haemoglobin restoration in iron deficient rats

Chicken essence is widely used as a traditional remedy for several ailments including anaemia. To test this claim for objective evidence, a series of experiments was carried out in anaemic rats by supplementing iron deficient diets with either liquid or lyophilised essence, which contains mainly protein and peptides (83 mg/ml) and free amino acids (3.1 mg/ml), very little iron (1 microgram/ml), and no fat. Haemoglobin returned to normal significantly more rapidly in rats supplemented with ad libitum liquid BEC over a period of up to 27 days compared with controls fed only water in addition to the ad libitum iron deficient diet. Haemoglobin was also significantly increased after 1 week in animals fed ad libitum diets supplemented with lyophilised chicken essence than with controls fed the unsupplemented diet. The effect was greater with supplementation at the level of 0.2% than at 1% lyophilised essence. The results indicate that the effects were mediated by increased appetite and by enhanced availability of food iron. These studies provide objective evidence for the traditional belief that chicken essence remedies anaemia.     

DNA-enhanced peroxidase activity of a DNA-aptamer-hemin complex

BACKGROUND: In vitro selection (SELEX) previously identified short single-stranded DNAs that specifically bound N-methylmesoporphyrin IX (NMM), a stable transition-state analogue for porphyrin-metallation reactions. Interestingly, iron(III)-protoporphyrin (hemin) was a good competitive inhibitor for the DNA-catalyzed metallation reaction, and appeared to bind strongly to the NMM-binding DNA aptamers. We investigated the peroxidase activity of the aptamer-hemin complexes to see if the DNA component of the complex, like the apoenzymes in protein peroxidases, could enhance the low intrinsic peroxidatic activity of hemin. RESULTS: Two porphyrin-binding DNA aptamers bound hemin with submicromolar affinity. The aptamer-hemin complexes had significantly higher peroxidase activity than hemin alone, under physiological conditions. The Vobs of the PS2.M-hemin complex was 250 times greater than that of hemin alone, and significantly superior to a previously reported hemin-catalytic-antibody complex. Preliminary spectroscopic evidence suggests the coordination of the hemin iron in the complex changes, such that the complex more closely resembles horseradish peroxidase and other heme proteins rather than hemin. CONCLUSIONS: A new class of catalytic activity for nucleic acids is reported. The aptamer-hemin complexes described are novel DNA enzymes and their study will help elucidate the structural and functional requirements of peroxidase enzymes in general and the ways that a nucleic acid 'apoenzyme' might work to enhance the intrinsic peroxidatic ability of hemin. These aptamer-hemin complexes could be regarded as prototypes for redox-catalyzing ribozymes in a primordial 'RNA world'.     

Thyroid gland function in neonates with transient hyperthyrotrophinemia from a region of slight iodine deficiency

The aim of this study was to evaluate thyroid gland function in neonates and babies with transient hyperthyrotrophinemia during the first twelve months of life in an attempt to establish the causes of this condition in neonates born in a region of slight iodine deficiency. Thirty-eight newborns were screened. Clinical observations and measurements of serum T3, T4, TSH levels as well as urinary iodine were conducted for one year (at the age of 2 weeks, 3-4 months, and after one year of life). The screened children showed significantly higher values of T4 (p < 0.001) in comparison with the reference value in successive follow-up examinations. High T4 values may result from an increased TBG concentration in serum, and its level should be determined in the analysed material. Other hormonal values normalized after the second weak of life. Iodine deficiency was found in 80% of the children. Our assessments concerning the causes of transient hyperthyrotrophinemia conform with previous findings. It was established that the most common causes are iodine deficiency and maternal thyroid disease. None of the screened children had goitre somatic anomalies or delayed psychomotoric development did not appear more frequently than in the general pediatric population.     

Nitric oxide enhances thyroid peroxidase activity in primary human thyrocytes

Recent studies have demonstrated the production of the multi-functional messenger molecule nitric oxide (NO) by the thyroid gland. To examine a possible role for NO in thyroid function, we studied the acute and chronic effect of NO donors on thyroid peroxidase (TPO) activity in monolayer cultures of primary human thyrocytes, using a colorimetric assay technique. The presence of either S-nitrosoglutathione (GS-NO) or sodium nitroprusside (SNP) (10(-6)-10(-4) M) at the time of the assay caused a significant increase in TPO activity. Pre-incubation of thyrocytes with 10(-5) M GS-NO for 3 days had no effect on the level of TPO activity when the assay was performed in the absence of NO donors. However, GS-NO pre-incubation significantly enhanced the acute stimulatory effect of GS-NO and SNP on TPO activity. These results suggest a possible role for NO in the regulation of TPO activity and thus thyroid hormone synthesis.     

Glutathione peroxidase activity and glutathione concentration in genetically dystrophic mice

The case of a 6-year-old boy is reported who was delivered out of a frontal position by application of the vacuum cup to the left part of the forehead. His left eye exhibits a pronounced somewhat irregular astigmatism. There are some vertical descemet tears. Because of the application of the vacuum pump close to the left bulbus, an increase of the tissue tension and consequently a deformation of the bulbus occurred by the vacuum in the bulbar region. The described lesions parallel the vertical descement tears which occur in forceps-delivery. Owing to the greater horizontal corneal diameter, a more pronounced extension of the cornea in the horizontal meridian occurs, which would explain the vertical course of the descemet tears. The obstetric contraindication for the application of the vacuum extraction in case of a frontal or facial position of the fetus is justified also from the ophthalmological point of view because of the risk of occurrence of irreversible corneal lesions.