Presence or severity of Hashimoto's thyroiditis does not influence basal calcitonin levels: observations from CROHT biobank.

PURPOSE: The influence of Hashimoto's thyroiditis (HT) on calcitonin (Ct) production is unresolved question. The aim of this study was to explore if basal Ct levels are influenced by the presence/severity of HT or correlated with clinical phenotypes of HT patients. METHODS: We included 467 HT patients and 184 control participants, from Croatian Biobank of HT patients (CROHT), in this retrospective study. Calcitonin levels between HT patients and controls were compared using Mann-Whitney test. Ct levels between two subgroups of HT patients, divided by intake of levothyroxine (LT4) therapy, were additionally tested to take into account the illness severity. Spearman rank correlation test was used to analyze correlations between Ct levels and 14 relevant phenotypes. RESULTS: We have not detected significant differences in median Ct levels between HT patients and controls (2.2 vs 2.35 pg/mL, respectively, P = 0.717) nor in-between two subgroups of HT patients (P = 0.347). We have not detected statistically significant correlations between Ct levels and clinical phenotypes, although we identified three weak nominal correlations: negative correlation of Ct with TgAb in all HT patients (r = - 0.1, P = 0.04); negative correlation of Ct with age in subgroup of HT patients without LT4 therapy (r = - 0.13, P = 0.04); positive correlation of Ct with BSA in subgroup of HT patients on LT4 therapy (r = 0.16, P = 0.042). CONCLUSION: Our results suggest that HT patients of all disease stages preserve Ct production as healthy individuals and there is no need for Ct measurements in the absence of a nodule. Additional confirmation and clarification of observed nominal correlations are needed due to potential clinical relevance of TgAb and age-dependent Ct decrease in HT women.

Development of a valid and reliable instrument for the assessment of quality of life in parents of children with clefts.

PURPOSE: Orofacial clefts are the most common congenital malformations that affect craniofacial structures. Studies show that they have a major influence on psychological development of the patient, and on their families. A review of the literature showed a lack of specific questionnaires for children and their parents. This study investigated the impact of orofacial clefts in children on the quality of life of their parents. In addition, the results of the treatment and the quality of work of the health team members involved in this process were evaluated. MATERIALS AND METHODS: For the purpose of this study, an original questionnaire was made to analyse the effect of orofacial clefts in children who had undergone surgery on the quality of life of 73 of their parents. The questionnaire consisted of 28 simple statements, which were evaluated with a 5-degree Likert scale (from 1-fully disagree to 5-fully agree), did not require any specific additional clarification, and were easy to complete. RESULTS: Analysis of areas of the questionnaire that applied to the parents, resulted in two subscales, parental social health and child social health, which had satisfactory Cronbach's coefficients (0.907 and 0.897, respectively). However, some issues had a relatively poor coefficient of internal consistency, which justified their expulsion from the final model of the parent questionnaire. CONCLUSION: The questionnaire developed for this study comprised two subscales concerned with the social health of parents/respondents and the social health of adolescents, as perceived by the parents. It was a valid and reliable instrument, and it showed satisfactory quality of life for parents of adolescents with clefts.

Differential response of antioxidative systems of maize (Zea mays L.) roots cell walls to osmotic and heavy metal stress.

An analysis of peroxidase and ascorbate oxidase activity, phenolic content and antioxidant capacity of isolated maize root cell walls was performed in controls and plants stressed with polyethylene glycol (PEG) or heavy metals, zinc or copper. Peroxidase activity (oxidative and peroxidative) was more pronounced in the ionic than in the covalent cell wall fraction. PEG induced an increase and Zn(2+) a decrease of both ionically bound peroxidase activities. In the covalent fraction, Cu(2+) decreased oxidative and increased peroxidative activity of peroxidase. Isoelectric focusing of ionically bound proteins and activity staining for peroxidase demonstrated increased intensities and appearance of new acidic isoforms, especially in Zn(2+) and PEG treatments. Most pronounced basic isoforms (pI ~ 7.5) in controls, decreased in intensity or completely disappeared in stressed plants. Ascorbate oxidase activity was significantly increased by PEG and decreased by Zn(2+) treatments, and highly correlated with peroxidase activity. Antioxidant capacity and total phenolics content increased in heavy metal-treated and decreased in PEG-treated plants. Analysis of individual phenolic components revealed p-coumaric and ferulic acids, as the most abundant, as well as ferulic acid dimers, trimers and tetramers in the cell walls; their quantity increased under stress conditions. Results presented demonstrate the existence of diverse mechanisms of plant response to different stresses.

Alteration in basal redox state of young male soccer players after a six-month training programme.

Despite worldwide popularity of soccer, there are still insufficient data about the effects of training process on oxidative stress-induced damage, which may occur during chronic exercise. The present study aimed to determine the effects of a six-month training programme on basal redox status of young male soccer players. The study included 26 male soccer players, aged 12-13, who participated in a six-month training programme, and 26 age-matched non-athletes who were not implemented in the training process. Blood samples were collected (before and after six-month training programme) in order to measure the following oxidative stress markers: index of lipid peroxidation (measured as TBARS), nitrites (NO2-), superoxide anion radical (O2-), hydrogen peroxide (H2O2), superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) level. After six months, the levels of TBARS and NO2- were significantly increased, while the O2- and H2O2 remained unchanged. On the other hand, SOD and CAT activity increased, while GSH decreased. A carefully prepared training programme could strengthen most components of antioxidant defence systems and, except lipid peroxidation, does not promote oxidative stress in response to regular physical activity. These findings could help in the improvement of training programmes for young athletes.

Distal venous arterialization and reperfusion injury: focus on oxidative status.

In patients with unreconstructable arterial occlusive disease distal venous arterialization (DVA) seems to be a promising option in the treatment. The goals of this prospective study were to assess clinical efficiency and possible impact of DVA on tissue damage by estimating oxidative status of patients with critical limb ischemia treated with this procedure. The subjects were 60 randomized patients: 30 were undergoing DVA and 30 were treated with antiaggregation therapy. During the mean follow-up period (6.13 ± 4.32 months for DVA vs. 6.74 ± 0.5 months for antiaggregation therapy) survival (p < 0.01), limb salvage (p < 0.001), pain relief (p < 0.001) and wound healing (p < 0.001) rates were significantly different between the two groups of patients in favor of the DVA group. Ten minutes after declamping we observed a decreasing trend in the lactate level in the blood of the deep venous system (p < 0.001). Also, on postoperative day 7 digital systolic pressure and digital-brachial index were higher than before the operation (p < 0.001). In blood samples collected immediately before and successively at 1, 3, 5 and 10 min postoperatively, prooxidative status (thiobarbituric acid reactive substances, O(2)(-), H(2)O(2) and nitric oxide) and antioxidative enzymes (superoxide dismutase, catalase and glutathione reductase) were determined spectrophotometrically. Using the nonparametric Friedman test, we noted statistically nonsignificant differences (p > 0.05) in values of both prooxidative parameters and enzymes of the antioxidative defense system, before and successively at 1, 3, 5 and 10 min after operation. These results indicate that there was no statistically significant reperfusion injury after revascularization, which could have been expected after this surgical procedure, thus confirming its validity in these patients.

Comparison of the physicochemical properties of a biosimilar filgrastim with those of reference filgrastim.

Recombinant human granulocyte-colony stimulating factor (filgrastim) is a therapeutic protein used primarily to reduce incidence and duration of severe neutropenia and its associated, and serious, complications. We have developed a biosimilar filgrastim (Hospira filgrastim; Nivestim) designed to be comparable to Amgen filgrastim (Neupogen). An extensive characterization study assessed the physiochemical similarity of Hospira filgrastim to Amgen filgrastim. Both drugs were supplied in 1 ml glass, single-use, prefilled syringes (five batches of each product at 480 microg/0.5 ml and one batch of each product at 300 microg/0.5 ml). Samples were evaluated using state-of-the-art analytical methods (validated in accordance with International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use or Pharmeuropa guidelines) to determine physicochemical properties, molecular characteristics, purity and biological activity. Samples were compared after long-term storage at 2-8 degrees C and storage at 40 degrees C (stress conditions) to evaluate their degradation impurity profiles. Hospira filgrastim and Amgen filgrastim were shown to have similar physicochemical properties, molecular characteristics, purity and biological activity. No significant differences in product-related impurities were recorded between Hospira filgrastim and Amgen filgrastim following storage for 12 weeks under stress conditions. These data show that the physicochemical profile of Hospira filgrastim is similar to that of Amgen filgrastim.

Hydroquinone peroxidase activity of maize root mitochondria.

The oxidation of hydroquinone with H(2)O(2) in the presence of mitochondria isolated from maize (Zea mays L.) roots was studied. The results indicate that a reduced form of quinone may be a substrate of mitochondrial peroxidases. Specific activities in different mitochondrial isolates, the apparent K (m) for hydrogen peroxide and hydroquinone, and the influence of some known peroxidase inhibitors or effectors are presented. Zymographic assays revealed that all mitochondrial peroxidases, which were stained with 4-chloro-1-naphthol, were capable of oxidizing hydroquinone. A possible antioxidative role of hydroquinone peroxidase in H(2)O(2) scavenging within the mitochondria, in cooperation with ascorbate or coupled with mitochondrial NAD(P)H dehydrogenases, is proposed.

Superoxide dismutase, peroxidase, and germin-like protein activity in plasma membranes and apoplast of maize roots.

The analysis of plasma membranes from maize roots by native gel electrophoresis revealed the existence of Mn-containing 120 kDa and CuZn-containing 70, 40, and 15 kDa superoxide dismutase (SOD) isoform activities. Isoelectric focusing of the plasma membranes differentiated anionic SOD isoforms with a pI of about 5 and cationic SOD isoforms at pI 8.6. Solubilization of the plasma membrane proteins further separated the cationic SOD into pI 8.6, 8.2, 8.4, and 7.2 isoforms. Double staining for both SOD and peroxidase activities showed an overlap of these activities only in the case of the high-molecular-mass (ca. 120 kDa) isoforms. High-temperature treatments demonstrated that the 120 kDa isoform was active even at 100 degrees C, indicating that it was a germin-like protein with superoxide-dismutating activity, different from the peroxidase with a similar molecular mass and the lower-molecular-mass CuZn-containing superoxide dismutases. These results are compared to those obtained from whole-tissue extract and apoplastic fluid.

Superoxide synthase and dismutase activity of plasma membranes from maize roots.

Superoxide synthase and superoxide dismutase activity have been monitored in isolated maize ( Zea mays) root plasma membranes spectrophotometrically by determination of nitro-blue tetrazolium and cytochrome c reduction, respectively. Superoxide production was induced by NADH and NADPH, with similar kinetics and approaching saturation at 0.06 mM in the case of NADPH and 0.1 mM in the case of NADH, with rates of 18.6 +/- 5.0 and 21.8 +/- 7.2 nmol/min. mg of protein, respectively. These activities exhibited a broad pH optimum between pH 6.5 and 7.5. Diphenylene iodonium inhibited about 25% (10 microM DPI) and 40% (100 microM DPI) of this activity, imidazole inhibited about 20%, while KCN, a peroxidase inhibitor, did not show any significant inhibition. Superoxide-dismutating activity was shown to occur in the same isolates and depended on the quantity of plasma membrane protein present. Growth of plants on salicylic acid prior to membrane isolation induced a rise in the activity of both of the enzymes by 20-35%, suggesting their coordinated action.

Characterization of cell wall oxalate oxidase from maize roots.

Oxalate oxidase activity was detected in the cell wall fraction isolated from maize roots (Zea mays L.). The enzyme was active at acidic pH with optimal activity at pH 3.2. It was thermally extremely stable and resistant to high salt concentration, SDS and pepsin. The enzyme activity was inhibited by sulphydryl reagents 2-mercaptoethanol (2-ME), N-ethyl maleimide (NEM) and dithiotreitol (DTT), but was insensitive to EDTA, KCN and metal ions. Measurements of enzyme activity were performed using colorimetric assay of H(2)O(2), as well as polarographic detection of O(2) consumption. Maximal activity was obtained with 5 mM oxalic acid for the colorimetric method, and 10 mM oxalic acid for the polarographic method. Both methods were applicable in oxalate oxidase characterization, the polarographic method being more suitable under conditions of H(2)O(2) interaction with some of the analyzed substances.

Involvement of plasma membrane redox system in the generation of trans-root electrical potential difference in excised maize root.

Possible involvement of the plasma membrane bound redox system in the generation of the trans-root electrical potential difference (TRP) arising across 8 day old maize (Zea mays L. hybrid ZPSC704) roots was studied. Excised roots were exposed to artificial impermeable electron acceptors (potassium hexacyanoferrate III and potassium hexachloroiridate IV) in external solution, and TRP response, oxygen consumption rate, proton efflux and reduction of the electron acceptors were analyzed. The effect of hexacyanoferrate III (HCF III) was tested at three concentrations (0.1; 0.5 and 1.0 mmol/l), and hexachloroiridate IV (HCI IV) in the concentration range 10(-7)-5.10(-4) mol/l. Both electron acceptors depolarized the trans-root potential, an order of magnitude lower concentrations of hexachloroiridate producing a much more rapid depolarization of greater magnitude. The roots had a higher capacity to reduce 0.1 mmol/l hexachloroiridate than 1 mmol/l hexacyanoferrate. Also, an increased level of acidification induced by HCI IV than HCF III could be observed. The rate of oxygen consumption showed an increase of about 20% in both cases. These results prove that electron transplasma membrane transport process(es) contribute to the total trans-root electrical potential difference across an excised maize root.

Electrogenic proton pump in Nitella and the effect of calcium.

The hyperpolarisation of the membrane potential in Characeae above that of the diffusion potential is explained by the operation of the electrogenic proton pump. We studied the interaction of calcium with the functioning of the pump. The membrane potential was measured using the standard microelectrode technique. An increase in the calcium concentration resulted in depolarisation, its magnitude increasing with lower proton concentrations. Calcium-induced membrane potential changes, tested in the concentration range of 0.25 mmol/l to 25 mmol/l, were greatest at 0.25 mmol/l CaCl2 and decreased with the increasing calcium concentration. Light-induced initial changes in the membrane potential also showed a dependence on the presence of calcium in the external medium. We conclude that calcium has a role in the regulation of the proton pump in Nitella.