Antagonistic Interaction of Selenium and Cadmium in Human Hepatic Cells Through Selenoproteins.

Cadmium (Cd) is a highly toxic heavy metal for humans and animals, which is associated with acute hepatotoxicity. Selenium (Se) confers protection against Cd-induced toxicity in cells, diminishing the levels of ROS and increasing the activity of antioxidant selenoproteins such as glutathione peroxidase (GPx). The aim of this study was to evaluate the antagonistic effect of selenomethionine (SeMet) against Cd toxicity in HepG2 cells, through the modulation of selenoproteins. To this end, the cells were cultured in the presence of 100 µM SeMet and 5 µM, 15 µM, and 25 µM CdCl2 and a combination of both species for 24 h. At the end of the experiment, cell viability was determined by MTT assay. The total metal content of Cd and Se was analyzed by triple-quadrupole inductively coupled plasma-mass spectrometry (ICP-QqQ-MS). To quantify the concentration of three selenoproteins [GPx, selenoprotein P (SELENOP), and selenoalbumin (SeAlb)] and selenometabolites, an analytical methodology based on column switching and a species-unspecific isotopic dilution approach using two-dimensional size exclusion and affinity chromatography coupled to ICP-QqQ-MS was applied. The co-exposure of SeMet and Cd in HepG2 cells enhanced the cell viability and diminished the Cd accumulation in cells. Se supplementation increased the levels of selenometabolites, GPx, SELENOP, and SeAlb; however, the presence of Cd resulted in a significant diminution of selenometabolites and SELENOP. These results suggested that SeMet may affect the accumulation of Cd in cells, as well as the suppression of selenoprotein synthesis induced by Cd.

Assessing the viability of recovered phosphorus from eutrophicated aquatic ecosystems as a liquid fertilizer.

One of the most important worldwide environmental challenges is the alteration of the biogeochemical cycle of phosphorus (P). P is globally exported from terrestrial to aquatic ecosystems, causing the eutrophication of the receiving waters. In this context, magnetic microparticles (MPs) have been recently proposed for trapping P in natural eutrophicated ecosystems, as well as in treated wastewaters. The main advantage of using MPs is that both P and MPs can be recovered from the treated water. Thus, the working hypothesis of the present study is that P can be desorbed from P-loaded MPs and recovered P can be later used as a fertilizer. To test this hypothesis, the best working conditions for desorbing P from P-loaded MPs were identified; then, an experiment with different plant nutrient solutions (neutralized solutions containing recovered P and an unfertilized control) was carried out with three different plant species: Ocimum basilicum L., Cucumis sativus L. and Cucumis melo L. Finally, germination, height, root and shoot biomass and P concentration in root and shoot were compared among treatments. Our results show that the best conditions for P desorption from P-loaded MPs occurred when using 0.1 M NH4OH and using H3PO4 for neutralizing pH. The greenhouse fertirrigation pot experiment showed that the neutralized solution containing desorbed P from P-loaded MPs can be used as a liquid fertilizer, since its combination with macro and microelements significantly increased plant height, growth rate, shoot and root biomass and shoot and root P concentration. As a result, MPs can be proposed to be used for counteracting the widespread and coupled problems of the exhaustion of the P reserves and the eutrophication of aquatic ecosystems.

Transitioning from conformal radiotherapy to intensity-modulated radiotherapy after radical prostatectomy: Clinical benefit, oncologic outcomes and incidence of gastrointestinal and urinary toxicities.

AIM: The purpose of this study was to review genitourinary (GU) and gastrointestinal (GI) toxicity associated with high-dose radiotherapy (RT) delivered with 3-dimensional conformal radiotherapy (3D-CRT) and intensity-modulated radiotherapy (IMRT) or volumetric arc therapy (VMAT) following radical prostatectomy (RP). BACKGROUND: RP is a therapeutic option for the management of prostate cancer (PrCa). When assessing postoperative RT techniques for PrCa, the published literature focuses on patients treated with 2-dimensional conventional methods without reflecting the implementation of 3D-CRT, IMRT, or VMAT. MATERIALS AND METHODS: A total of 83 patients were included in this analysis; 30 patients received 3D-CRT, and 53 patients received IMRT/VMAT. Acute and late symptoms of the GU and lower GI tract were retrospectively graded according to the Radiation Therapy Oncology Group and the European Organization for Research and Treatment of Cancer radiation toxicity grading systems. The relapse failure-free rate and overall survival were also evaluated. RESULTS: The rate of acute GU toxicity was 9.4% vs. 13.3% for the IMRT/VMAT and 3D-CRT groups (p = 0.583). The 5-year actuarial rates of late GI toxicity for IMRT/VMAT and 3D-CRT treatments were 1.9% and 6.7%, respectively. The rate of late GU toxicity for the IMRT/VMAT and 3D-CRT treatment groups was 7.5% and 16.6%, respectively (p = 0.199). We found no association between acute or late toxicity and the RT technique in univariate and multivariate analyses. CONCLUSION: Postprostatectomy IMRT/VMAT and 3D-CRT achieved similar morbidity and cancer control outcomes. The clinical benefit of highly conformal techniques in this setting is unclear although formal analysis is needed.

Locoregional hyperthermia of deep-seated tumours applied with capacitive and radiative systems: a simulation study.

BACKGROUND: Locoregional hyperthermia is applied to deep-seated tumours in the pelvic region. Two very different heating techniques are often applied: capacitive and radiative heating. In this paper, numerical simulations are applied to compare the performance of both techniques in heating of deep-seated tumours. METHODS: Phantom simulations were performed for small (30 × 20 × 50 cm3) and large (45 × 30 × 50 cm3), homogeneous fatless and inhomogeneous fat-muscle, tissue-equivalent phantoms with a central or eccentric target region. Radiative heating was simulated with the 70 MHz AMC-4 system and capacitive heating was simulated at 13.56 MHz. Simulations were performed for small fatless, small (i.e. fat layer typically <2 cm) and large (i.e. fat layer typically >3 cm) patients with cervix, prostate, bladder and rectum cancer. Temperature distributions were simulated using constant hyperthermic-level perfusion values with tissue constraints of 44 °C and compared for both heating techniques. RESULTS: For the small homogeneous phantom, similar target heating was predicted with radiative and capacitive heating. For the large homogeneous phantom, most effective target heating was predicted with capacitive heating. For inhomogeneous phantoms, hot spots in the fat layer limit adequate capacitive heating, and simulated target temperatures with radiative heating were 2-4 °C higher. Patient simulations predicted therapeutic target temperatures with capacitive heating for fatless patients, but radiative heating was more robust for all tumour sites and patient sizes, yielding target temperatures 1-3 °C higher than those predicted for capacitive heating. CONCLUSION: Generally, radiative locoregional heating yields more favourable simulated temperature distributions for deep-seated pelvic tumours, compared with capacitive heating. Therapeutic temperatures are predicted for capacitive heating in patients with (almost) no fat.

Biological interactions between mercury and selenium in distribution and detoxification processes in mice under controlled exposure. Effects on selenoprotein.

Antagonistic interactions between mercury (Hg) and selenium (Se), were evaluated in mouse (Mus musculus), as a mammalian model, in a series of controlled exposure experiments. The beneficial effect of Se against Hg toxicity involves a variety of biochemical and toxicological processes that have not been clarified yet. For this purpose, a metallomic workflow based on the use of size-exclusion chromatography (SEC) with inductively coupled plasma mass spectrometry (ICP-MS) detection was complemented with the speciation of selenoproteins and low molecular mass selenium species in serum and liver cytosolic extracts using a multidimensional approach based on SEC-AF-HPLC-ICPMS, using species-unspecific isotope dilution (SUID)-ICP-MS for selenium quantification. The results showed potential interactions between Hg/Se in organs and serum related to accumulation and detoxification processes, in addition to the effects of mercury on selenoproteins in hepatic cytosolic extracts and bloodstream when both elements are administrated at the same time. These results provide information about elements distribution, interactions and homeostasis and reveal the potential of metallomic approaches in exposure experiments.

Valorization of phosphogypsum waste as asphaltic bitumen modifier.

The accumulation of phosphogypsum waste from the fertilizer industries, which remain in regulated stacks occupying considerable land resources, is causing significant environment problems worldwide. In that sense, the scientific community is being pressured to find alternative ways for their disposal. In this research, we propose a novel application for phosphogypsum waste, as a modifier of bitumen for flexible road pavements. Viscous flow tests carried out on bitumen modified with a phosphogypsum waste and doped with sulfuric acid demonstrated an extraordinary increase in viscosity, at 60°C, when compared to a counterpart sample which had been modified with gypsum, the main component of phosphogypsum. Similarly, a significant improvement in the viscoelastic response of the resulting material at high temperatures was also found. FTIR (Fourier transform infrared spectroscopy) scans provided evidences of the existence of chemical reactions involving phosphorus, as revealed by a new absorption band from 1060 to 1180cm(-1), related to COP vibrations. This result points at phosphorus contained in the phosphogypsum impurities to be the actual "modifying" substance. Furthermore, no COP band was observed in the absence of sulfuric acid, which seems to be the "promoting" agent of this type of bond.

Cadmium toxicity in Mus musculus mice based on a metallomic study. Antagonistic interaction between Se and Cd in the bloodstream.

Cadmium (Cd) is an important inorganic toxicant in the environment which impacts on human health. A metallomic approach based on size-exclusion chromatography (SEC) coupled to inductively coupled plasma-mass spectrometry (ICP-MS) and multidimensional chromatography separation based on SEC coupled to affinity chromatography 2D-SEC-AF-ICP-MS have been applied to achieve a better understanding of the function, detoxification processes and regulation of metals in mice (Mus musculus) under controlled exposure to both Cd and Cd plus (77)Se. Isotopic dilution analysis (IDA) was performed to quantify selenium containing proteins in mice plasma with ICP-qMS as a multielemental detector. Additionally, isotope pattern deconvolution (IPD) was applied to study the fate of enriched (77)selenite in mice subjected to cadmium exposure and the effect of selenoprotein production in plasma. Moreover, the affinity of Cd for SeP in plasma of mice was corroborated using anion exchange chromatography (AEC) after AF separation and identified by organic mass spectrometry. This work illustrates the high reliability of the integrated use of inorganic and organic mass spectrometry to get a metallomic approximation, which provides a good alternative to gain deep insight into the fate of elements in exposed organisms, providing information about metal trafficking, interactions and homeostasis.

Controle de qualidade e análise sensorial em voluntários de xampu esfoliante com extrato hidroalcoólico de Capsicum frutescens L. (Solanaceae) / Quality control and sensory acceptability of exfoliating shampoo containing hydroethanolic extract of Capsicum frutescens L. (Solanaceae) peppers.

A fitocosmética é um setor em nítido crescimento por se ter verificado reais vantagens na incorporação de produtos vegetais em relação a certos produtos sintéticos em produtos cosméticos. O proposto trabalho foi desenvolvido enfocando-se o uso de xampu com a utilização de microesferas de biopolímero e extrato hidroalcoólico de Capsicum frutescens L., mais popularmente conhecida como pimenta-malagueta, para o tratamento de queda capilar simples (alopecia androgênica). As análises físico-químicas envolveram determinação de cinzas totais, pH, identificação de capsaicina e densidade do xampu, entre outros. Considerando-se os diversos aspectos exigidos para a garantia da qualidade do material botânico, que englobam não somente os aspectos físico-químicos, mas também o microbiológico, realizou-se, tanto no extrato quanto no produto final, a contagem total de micro-organismos, pesquisa de Salmonella sp e Escherichia coli, Staphylococcus aureus e Pseudomonas aeruginosa. Foram realizadas análises sensoriais em voluntários, visando avaliar a qualidade sensorial do produto. Os voluntários avaliaram a amostra segundo questionário aplicado. Para as propriedades avaliadas, as microesferas esfoliantes incorporadas ao xampu mostraram-se ser bem aceitas pelos voluntários, principalmente devido à sensação transmitida pela esfoliação, fazendo com que o extrato hidroalcoólico promova uma maior oxigenação local.

Phytocosmetics is a sector of the market in rapid growth, since real advantages have been demonstrated in the incorporation of plant products in cosmetics, in place of certain synthetic products. The focus of this study was on the use of shampoo containing microspheres of biopolymer and a hydroalcoholic extract of the fruit of Capsicum frutescens L., more commonly known as chili pepper, for the treatment of hair loss (androgenic alopecia). The physical and chemical analysis included the determination of total ash, pH and density of the shampoo and the identification of capsaicin. Considering the various requirements for quality assurance of the plant material, encompassing not only physicochemical but also microbiological criteria, both the extract and the final product were subjected to a total count of microorganisms and tests for Salmonella sp, Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa. The sensory quality of the product was assessed by a panel of volunteers, who rated the sample of shampoo in response to a questionnaire. For the properties assessed, the exfoliating microspheres incorporated into the shampoo proved to be well tolerated by the volunteers, mainly on account of the feeling conveyed by exfoliation, causing the hydroalcoholic extract to promote greater local oxygenation.

Identification and characterization of new functional truncated variants of somatostatin receptor subtype 5 in rodents.

Somatostatin and cortistatin exert multiple biological actions through five receptors (sst1-5); however, not all their effects can be explained by activation of sst1-5. Indeed, we recently identified novel truncated but functional human sst5-variants, present in normal and tumoral tissues. In this study, we identified and characterized three novel truncated sst5 variants in mice and one in rats displaying different numbers of transmembrane-domains [TMD; sst5TMD4, sst5TMD2, sst5TMD1 (mouse-variants) and sst5TMD1 (rat-variant)]. These sst5 variants: (1) are functional to mediate ligand-selective-induced variations in [Ca(2+)]i and cAMP despite being truncated; (2) display preferential intracellular distribution; (3) mostly share full-length sst5 tissue distribution, but exhibit unique differences; (4) are differentially regulated by changes in hormonal/metabolic environment in a tissue- (e.g., central vs. systemic) and ligand-dependent manner. Altogether, our results demonstrate the existence of new truncated sst5-variants with unique ligand-selective signaling properties, which could contribute to further understanding the complex, distinct pathophysiological roles of somatostatin and cortistatin.

[Therapeutic recommendations in the oncohematological patient]. / Recomendaciones nutricionales en el paciente oncohematológico.

More than 10 million new cancer cases are detected each year worldwide, 95% of which are caused by predisposing factors, and of those, more than one third are linked to dietary factors as the main cause. The ability of maintaining an adequate nutritional status in oncohematologic patients is a common problem since the disease itself and the therapy may lead to a protein-caloric hyponutrition state that influence their quality of life and survival. For that reason, in this section we will focus on the prevalence and etiology of hyponutrition in oncologic patients, assessing the possible causes related with the tumor itself, with the patient and with administered therapies. We will also discuss performing a correct nutritional assessment in this type of patients and thus determining the main effects derived from hyponutrition status; finally, we will discuss the objectives of nutritional support and the best nutritional plan that will have to be adjusted to each patient.

Recomendaciones nutricionales en el paciente oncohematológico / Therapeutic recommendations in the oncohematological patient

Globalmente, en el mundo, se producen cada año más de 10 millones de casos nuevos de cáncer, el 95% de los cuales están causados por factores ambientales y, de ellos, más de la tercera parte están ligados a factores dietéticos como causa principal. La capacidad para mantener un estado nutricional adecuado en el paciente oncohematológico es un problema común; ya que tanto el desarrollo de la propia enfermedad como su tratamiento, pueden dar lugar a un estado de desnutrición calórico proteico, que afecta a su calidad de vida y su supervivencia. Por ello en este apartado, abordaremos la prevalencia y la etiología de la desnutrición en el paciente oncológico, valorando las posibles causas relacionadas con el propio tumor, con el paciente o con los tratamientos administrados. Pero también se mencionarán las pautas para realizar una correcta evaluación nutricional en este tipo de pacientes y determinar así los principales efectos derivados del estado de desnutrición, y por último se abordarán los objetivos del soporte nutricional así como un plan nutricional óptimo que deberá ser ajustado a cada paciente (AU)

More than 10 million new cancer cases are detected each year worldwide, 95% of which are caused by predisposing factors, and of those, more than one third are linked to dietary factors as the main cause. The ability of maintaining an adequate nutritional status in oncohematologic patients is a common problem since the disease itself and the therapy may lead to a protein-caloric hyponutrition state that influence their quality of life and survival.For that reason, in this section we will focus on the prevalence and etiology of hyponutrition in oncologic patients, assessing the possible causes related with the tumor itself, with the patient and with administered therapies.We will also discuss performing a correct nutritional assessment in this type of patients and thus determining the main effects derived from hyponutrition status; finally, we will discuss the objectives of nutritional support and the best nutritional plan that will have to be adjusted to each patient (AU)

Carbohydrate supplementation during intense exercise and the immune response of cyclists.

OBJECTIVE: To evaluate the effect of carbohydrate supplementation upon some aspects of the immune function in athletes during intense indoor cycling. METHODS: Twelve male athletes cycled for 20 min at a velocity corresponding to 90% of that obtained at the anaerobic threshold and rested for 20 min. This protocol was repeated six times. The athletes received, during the trial, water ad libitum, or a solution of carbohydrate (95% glucose polymers and 5% fructose) at 10% (w/v), 1 g kg h every 20 min, starting at the 10th minute of the first exercise period, plus extra water ad libitum. RESULTS: Exercise induced a reduction in peripheral blood mononuclear cell proliferation (37%) as well as in the production of cytokines by cultured cells (interleukin-1 (IL-1), interleukin-2 (IL-2), tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma), by 37%, 35%, 26% and 16%, respectively). All of these changes were prevented by the ingestion of a carbohydrate drink by the athletes, except that in IFN-gamma production, which was equally decreased (17%) after the second trial. The concentration of plasma glutamine, an important fuel for immune cells, was decreased in the placebo group but maintained in the group that received carbohydrate. CONCLUSION: Carbohydrate supplementation affects positively the immune response of cyclists by avoiding or minimizing changes in plasma glutamine concentration.

Tratamiento quirúrgico de las fracturas patológicas metastásicas del fémur proximal / Surgical treatment of metastatic pathological fractures of the proximal femur

Objetivo: El objetivo fue analizar retrospectivamente los resultados del tratamiento quirúrgico de 31 fracturas patológicas del fémur proximal por metástasis óseas, tratadas en el Hospital Universitario de Getafe de Madrid, desde 1991 a 2000. Pacientes y método: El origen primario fue en 12 pacientes un carcinoma de mama, en 7 de pulmón, en 5 de próstata, en 3 de riñón y hubo 4 de otros tipos. Las fracturas se localizaron en el cuello del fémur en 13 pacientes, en la región intertrocantérea en 7 y en la subtrocantérea en 11.Se realizó enclavado intramedular cerrojado en 18 pacientes y artroplastia cementada de cadera en 13 (7 parciales y 6 totales). Se valoró: el dolor residual durante la evolución postoperatoria, la mejoría funcional para la marcha y para los cuidados del paciente, el tiempo de supervivencia a la cirugía y la evolución postoperatoria de la lesión. Resultados: La calidad de vida mejoró en todos los pacientes, al disminuir la intensidad del dolor. Se consiguió la movilización en todos los pacientes y 19 recuperaron la capacidad de marcha. En 20 casos el tamaño de la lesión aumentó. La supervivencia media fue de 6,6 meses, excluyendo 4 pacientes fallecidos en el primer mes postoperatorio. Conclusión: Es posible mejorar la calidad de vida de los pacientes con fracturas patológicas metástasicas del fémur proximal. (AU)

Amphibian melanotrope subpopulations respond differentially to hypothalamic secreto-inhibitors.

The melanotrope population of the frog intermediate lobe consists of two subtypes of cells, referred to as high-(HD) and low-density (LD) melanotrope cells, which differ markedly in their basal morphofunctional features as well as their in vitro response to hypothalamic factors, such as the stimulator thyrotropin-releasing hormone (TRH) and the inhibitor dopamine. In this study, we have investigated whether other major hypothalamic regulators of the release of alpha-melanocyte-stimulating hormone (alpha-MSH), such as gamma-aminobutyric acid (GABA) and neuropeptide Y (NPY), also differentially regulate frog melanotrope subpopulations. Our results show that in LD cells, both factors markedly inhibited proopiomelanocortin (POMC) mRNA accumulation and alpha-MSH secretion. In contrast, the secretory and biosynthetic activity of HD cells was not modified by GABA. NPY inhibited POMC transcript accumulation and tended to reduce alpha-MSH secretion in HD cells, yet these effects were less pronounced than those evoked in LD cells. In addition, GABA and NPY inhibited the KCl-induced rise in cytosolic free calcium levels in both subpopulations. Taken together, these results further indicate that frog melanotrope subpopulations are differentially regulated by the hypothalamus and strongly suggest that the intensity of such regulation is directly related to the activity of the cell subset. Thus, the LD subpopulation represents a highly responsive cell subset which is regulated by multiple neuroendocrine factors (TRH, dopamine, GABA and NPY), whereas the hormone storage HD subpopulation shows a moderate response to single stimulatory (TRH) and inhibitory (NPY) inputs.

Phlebotomine sandflies and leishmaniasis risks in Colombian coffee plantations under two systems of cultivation.

The phlebotomine sandfly fauna of traditional (shaded) and intensified (unshaded) coffee plantations in Colombia was sampled by a variety of methods and the species composition and density under the two systems compared. Twenty species of Lutzomyia sandflies (Diptera: Psychodidae: Phlebotominae) were collected, of which eight were found only in the 'Coffee Axis' ('Eje Cafetero') of the departments of Caldas, Risaralda and Quindio, six were exclusive to the department of Norte de Santander and six occurred in both regions. Four species were collected only in traditional plantations and two exclusively in intensified ones. At least 13 species occurred in both plantation types. Fifteen species are opportunistic man-biters and eight are suspected vectors of leishmaniasis caused by Le. braziliensis, Le. panamensis or Le. mexicana. Seven species were collected inside houses and may be involved in intradomiciliary transmission of Leishmania. The dominant species in Norte de Santander was Lu. spinicrassa, which made up 93.8% of all the sandflies collected in this department. This species was absent from the Eje Cafetero and a number of others among the 15 recorded there might be responsible for Leishmania transmission in this region, including Lu. trapidoi, Lu. yuilli, Lu. gomezi, L. hartmanni and Lu. ovallesi. Sandfly population densities were significantly higher in traditional plantations than in intensified ones. Residents of traditional plantations were able to describe sandflies in significantly more detail than those of intensified plantations, based on seven basic characteristics related to the appearance and biting behaviour of the insects.

The effect of BCAA supplementation upon the immune response of triathletes.

INTRODUCTION: Intense long-duration exercise could lead to immune suppression through a decrease in the circulating level of plasma glutamine. The decrease in plasma glutamine concentration as a consequence of intense long-duration exercise was reversed, in some cases, by supplementing the diet of the athletes with branched-chain amino acids (BCAA). To better address this question, we have evaluated some blood parameters (lymphocyte proliferation, the level of plasma cytokines, plasma glutamine concentration, and in vitro production of cytokines by peripheral blood lymphocytes) before and after the São Paulo International Triathlon, as well as the incidence of symptoms of infections between the groups. METHODS: Twelve elite male triathletes of mean age 25.5 +/- 3.2 yr (ranging from 21.4 to 30.1 yr), weighing 74.16 +/- 3.9 kg, swam 1.5 km, cycled 40 km, and ran 10 km (Olympic triathlon) in the São Paulo International Triathlon held in April 1997 and April 1998. In both events, six athletes received BCAA and the others, placebo. RESULTS: Athletes from the BCAA group (BG) presented the same levels of plasma glutamine, before and after the trial, whereas those from the placebo group showed a reduction of 22.8% in plasma glutamine concentration after the competition. Changes in the proliferative response of peripheral blood lymphocytes were accompanied by a reduction in IL-1 production after exercise (22.2%), which was reversed by BCAA supplementation (20.3%), without changes in IL-2 production. DISCUSSION: The data obtained show that BCAA supplementation can reverse the reduction in serum glutamine concentration observed after prolonged intense exercise such as an Olympic triathlon. The decrease in plasma glutamine concentration is paralleled by an increased incidence of symptoms of infections that results in augmented proliferative response of lymphocytes cultivated in the absence of mitogens. The prevention of the lowering of plasma glutamine concentration allows an increased response of lymphocytes to ConA and LPS, as well as an increased production of IL-1 and 2, TNF-alpha, and IFN-gamma, possibly linked to the lower incidence of symptoms of infection (33.84%) reported by the supplemented athletes.

Interactions between ascorbyl free radical and coenzyme Q at the plasma membrane.

A role for coenzyme Q in the stabilization of extracellular ascorbate by intact cells has been recently recognized. The aim of this work was to study the interactions between reduced ubiquinone in the plasma membrane and the ascorbyl free radical, as an approach to understand ubiquinone-mediated ascorbate stabilization at the cell surface. K-562 cells stabilized ascorbate and decreased the steady-state levels of the semiascorbyl radical. The ability of cells to reduce ascorbyl free radical was inhibited by the quinone analogs capsaicin and chloroquine and stimulated by supplementing cells with coenzyme Q10. Purified plasma membranes also reduced ascorbyl free radical in the presence of NADH. Free-radical reduction was not observed in quinone-depleted plasma membranes, but restored after its reconstitution with coenzyme Q10. Addition of reduced coenzyme Q10 to depleted membranes allowed them to reduce the signal of the ascorbyl free radical without NADH incubation and the addition of an extra amount of purified plasma membrane quinone reductase further stimulated this activity. Reduction was abolished by treatment with the reductase inhibitor p-hydroximercuribenzoate and by blocking surface glycoconjugates with the lectin wheat germ agglutinin, which supports the participation of transmembrane electron flow. The activity showed saturation kinetics by NADH and coenzyme Q, but not by the ascorbyl free radical in the range of concentrations used. Our results support that reduction of ascorbyl free radicals at the cell surface involves coenzyme Q reduction by NADH and the membrane-mediated reduction of ascorbyl free radical.

Protective role of ubiquinone in vitamin E and selenium-deficient plasma membranes.

We have studied the effects of dietary depletion of vitamin E and selenium on endogenous ubiquinone-dependent antioxidant system. Deficiency induced an increase in both coenzyme Q9 and Q10 in liver tissue, reaching a maximum between 4 and 7 weeks of deficient diet consumption. Cytochrome b5 reductase polypeptide was also enriched in membranes after 5 weeks of deficient diet consumption. Substantial DT-diaphorase activity was found in deficient, but not in control plasma membranes. Deficient membranes were very sensitive to lipid peroxidation, although a great protection was observed after incubation with NAD(P)H. Our results show that liver cells can boost endogenous ubiquinone-dependent protective mechanisms in response to deficiency in vitamin E and selenium.

Vitamin E and selenium deficiency induces expression of the ubiquinone-dependent antioxidant system at the plasma membrane.

We have used a model of dietary deficiency that leads to a chronic oxidative stress to evaluate responses that are adaptations invoked to boost cellular defense systems. Long-Evans hooded rats were fed with a diet lacking vitamin E (E) and selenium (Se) for 7 wk from weaning leading to animals deficient in both nutrients (-E -Se). In the absence of an electron donor, liver plasma membranes from these rats were more sensitive to lipid peroxidation, although they contained 40% greater amounts of ubiquinone than the plasma membranes from rats consuming diets with sufficient vitamin E and Se (+E +Se). The incubation of plasma membranes with NAD(P)H resulted in protection against peroxidation, and this effect was more pronounced in -E -Se membranes. Deficiency was accompanied by a twofold increase in redox activities associated with trans plasma membrane electron transport such as ubiquinone reductase and ascorbate free radical reductase. Staining with a polyclonal antibody against pig liver cytochrome b5 reductase, which acts as one ubiquinone reductase in the plasma membrane, showed an increased expression of the enzyme in membranes from -E -Se rats. Little DT-diaphorase activity was measured in +E +Se plasma membranes, but this activity was dramatically increased in -E -Se plasma membranes. No such increase was found in liver cytosols, which contained elevated activity of calcium-independent phospholipase A2. Thus, ubiquinone-dependent antioxidant protection in +E +Se plasma membranes is based primarily on NADH-cytochrome b5 reductase, whereas additional protection needed in -E -Se plasma membranes is supported by the increase of ubiquinone levels, increased expression of the cytochrome b5 reductase, and translocation of soluble DT-diaphorase to the plasma membrane. Our results indicate that, in the absence of vitamin E and Se, enhancement of ubiquinone-dependent reductase systems can fulfill the membrane antioxidant protection.

Antifibroproliferative effect of tenidap in chronic antigen-induced arthritis.

OBJECTIVE: To determine whether tenidap regulates extracellular matrix metabolism in chronic arthritis. METHODS: Antigen arthritis was induced in the knees of 30 rabbits. Animals were distributed into 3 groups: untreated, tenidap-treated, and diclofenac-treated rabbits. Three weeks after disease induction, synovial membranes were extracted and processed for histopathologic examination and detection of type I collagen (CI) and fibronectin (FN) by immunoperoxidase. Simultaneously, we analyzed the in vitro effect of tenidap on healthy synovial cell (SC) proliferation, FN expression and synthesis, and expression of transforming growth factor beta1 (TGFbeta1) messenger RNA. RESULTS: Untreated animals showed synovial lining hyperplasia, cellular infiltration at the sublining, and increased deposition of matrix proteins. These findings were not apparent in tenidap-treated rabbits, where CI and FN had the same distribution as in healthy synovial membranes. In vitro, tenidap inhibited SC proliferation (> or =25 microM) and down-regulated the expression and synthesis of FN in a dose-dependent manner (> or =1 microM). This antifibrotic effect was associated with a reduction of TGFbeta1 message. CONCLUSION: Tenidap down-regulates the fibroproliferative changes typical of chronic arthritis, an effect that fits the profile of a disease-modifying agent for rheumatoid arthritis.