Where do we aspire to publish? A position paper on scientific communication in biochemistry and molecular biology

The scientific publication landscape is changing quickly, with an enormous increase in options and models. Articles can be published in a complex variety of journals that differ in their presentation format (online-only or in-print), editorial organizations that maintain them (commercial and/or society-based), editorial handling (academic or professional editors), editorial board composition (academic or professional), payment options to cover editorial costs (open access or pay-to-read), indexation, visibility, branding, and other aspects. Additionally, online submissions of non-revised versions of manuscripts prior to seeking publication in a peer-reviewed journal (a practice known as pre-printing) are a growing trend in biological sciences. In this changing landscape, researchers in biochemistry and molecular biology must re-think their priorities in terms of scientific output dissemination. The evaluation processes and institutional funding for scientific publications should also be revised accordingly. This article presents the results of discussions within the Department of Biochemistry, University of São Paulo, on this subject.

Squamous odontogenic tumor: report of a case of unusual involvement / Tumor odontogênico escamoso: relato de um caso de envolvimento incomum

ABSTRACT The squamous odontogenic tumor (SOT) is defined as a very rare benign neoplasm, locally infiltrative and can extend to neighboring structures. This study aimed to report a case of SOT unusual involvement. A female patient, 56-year-old, smoker showed mild swelling in the chin region in the portion between the left canine and right first premolar. Axial, coronal and sagittal images showed expansive hypodense lesion in the mandibular symphysis and parasymphysis. Partial removal of the lesion was performed, which led to the diagnosis of SOT. The patient presented no recurrence during 4 years and 2 months follow-up.

RESUMO O tumor odontogênico escamoso (TOE) é definido como uma neoplasia benigna rara, localmente infiltrativa, podendo estender-se a estruturas vizinhas. Objetivou-se relatar um caso de TOE de acometimento incomum. Paciente do sexo feminino, 56 anos de idade, fumante, demonstrou leve aumento de volume na região do mento na porção compreendida entre o canino esquerdo e o primeiro pré-molar direito. Imagens axial, coronal e sagital demonstraram lesão expansiva hipodensa na sínfise e na linha média mandibular. Fez-se a remoção parcial da lesão, o que levou ao diagnóstico de TOE. A paciente não apresentou recidiva durante quatro anos e dois meses de acompanhamento.

Performance of absolute CD4+ count in predicting co-infection with human T-lymphotropic virus type 1 in antiretroviral-naive HIV-infected patients.

Early identification of patients co-infected with HIV and human T-lymphotropic virus type 1 (HTLV-1) is essential to improve care, as CD4+ T-cell counts have been revealed to be an unreliable laboratory parameter to monitor HIV infection in co-infection. Unfortunately, HTLV-1 testing is not currently available in sub-Saharan Africa. We conducted this study to determine the performance of absolute CD4+ T-cell count estimation in guiding the clinical suspicion of co-infection. A cross-sectional survey was conducted in antiretroviral-naïve HIV (AN-HIV) patients attending an HIV outpatient clinic in Maputo city, Mozambique. Seven hundred and one AN-HIV patients were enrolled in the study. The prevalence of HTLV-1 co-infection was 4.5% (95% confidence interval [CI] 3.0-6.0%). Logistic regression analysis showed that CD4+ T-cell count was an independent predictor of co-infection (P value: 0.000). The performance of absolute CD4+ T-cell counts in predicting co-infection was higher in symptomatic HIV patients when compared with asymptomatic HIV patients. The best performance was achieved with the cut-off of CD4+ count of 500 cells/mm(3), which gave sensitivity, specificity, positive and negative predictive values of 54.2%, 87.2%, 24.0% and 96.2%, respectively. In conclusion, our data provide evidence that the absolute CD4+ T-cell count is of moderate accuracy in guiding the clinical suspicion of co-infection in AN-HIV and its implementation could improve the care provided to a significant number of HIV patients in Mozambique.

Surveillance of HIV drug resistance in children receiving antiretroviral therapy: a pilot study of the World Health Organization's generic protocol in Maputo, Mozambique.

Between 2007 and 2008, the Mozambique Ministry of Health conducted an assessment of human immunodeficiency virus drug resistance (HIVDR) using World Health Organization (WHO) methods in a cohort of children initiating antiretroviral therapy (ART) at the main pediatric ART referral center in Mozambique. It was shown that prior to ART initiation 5.4% of children had HIVDR that was associated with nevirapine perinatal exposure (P < .001). Twelve months after ART initiation, 77% had viral load suppression (<1000 copies/mL), exceeding the WHO target of ≥ 70%; 10.3% had HIVDR at 12 months. Baseline HIVDR (P = .04), maternal prevention of mother-to-child transmission (P = .02), and estimated days of missed medication (P = .03) predicted HIVDR at 12 months. As efforts to eliminate pediatric AIDS are intensified, implementation of ritonavir-boosted protease inhibitor regimens in children with prevention of mother-to-child transmission exposure may reduce risk of virological failure in our setting.

Loss of correlation between HIV viral load and CD4+ T-cell counts in HIV/HTLV-1 co-infection in treatment naive Mozambican patients.

Seven hundred and four HIV-1/2-positive, antiretroviral therapy (ART) naïve patients were screened for HTLV-1 infection. Antibodies to HTLV-1 were found in 32/704 (4.5%) of the patients. Each co-infected individual was matched with two HIV mono-infected patients according to World Health Organization clinical stage, age +/-5 years and gender. Key clinical and laboratory characteristics were compared between the two groups. Mono-infected and co-infected patients displayed similar clinical characteristics. However, co-infected patients had higher absolute CD4+ T-cell counts (P = 0.001), higher percentage CD4+ T-cell counts (P < 0.001) and higher CD4/CD8 ratios (P < 0.001). Although HIV plasma RNA viral loads were inversely correlated with CD4+ T-cell-counts in mono-infected patients (P < 0.0001), a correlation was not found in co-infected individuals (P = 0.11). Patients with untreated HIV and HTLV-1 co-infection show a dissociation between immunological and HIV virological markers. Current recommendations for initiating ART and chemoprophylaxis against opportunistic infections in resource-poor settings rely on more readily available CD4+ T-cell counts without viral load parameters. These guidelines are not appropriate for co-infected individuals in whom high CD4+ T-cell counts persist despite high HIV viral load states. Thus, for co-infected patients, even in resource-poor settings, HIV viral loads are likely to contribute information crucial for the appropriate timing of ART introduction.

Application of the comet assay in erythrocytes of Oreochromis niloticus (Pisces): a methodological comparison

The present study applied the comet assay to erythrocytes of Oreochromis niloticus with the aim of improving protocols to detect DNA damage in these cells, by using two distinct pHs (pH = 12.1 and pH > 13) and evaluating whether there is a correspondence between silver and ethidium bromide staining. Comets were visually examined and, the frequency of cells with and without damage was obtained, as well as the distribution of classes and scores. By using the Kruskal-Wallis test, our results revealed that pH 12.1 is more effective, although both pHs can be used. Our findings also suggest that silver staining can substitute ethidium bromide, an expensive and highly toxic stain that requires specific equipment for examination.

Utilization of solar energy in the photodegradation of gasoline in water and of oil-field-produced water.

The photo-Fenton process utilizes ferrous ions (Fe2+), hydrogen peroxide (H2O2), and ultraviolet (UV) irradiation as a source of hydroxyl radicals for the oxidation of organic matter present in aqueous effluents. The cost associated with the use of artificial irradiation sources has hindered industrial application of this process. In this work, the applicability of solar radiation for the photodegradation of raw gasoline in water has been studied. The photo-Fenton process was also applied to a real effluent, i.e., oil-field-produced water, and the experimental results demonstrate the feasibility of employing solar irradiation to degrade this complex saturated-hydrocarbon-containing system.

Leishmaniose visceral entre índios no Estado de Roraima, Brasil: aspectos clínicoepidemiológicos de casos observados no período de 1989 a 1993 / Visceral leishmaniasis among Indians of the State of Roraima, Brazil: clinical and epidemiologic aspects of the cases observed from 1989 to 1993

Descreve-se o perfil epidemiológico da leishmaniose visceral entre índios no estado de Roraima, Brasil, baseado na ocorrência de casos humanos observados e nos inquéritos caninos e entomológicos realizados no período de 1989 a 1993. Foram registrados 82 casos humanos de leishmaniose visceral em seis dos oito municípios então existentes no estado; houve predomínio de 69,5 por cento para o sexo masculino entre os casos observados. A maioria (52,4 por cento) dos casos foi entre crianças de zero a dez anos de idade. Registrou-se o índice de 10,3 por cento de infecção canina natural, entre 3.773 cães examinados em 74 localidades pesquisadas. A Lutzomyia longipalpis foi encontrada nas áreas de maior prevalência da doença, em 31 localidades diferentes. Os casos humanos, caninos e vetores estão concentrados em regiões onde predominam serras e lavrados, áreas características de ocorrência da leishmaniose visceral americana. A introdução e intensificação das atividades garimpeiras na região podem ter contribuído para a introdução da doença.

EPR detection of glutathionyl and protein-tyrosyl radicals during the interaction of peroxynitrite with macrophages (J774).

Peroxynitrite is one of the biological oxidants whose addition to cells has been shown to either activate signaling pathways or lead to cell injury, depending on cell type and oxidant concentration. The intermediacy of free radicals in these processes has been directly demonstrated only during the interaction of peroxynitrite with erythrocytes, a particular cell type, due to its high hemoglobin content. Here, we demonstrate that the addition of peroxynitrite to a macrophage cell line (J774) led to the production of glutathionyl and protein-tyrosyl radicals. The glutathionyl radical was characterized by EPR spin-trapping experiments with 5,5-dimethyl-1-pyrroline-N-oxide. Protein-tyrosyl radical formation was suggested by direct EPR spectroscopy and confirmed by EPR spin-trapping experiments with 3,5-dibromo-4-nitrosobenzenesulfonic acid and Western blot analysis of nitrated proteins in treated macrophages. Time dependence studies of free radical formation indicate that intracellular glutathione and unidentified proteins are the initial peroxynitrite targets in macrophages and that their derived radicals trigger radical chain reactions. The results are likely to be relevant to the understanding of the bioregulatory and biodamaging effects of peroxynitrite.

Role of peroxynitrite in macrophage microbicidal mechanisms in vivo revealed by protein nitration and hydroxylation.

The cytotoxins produced by phagocytic cells lacking peroxidases such as macrophages remain elusive. To elucidate macrophage microbicidal mechanisms in vivo, we compared the lesion tissue responses of resistant (C57Bl/6) and susceptible (BALB/c) mice to Leishmania amazonensis infection. This comparison demonstrated that parasite control relied on lesion macrophage activation with inducible nitric oxide synthase expression (iNOS), nitric oxide synthesis, and extensive nitration of parasites inside macrophage phagolysosomes at an early infection stage. Nitration and iNOS expression were monitored by confocal microscopy; nitric oxide synthesis was monitored by EPR. The main macrophage nitrating agent was shown to be peroxynitrite derived because parasite nitration occurred in the virtual absence of polymorphonuclear cells (monitored as peroxidase activity) and was accompanied by protein hydroxylation (monitored as 3-hydroxytyrosine levels). In vitro studies confirmed that peroxynitrite is cytotoxic to parasites whereas nitric oxide is cytostatic. The results indicate that peroxynitrite is likely to be produced close to the parasites and most of it reacts with carbon dioxide to produce carbonate radical anion and nitrogen dioxide whose concerted action leads to parasite nitration. In parallel, some peroxynitrite decomposition to the hydroxyl radical should occur due to the detection of hydroxylated proteins in the healing tissues. Consequently, peroxynitrite and derived radicals are likely to be important macrophage-derived cytotoxins.

Carbon dioxide stimulates the production of thiyl, sulfinyl, and disulfide radical anion from thiol oxidation by peroxynitrite.

Reaction of peroxynitrite with the biological ubiquitous CO(2) produces about 35% yields of two relatively strong one-electron oxidants, CO(3) and ( small middle dot)NO(2), but the remaining of peroxynitrite is isomerized to the innocuous nitrate. Partial oxidant deactivation may confound interpretation of the effects of HCO3-/CO(2) on the oxidation of targets that react with peroxynitrite by both one- and two-electron mechanisms. Thiols are example of such targets, and previous studies have reported that HCO3-/CO(2) partially inhibits GSH oxidation by peroxynitrite at pH 7.4. To differentiate the effects of HCO3-/CO(2) on two- and one-electron thiol oxidation, we monitored GSH, cysteine, and albumin oxidation by peroxynitrite at pH 5.4 and 7.4 by thiol disappearance, oxygen consumption, fast flow EPR, and EPR spin trapping. Our results demonstrate that HCO3-/CO(2) diverts thiol oxidation by peroxynitrite from two- to one-electron mechanisms particularly at neutral pH. At acid pH values, thiol oxidation to free radicals predominates even in the absence of HCO3-/CO(2). In addition to the previously characterized thiyl radicals (RS.), we also characterized radicals derived from them such as the corresponding sulfinyl (RSO.) and disulfide anion radical (RSSR.-) of both GSH and cysteine. Thiyl, RSO. and RSSR.- are reactive radicals that may contribute to the biodamaging and bioregulatory actions of peroxynitrite.

Inhibition of vascular NADH/NADPH oxidase activity by thiol reagents: lack of correlation with cellular glutathione redox status.

Vascular NAD(P)H oxidase activity contributes to oxidative stress. Thiol oxidants inhibit leukocyte NADPH oxidase. To assess the role of reactive thiols on vascular oxidase, rabbit iliac/carotid artery homogenates were incubated with distinct thiol reagents. NAD(P)H-driven enzyme activity, assessed by lucigenin (5 or 250 microM) luminescence, was nearly completely (> 97%) inhibited by the oxidant diamide (1mM) or the alkylator p-chloromercuryphenylsulfonate (pCMPS, 0.5mM). Analogous inhibition was also shown with EPR spectroscopy using DMPO as a spin trap. The oxidant dithionitrobenzoic acid (0.5mM) inhibited NADPH-driven signals by 92% but had no effect on NADH-driven signals. In contrast, the vicinal dithiol ligand phenylarsine oxide (PAO, 1 microM) induced minor nonsignificant inhibition of NADPH-driven activity, but significant stimulation of NADH-triggered signals. The alkylator N-ethyl maleimide (NEM, 0.5mM) or glutathione disulfide (GSSG, 3mM) had no effect with each substrate. Coincubation of N-acetylcysteine (NAC, 3mM) with diamide or pCMPS reversed their inhibitory effects by 30-60%, whereas NAC alone inhibited the oxidase by 52%. Incubation of intact arterial rings with the above reagents disclosed similar results, except that PAO became inhibitor and NAC stimulator of NADH-driven signals. Notably, the cell-impermeant reagent pCMPS was also inhibitory in whole rings, suggesting that reactive thiol(s) affecting oxidase activity are highly accessible. Since lack of oxidase inhibition by NEM or GSSG occurred despite significant cellular glutathione depletion, change in intracellular redox status is not sufficient to account for oxidase inhibition. Moreover, the observed differences between NADPH and NADH-driven oxidase activity point to complex or multiple enzyme forms.

In vivo metabolism of tert-butyl hydroperoxide to methyl radicals. EPR spin-trapping and DNA methylation studies.

Metabolic activation of peroxides and hydroperoxides to free radicals is associated with the tumor promoting activity of these compounds. tert-Butyl hydroperoxide (t-BOOH) metabolism has been extensively studied as a model of peroxide biotransformation. In vivo studies are limited, and the hemoglobin-thiyl radical was the only species thus far identified in the blood of treated rats. Here we further examine t-BOOH metabolism in vivo with regard to free radical and DNA adduct production. Spin-trapping experiments with phenyl-N-tert-butylnitrone (PBN) led to the detection of EPR signals in the blood, bile, and organic extracts of the liver and stomach of rats treated with t-BOOH. Analysis of these signals demonstrated that t-BOOH metabolism in vivo produces alkyl radicals, detected in the bile and organic extracts of liver and stomach, in addition to the previously identified hemoglobin-thiyl radical. To characterize the produced alkyl radicals, experiments were performed with (13)C-labeled t-BOOH and two spin traps, PBN and alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone (POBN). The latter was used because the EPR signals obtained with PBN were too weak to be unambiguous. Nevertheless, the EPR signals present in the bile of animals treated with (13)C-labeled t-BOOH and PBN or POBN were consistent with adducts of (13)C-labeled methyl radical and an unidentified alkyl radical. The latter is probably derived from lipids oxidized by the metabolically produced primary radicals, methyl and its precursor, tert-butoxyl. The presence of 8-methylguanine and 7-methylguanine in hydrolysates of DNA from liver and stomach of rats treated with t-BOOH was also examined. 8-Methylguanine, a typical product of methyl radical attack on DNA, was detectable in both the liver and stomach of treated rats. The results may be relevant to the understanding of the genotoxic properties of other peroxides, particularly of cumene hydroperoxide.

Vascular oxidant stress early after balloon injury: evidence for increased NAD(P)H oxidoreductase activity.

Available evidence for oxidative stress after angioplasty is indirect or ambiguous. We sought to characterize the pattern, time course, and possible sources of free radical generation early after arterial balloon injury. Ex vivo injury performed in arterial rings in buffer with lucigenin yielded a massive oxygen-dependent peak of luminescence that decayed exponentially and was proportional to the degree of injury. Signals for injured vs. control arteries were 207. 1 +/- 17.9 (n = 13) vs 4.1 +/- 0.7 (n = 22) cpm x 10(3)/mg/min (p <. 001). Data obtained with 0.25 mmol/l lucigenin were validated with 0. 005-0.05 mmol/l lucigenin or the novel superoxide-sensitive probe coelenterazine (5 micromol/l). Gentle removal of endothelium prior to injury scarcely affected the amount of luminescence. Lucigenin signals were amplified 5- to 20-fold by exogenous NAD(P)H, and were >85% inhibited by diphenyliodonium (DPI, a flavoenzyme inhibitor). Antagonists of several other potential free radical sources, including xanthine oxidase, nitric oxide synthase, and mitochondrial electron transport, were without effect. Overdistension of intact rabbit iliac arteries in vivo (n = 7) induced 72% fall in intracellular reduced glutathione and 68% increase in oxidized glutathione, so that GSH/GSSG ratio changed from 7.93 +/- 2.14 to 0. 81 +/- 0.16 (p <.005). There was also 28.7% loss of the glutathione pool. Further studies were performed with electron paramagnetic resonance spectroscopy. Rabbit aortas submitted to ex vivo overdistension in the presence of the spin trap DEPMPO (5-diethoxy-phosphoryl-5-methyl-1-pyrroline-N-oxide, 100 mmol/l, n = 5) showed formation of radical adduct spectra, abolished by DPI or superoxide dismutase. Computer simulation indicated a mixture of hydroxyl and carbon-centered radical adducts, likely due to decay of superoxide adduct. Electrical mobility shift assays for NF-kappaB activation were performed in nuclear protein extracts from intact or previously injured rabbit aortas. Balloon injury induced early NF-kappaB activation, which was decreased by DPI. In conclusion, our data show unambiguously that arterial injury induces an immediate profound vascular oxidative stress. Such redox imbalance is likely accounted for by activation of vessel wall NAD(P)H oxidoreductase(s), generating radical species potentially involved in tissue repair.

La residencia de Cirugía General en nuestro hospítal: Cómo se creó, qué hacemos y qué resultados obtenemos. Un análisis de doce años / The General Surgery residence in our hospital: How it was created, what we do and what results we get. A twelve year analysis

Se presenta una reseña sobre la Residencia del Servicio de Clínica Quirúrgica del HIG Dr. José Penna, de Bahía Blanca. En doce años de existencia, se han formado íntegramente o están completando su instrucción, 26 profesionales sobre 27 ingresantes; 25 varones y 2 mujeres, con una edad promedio, al ingreso de 26 años. Diecisiete residentes provinieron de la provincia de Buenos Aires (once de la zona de influencia del hospital y seis de otras regiones); 9, de otras provincias y uno de la República del Perú. La Universidad Nacional de La Plata fue la mayor proveedora de recurso humano con 18 ingresantes, distribuyéndose el resto entre las U. N. de Córdoba, Litoral, Buenos Aires y Católica, de Córdoba. Cada residente asistió, en promedio, a 728 pacientes, realizó 545 operaciones como cirujano y ayudó en más de 2.000 intervenciones, durante su período formativo. Actualmente, 15 residentes han sido absorbidos por el ámbito laboral local (8 en nuestro hospital y 7 en otros establecimientos privados). Los restantes se desempeñan como cirujanos en otros sitios del país. Se concluye que nuestro servicio, además de ser evaluado y jerarquizado por la Asociación Argentina de Cirugía, ha demostrado competencia para formar un recurso humano útil para el hospital, con una capacitación adecuada y sin dificultad de integración laboral, en nuestro medio o en otros lugares. Aún resta una mayor complementación con otras especialidades para poder dotar a nuestros residentes de un bagaje práctico más amplio y una optimización de su formación teórica.

In this paper we present a review about the Residency at the Unit of Surgical Clinic of the HIG Dr. José Penna, in Bahía Blanca. In the last twelve years, 26 professionals, out of 27 entrant students, have been trained comprehensively or are still completing their training program. The group includes 25 male students and 2 female students who, in average, are 26 years old at the time they begin the training. Seventeen residents came from Buenos Aires Province (eleven from the area of service of the hospital and six from other regions), nine from other provinces, and one from the Republic of Perú. The Universidad Nacional de La Plata was the largest supplier of human resources, with 18 entrant students, the rest came from the following universities: U. N. de Córdoba, Litoral, Buenos Aires and Católica, from Córdoba. Each resident assisted an average of 728 patients, performed 545 operations as surgeon and helped in more than 2,000 surgeries during their training period. At present, 15 residents are working at local institutions (8 at our hospital and 7 at other private facilities). The rest are practicing surgeons in other parts of the country. We conclude that our unit, in addition to the assessment and hierarchy granted by the Argentine Association of Surgery, has shown competence in the training of human resources useful for the hospital, with adequate training and with no difficulty to become part of the work market, either in our area or in other places. We still have to achieve a greater complementation with other specializations in order to offer our residents wider handson experience and better theoretical training.

Oxidation of acetaldehyde by peroxynitrite and hydrogen Peroxide/Iron(II). Production Of acetate, formate, and methyl radicals.

Production of free radicals from acetaldehyde oxidation by enzymes and cellular fractions is a well-known process. The toxic effects of acetaldehyde, however, are usually attributed to its reactions with biomolecules to produce adducts. Here, we demonstrate that hypothetical adducts produced from attack of acetaldehyde by two important biological oxidants, peroxynitrite and hydrogen peroxide, decompose to produce acetate, formate, and methyl radicals. Acetate, formate, nitrate, and nitrite were characterized and quantified by capillary electrophoresis. Radicals were detected and quantified by the EPR spectra produced in the presence of spin traps 3, 5-dibromo-4-nitrosobenzenesulfonic acid and 5,5-dimethyl-1-pyrroline N-oxide. Kinetic studies and product analysis were performed at different pHs. The results demonstrate that production of methyl radicals during oxidation of acetaldehyde by hydrogen peroxide was strictly dependent on the presence of iron(II) and occurred via two routes. One involved acetaldehyde attack by the hydroxyl radical to produce the acetyl radical that decomposes to methyl radical and carbon monoxide. The other route involved acetaldehyde attack by deprotonated hydrogen peroxide to produce a hypothetical intermediate that reductively cleaves via the action of present iron(II) to produce radicals. The latter mechanism predominates in the case of peroxynitrite, but radical formation does not require metal ions. Most of the hypothetical adduct produced from acetaldehyde and peroxynitrite (k = 680 M(-)(1) s(-)(1) at pH 7.4 and 37 degrees C) decays to nitrate and regenerates the aldehyde [Uppu, R. M., et al. (1997) Chem. Res. Toxicol. 10, 1331], but about 30% of it produces acetate, formate, and methyl radicals. Part of these oxidized products result from beta-scission and 1,2-shift reactions of the 1-hydroxyethoxyl radical which, together with nitrogen dioxide, freely diffuses from the adduct (20% yields). The results provide yet another example of the metal-independent free radical reactivity of peroxynitrite and may be relevant to the toxic effects associated with heavy drinking and diabetes.

DNA methylation by tert-butyl hydroperoxide-iron (II): a role for the transition metal ion in the production of DNA base adducts.

Metabolic degradation of both endogenous and exogenous peroxides is associated with the etiology of several diseases including cancer. Tert-butyl hydroperoxide (TBHP) has been widely employed as a model compound to study the cytotoxicity and promoting effects of organic peroxides. Recently, we reported that incubations of TBHP with iron (II) and calf thymus DNA led to generation of high yields of methyl radicals and to DNA methylation. Interestingly, DNA was methylated to products expected from both free radical and ionic mechanisms such as 8-methylguanine (C8-MeGua) and 7-methylguanine (N7-MeGua), respectively. To elucidate the mechanisms by which methyl radicals can produce different types of DNA adducts, we examined the effects of transition metal ions (iron (II), iron (III) and copper (I)) and metal ion chelators (ethylenediamine-N,N,N",N"-tetraacetate (EDTA) and desferal) on the nature and the yields of the DNA adducts produced during TBHP decomposition. The results led us to propose that a direct methyl radical attack on DNA guanine residues produces C8-MeGua whereas N7-MeGua and 3-methyladenine (N3-MeAde) are likely to be produced by attack of nucleophilic DNA centers on methyl radical generated in situ by the assistance of transition metal ions bound to DNA.

Tempo de permanência hospitalar de recém-nascidos com mais de 1.000 gramas de peso de nascimento / Time of hospital stay for newborn at the Neonatal Unit of HIAE with more of the 1.000 gram of birth weight

Foi estudado o tempo de permanência hospitalar de recém-nascidos da unidade Neonatal do HIAE, segundo o peso de nascimento e critérios de alta estabelecidos previamente. Foram calculados as médias e seus respectivos desvios-padräo, de modo a possibilitar análises individuais do comportamento de cada recém-nascido e identificar as possíveis causas de eventuais desvios

Nucleic acid alkylation by free radical metabolites of ethanol. Formation of 8-(1-hydroxyethyl)guanine and 8-(2-hydroxyethyl)guanine adducts.

Alcohol consumption is associated with an increased risk of several types of malignancy by mechanisms that remain to be elucidated. Most of the ingested ethanol is converted to acetaldehyde but the formation of free radical metabolites such as the 1-hydroxyethyl radical has been also demonstrated to occur in vitro and in vivo. Here we tested the possibility of ethanol-derived free radicals alkylating nucleic acid and nucleic acid components. Ethanol oxidation by Fenton systems has been extensively used to mimic ethanol metabolism to free radical intermediates and it was also employed in our studies. Two adducts, 8-(1-hydroxyethyl)guanine and 8-(2-hydroxyethyl)guanine, were isolated in incubations containing guanine/ethanol/hydrogen peroxide/iron(II) at pH 1 under anaerobic conditions. The adducts were produced in comparable yields and were characterized by ultraviolet absorption, mass spectrometry, and proton nuclear magnetic resonance spectroscopy. Both adducts were also produced in incubations containing DNA and RNA at pH 4 and 7. Under these conditions, the obtained yields of 8-(1-hydroxyethyl)guanine were about 10 times higher than those of 8-(2-hydroxyethyl)guanine. Higher yields of both adducts were obtained at pH 4 than at pH 7 and with RNA as compared with DNA. As expected, nucleic acid oxidation products such as 8-oxo-7,8-dihydroguanine and 8-oxo-7,8-dihydroadenine were also produced under the employed experimental conditions. Their yields tended to increase in the presence of ethanol, particularly at pH 4, suggesting that ethanol can protect oxidized bases from further degradation. Parallel spin-trapping experiments with alpha-4-pyridyl-1-oxide N-tert-butylnitrone and 3,5-dibromo-4-nitrosobenzenesulfonic acid confirmed that ethanol was oxidized to both the 1-hydroxyethyl and 2-hydroxyethyl radicals by hydrogen peroxide/iron(II) at pH 4-7 in the presence and in the absence of nucleic acids. The results demonstrate that free radical metabolites of ethanol can alkylate nucleic acids in vitro. Both the 1-hydroxyethyl and 2-hydroxyethyl radicals may play a role in ethanol-mediated toxicity.

Nonspecific blockade of vascular free radical signals by methylated arginine analogues

Methylated arginine analogues are often used as probes of the effect of nitric oxide; however, their specificity is unclear and seems to be frequently overestimated. This study analyzed the effects of N G -methyl-L-arginine (L-NMMA) on the endothelium-dependent release of vascular superoxide radicals triggered by increased flow. Plasma ascorbyl radical signals measured by direct electron paramagnetic resonance spectroscopy in 25 rabbits increased by 3.8 ñ 0.7 nmol/l vs baseline (28.7 ñ 1.4 nmol/l, P<0.001) in response to papaverine-induced flow increases of 121 ñ 12 por cento. In contrast, after similar papaverine-induced flow increases simultaneously with L-NMMA infusions, ascorbyl levels were not significantly changed compared to baseline. Similar results were obtained in isolated rabbit aortas perfused ex vivo with the spin trap Ó-phenyl-N- tert -butylnitrone (N = 22). However, in both preparations, this complete blockade was not reversed by co-infusion of excess L-arginine and was also obtained by N-methyl-D-arginine, thus indicating that it is not related to nitric oxide synthase. L-arginine alone was ineffective, as previously demonstrated for N G -methyl-L-arginine ester (L-NAME). In vitro , neither L-arginine nor its analogues scavenged superoxide radicals. This nonspecific activity of methylated arginine analogues underscores the need for careful controls in order to assess nitric oxide effects, particularly those related to interactions with active oxygen species