Heme Oxygenase-1 and Autophagy Linked for Cytoprotection.

BACKGROUND: Heme-oxygenase (HO) catalyzes the main enzymatic step of heme degradation and generates anti-inflammatory end products with protective roles in physiological and pathological situations. The importance of HO in pathological conditions is evidenced by its pharmacological inhibition or genetic blockage in different models of stress such as infection, inflammation and oxidative stress. Under these situations, another well-known protective process triggered is autophagy. Autophagy is a homeostatic process that eliminates defective cytosolic components and organelles, allowing cells and tissues to recover through recycling of functional blocks for anabolic reactions. Recently, studies have demonstrated a link between HO activity and autophagy activation. OBJECTIVE: In this review, we focus on the interplay between HO and autophagy, and highlight its importance in homeostasis maintenance under stress conditions.

Nod-like proteins in inflammation and disease.

The field of innate immunity has undergone an enormous upheaval during the last decade. The discovery of different groups of proteins, called pattern recognition molecules (PRMs), which detect microbial components, so-called pathogen-associated molecular patterns (PAMPs) and trigger protective responses, had a huge impact on the understanding of innate immune responses. Among the PRMs, the intracellular Nod-like receptors (NLRs) have recently been identified as key mediators of inflammatory and immune responses. The NLR family is divided into subfamilies on the basis of their different signal transduction domains, and recent studies have highlighted the role of certain NLRs, including Nod1, Nod2, Nalp3, Ipaf and Naip5, in the detection of intracellular microbes and possibly 'danger signals'. In this review, we summarize the current knowledge on the function of these proteins in immunity and inflammation, with a focus on their participation in different disease pathologies.

Phenotypic and genotypic characterisation of Escherichia coli strains serogrouped as enteropathogenic E. coli (EPEC) isolated from pasteurised milk.

Fifty-six Escherichia coli strains, serogrouped as EPEC, isolated from three different brands of pasteurised milk commercialised in Rio de Janeiro, Brazil, were tested for enteropathogenicity markers. Most of the strains (71.4%) were adherent to HEp-2 cells. The adherent strains were distributed among 7 EPEC serogroups (O26, O55, O111, O114, O125, O127, O128, O158). Although almost half of these strains (33.9%) presented unrecognisable adherence phenotypes, classical adherence patterns (localised-like, aggregative and diffuse adherence) described for E. coli and epidemiologically associated with diarrheagenic strains were observed. None of the strains showed typical localised adherence, usually associated with EPEC strains, but 4 of them displayed a localised-like adherence (LAL) phenotype, characterised by fewer and less compact microcolonies but that is still associated with diarrheagenic strains as well as strains of non-human origin. Indeed, 3 of these 4 strains were able to elicit the attaching-effacing lesion (FAS-positive), the central feature of EPEC pathogenesis, and hybridised with bfpA and eae DNA probes. The other LAL-positive strain hybridised with the bfpA probe but gave negative results for the eae probe and FAS assays. Interestingly, all LAL-positive strains produced amplicons of 200 bp in the PCR for bfpA, instead of the expected 326 bp fragment. PCR reactions for stx1 and stx2, two shiga-toxin-encoding genes, gave negative results. Typing of LEE-associated genes by PCR showed the profile eae (beta), tir (beta), espA (alpha) and espB (alpha) for one of the LAL-positive strain. The most prevalent adherence phenotype was the aggregative pattern which is observed in strains epidemiologically associated with persistent diarrhea. Additionally, one strain promoted complete detachment of the Hep-2 cell monolayer after 3 h of infection which might be related to the production of citotoxins, a feature that has been increasingly observed in clinical strains. The possession of EPEC-related O and H antigens is no longer deemed an essential characteristic of true pathogenic EPEC strains, emphasising the importance of routinely screen for virulence markers in E. coli strains isolated from foods. Our results are in accordance with data from the literature that demonstrate that environmental strains display atypical features but yet are capable of eliciting the classical A/E lesion and thus must be considered as potentially pathogenic. Further, our results demonstrate the potential of pasteurised milk as a vehicle for transmission of diarrheagenic E. coli in Brazil.

Cryopreservation of Arachis species by vitrification of in vitro-grown shoot apices and genetic stability of recovered plants.

A storage protocol at cryogenic temperature was established for shoot apices from in vitro plants of the cultivated groundnut (Arachis hypogaea) and wild Arachis species (A. retusa and A. burchellii) using a basic vitrification protocol with direct immersion in liquid nitrogen (LN). The effect of pre-treatments of donor-plants with ABA as well as of different supplements in the post-thaw culture medium was studied. After rapid warming at 40 C, the explants were cultured on MS medium devoid of growth regulators (MS0) or MS supplemented with 4.4(M benzylaminopurine (BAP) and 0.5(M naphthalene acetic acid (NAA) plus 5(M silver nitrate (AgNO3), 0.25% polyvinylpyrrolidone (PVP) or 0.2% activated charcoal. Non-frozen explants from the three species formed one shoot through meristematic amplification when cultured on MS0 medium. These explants also developed callus on MS supplemented with growth regulators (4.4(M BAP and 0.5(M NAA) alone or plus PVP or AgNO3. Callus formation was suppressed in the presence of activated charcoal. Post-thaw regeneration ocurred only through indirect organogenesis on media containing AgNO3 or PVP. Preculturing on medium supplemented with abscisic acid (ABA) improved regrowth rate in these media. Recovery failed to occur in the presence of activated charcoal. The genetic stability of shoots of A. burchellii originated from shoot apices was analyzed through Random Amplified Polymorphic DNA (RAPD) markers.

Effects of androgens on social behavior and morphology of alternative reproductive males of the Azorean rock-pool blenny.

In the Azorean rock-pool blenny Parablennius sanguinolentus parvicornis two sequential reproductive tactics occur. Larger and older males establish breeding territories, while some of the smaller males become attached to nest-holder territories, acting as satellites on these territories, which they help to defend while trying parasitic fertilizations when females go in the nests to spawn. In the present paper we tested the effects of the androgens 17alpha-methyltestosterone (MT) and 11-ketotestosterone (KT) in the expression of male secondary sex characters and bourgeois behavior in satellite males. One week after satellites were implanted with Silastic tubes containing MT, KT, or castor oil (control), androgen-treated satellites had developed male secondary sex traits such as longer and wider male-type genital papilla and anal glands that secrete a sex pheromone, both traits being less expressed or absent, respectively, in satellite males. Androgen treatment had no effect on the gonadosomatic index or on the development of the testicular gland. KT treatment had a positive effect on relative liver weight. In terms of behavior, androgen-implanted individuals were less aggressive both in a mirror test and toward females when these were introduced into their tanks. MT-treated individuals spend more time inside the provided nests. Only androgen-implanted satellites managed to have the females entering their nests. When given a chance in a group tank either to try to attract females to their own nest or to act as satellites of an already established nest-holder's nest, MT-implanted males spent significantly more time in their own nest than near the nest-holder nest. These data suggest that androgens, particularly testosterone, may be involved in mating tactic switching in this species.

11-Ketotestosterone inhibits the alternative mating tactic in sneaker males of the peacock blenny, Salaria pavo.

In the peacock blenny, Salaria pavo, a species with courtship sex-role reversal, smaller, younger males mimic the courtship behavior and the nuptial coloration of females in order to get access to nests during spawning and to parasitize egg fertilization from nest-holder males. Later in their life, sneakers transform both morphologically and behaviorally into nest-holder males. In the present paper we investigate the activational role of 11-ketotestosterone (KT), the most potent androgen in most teleost species, to promote the switch between tactics in sneaker males of S. pavo. Sneakers were implanted either with KT or with control (i.e. castor oil) silastic implants. A week after implantation they were subjected to a set of behavioral tests and morphometric measurements. KT treatment promoted the differentiation of secondary sex characters, such as the anal glands, and inhibited the expression of female courtship behavior. KT-treated sneakers also showed a trend toward less frequent display of female nuptial coloration. There was no effect of KT treatment on the expression of typical nest-holder male behavior. Finally, there was no effect of KT treatment on the number or soma size of arginine vasotocin neurons in the preoptic area, which are often associated with the expression of vertebrate sexual behavior. Thus, KT seems to play a key role in mating tactic switching by inhibiting the expression of female courtship behavior and by promoting the development of male displaying traits (e.g. anal glands). The lack of a KT effect on behavior typical of nest-holding males and vasotocinergic preoptic neurons suggests that a longer time frame or other endocrine/social signals are needed for the initiation of these traits in males that are switching tactics.

Evaluation of enzyme-linked immunosorbent and alternative assays for detection of HIV antibodies using panels of Brazilian sera

Os soros de 472 brasileiros, confirmados como sendo positivos ou negativos em relacao a presenca de anticorpos anti-HIV e compreendendo todo o espectro clinico da infeccao, foram utilizados na avaliaco de seis ensaios imunoenzimaticos comerciais (ELISA), bem como de quatro testes alternativos tais como imunofluorescencia indireta (IFI), hemaglutinacao passiva (HP), dot blot e Karpas AIDS cell test. As sensibilidades variaram de 100 por cento (ELISA Abbott e Roche) a 84,2 por cento (HP) e as especificidades variaram de 99,3 por cento (IFI) a 80,2 por cento (HP). A sensibilidade e especificidade da HP e a sensibilidade do Karpas AIDS cell test foram significativamente menores que os outros ensaios. Embora a IFI e o dot blot tivessem apresentado uma boa sensibilidade e especificidade, os ensaios imunoenzimaticos (ELISA) foram mais adequados para serem utilizados em triagem quando outros parametros tais como facilidade de leitura e interpretacao dos resultados e duracao dos ensaios foram considerados

Evaluation of enzyme-linked immunosorbent and alternative assays for detection of HIV antibodies using panels of Brazilian sera.

Sera from 472 Brazilian subjects, confirmed to be either positive or negative for HIV antibodies and comprising the total clinical spectrum of HIV infection, were utilized in the evaluation of six commercially available enzyme-linked immunosorbent assays (ELISA), as well as of four alternative assays, namely indirect immunofluorescence (IIF), passive hemagglutination (PHA), dot blot and Karpas AIDS cell test. The sensitivities ranged from 100% (Abbott and Roche ELISA) to 84.2% (PHA) and the specificities ranged from 99.3% (IIF) to 80.2% (PHA). The sensitivity and specificity of the PHA and the sensitivity of the Karpas cell test were significantly lower than those of the other tests. Although the IFF and dot blot had good sensitivities and specificities, the six ELISA were more attractive than those tests when other parameters such as ease of reading and duration of assay were considered.

PIP: 6 commercially available ELISA kits and 4 new Brazilian made methods for detecting HIV were compared on 2 panels of sera, 292 from AIDS patients, HIV-positives and negatives, and 180 sera from asymptomatic blood donors, including 90 HIV-positives. The kits tested were 5 ELISAs: Roche Diagnostica (Basel), Hoechst Enzygnostic (Sao Paulo), Virgo Electronuclionics (Columbia MD), Organon Teknika (Boxtel, Netherlands), Salck Industria e Comercio de Produtos Biologicos (Sao Paulo), and a passive hemagglutination test, (Salck Ind), and indirect immunofluorescence IIF (Virgo electronucleonics, Columbia), a dot blot (Embrabio, Empressa Brasiliera de Biotecnologia Ltda, Sao Paolo) and Karpas AIDS cell test, Fujichemical Industries Ltd (Chokeiji, Takaoka, Japan). The sensitivities ranged from 84.2% to 100% with no significant differences in sera from panel A. In panel B, the sensitivity of the PHA test was significantly lower than that of the ELISA and the AIDS cell tests. The specificities of the PHA and the AIDS cell tests were also lower than that of the ELISA. The costs of all the tests were similar, but the equipment needs varied. The simplest tests to perform were the dot blot assay, PHA and Karpas AIDS cell test. The Hoechst ELISA is simpler because it does not require dilution of the serum. The dot takes too long for use in a blood bank, 16-18 hours. Immunofluorescence tests would be practical in countries already screening blood for malaria or Changes disease. Brazil is not doing so on a large scale due to lack of political will. In countries with high incidence of malaria, Chagas disease, leishmania, hepatitis and leprosy, HIV test need to be tested on local sera because of possible B cell activation.