Cryptosporidium and Giardia in urban wastewater: A challenge to overcome.

This study aimed to quantify Giardia and Cryptosporidium in disinfected water reuse samples from two Wastewater Treatment Plants (A and B), which were quantified by USEPA 1693/2014 Method. Giardia was found in 35.8% of the total samples (<0.03 to 16 cysts/L) while Cryptosporidium in 30.2% (<0.03 to 25.8 oocysts/L). This study highlights the presence of both parasites in water for reuse despite treatment processes for their removal, which means there is a challenge to overcome. Their presence is preoccupant even though in low concentrations because the infectivity dose is low coupled with high prevalence in the global population. The practice of water recycling is valuable for sustainable water management and it is in line with Sustainable Developments Goals but should not threaten human health. Tackling this issue is more critical in developing countries because treatment processes are often more limited, the monitoring data from water reuse are not always available, the lack of regulation for water reuse quality and the lack of planning for its sustainable usage.

Impaired decision-making and time perception in individuals with stroke: Behavioral and neural correlates.

Several studies have demonstrated that stroke subjects present impairment of functions related to decision-making and timing, involving the information processing in the neural circuits of the cerebellum in association with the prefrontal cortex. This review is aimed to identify the gaps, and demonstrate a better understanding of decision-making and timing functions in the patients with stroke. Electronic literature database was searched and the findings of relevant studies were used to explore the mechanisms of decision-making and timing in patients with stroke, as well as the circuit connections in timing mediated by prefrontal cortex and cerebellum. A literature review was conducted with 65 studies that synthesized findings on decision-making and time perception in individuals with stroke. Types of neurobiological modalities in this study included: Relationships among decision-making, time perception, related cognitive aspects (such as discrimination tasks, verbal estimation, bisection tasks, time production and motor reproduction), and motor control. We demonstrate that the timing processes are important for the performance in cognitive tasks and that the cerebellum and prefrontal cortex are involved in decision-making and time perception. In the context, the decision-making is impaired in stroke patients has a great impact on executive functions, and this seems to be important in determining neurobiological aspects relevant to the time interval interpretation.

Glutathione peroxidase and glutathione S-transferase in blood and liver from a hypoxia-tolerant fish under oxygen deprivation.

Liver enzyme activities can be employed as biomarkers, but liver can only be obtained with death of the specimen. On the other hand, blood withdrawal is a non-lethal procedure. Accordingly, the hypothesis of this study is to verify if glutathione peroxidase (GPX) and glutathione S-transferase (GST) activities in blood parallel those in the liver of the hypoxia-tolerant fish, Piaractus mesopotamicus (pacu), submitted to hypoxia conditions. GPX was assayed with H2O2 in cytosols from both liver and erythrocytes and exhibited no significant variation, either in erythrocytes or in liver, when comparing pacus under normoxia with those under hypoxia (42 h). GST activity with chloro-dinitrobenzene (CDNB), an artificial substrate suitable for almost all GST isoenzymes, was compared to activity with 4-hydroxy-nonenal (4-HNE), a physiological endogenous substrate. GST activity with CDNB did not change in liver or in erythrocyte cytosols in pacus under hypoxia compared to those under normoxia. On the other hand, a significant decrease in erythrocyte activity with 4-HNE was observed after 42 h of hypoxia in both erythrocytes and liver, which may be a response to increased lipid oxidation in erythrocytes. Erythrocyte GST activity was 3-fold higher with 4-HNE than with CDNB, indicating that 4-HNE is a more appropriate substrate to determine GST activity in pacu erythrocytes.

Genotoxicity of citrate-coated silver nanoparticles to human keratinocytes assessed by the comet assay and cytokinesis blocked micronucleus assay.

Silver nanoparticles (AgNPs) are widely used in industrial, cosmetic, and biomedical products, and humans are frequently exposed to these products through the skin. It is widely recognized that the characteristics of AgNPs (e.g., size, coating) may influence their cytotoxic effects, but their correlation with DNA damage and mitotic disorders remains poorly explored. In this study, human keratinocytes (HaCaT cell line) were exposed to well-characterized 30 nm AgNPs coated with citrate, and their effects on viability, DNA fragmentation (assessed by the comet assay), and micronuclei (MNi) induction (assessed by the cytokinesis-block micronucleus cytome assays, CBMN) were investigated. The results showed that 10 and 40 µg/mL AgNPs decreased cell proliferation and viability, and induced a significant genetic damage. This was observed by an increase of DNA amount in comet tail, which linearly correlated with dose and time of exposure. Also, cytostaticity (increase of mononucleated cells) and MNi rates increased in treated cells. In contrast, no significant changes were observed in nucleoplasmatic bridges (NPBs) or nuclear buds (NBUDs), although NBUDs tended to increase in all conditions and periods. The cytostatic effects on HaCaT cells were also shown by the decrease of their nuclear division index. Thus, both comet and CBMN assays supported the observation that citrate-AgNPs induced genotoxic effects on HaCaT cells. Considering that AgNPs are present in a vast number of consumer products and also in multiple nanomedicine skin applications and formulations, more research is needed to determine the properties that confer less toxicity of AgNPs to different cell lines.

The influence of Citrate or PEG coating on silver nanoparticle toxicity to a human keratinocyte cell line.

Surface coating of silver nanoparticles may influence their toxicity, in a way yet to decipher. In this study, human keratinocytes (HaCaT cells) were exposed for 24 and 48h to well-characterized 30nm AgNPs coated either with citrate (Cit30 AgNPs) or with poly(ethylene glycol) (PEG30 AgNPs), and assessed for cell viability, reactive oxygen species (ROS), cytokine release, apoptosis and cell cycle dynamics. The results showed that Cit30 AgNPs and PEG30 AgNPs decreased cell proliferation and viability, the former being more cytotoxic. The coating molecules per se were not cytotoxic. Moreover, Ag(+) release and ROS production were similar for both AgNP types. Cit30 AgNPs clearly induced apoptotic death, while cells exposed to PEG30 AgNPs appeared to be at an earlier phase of apoptosis, supported by changes in BAX, BCL2 and CASP-3 expressions. Concerning the impact on cell cycle dynamics, both Cit30 and PEG30 AgNPs affected cell cycle regulation of HaCaT cells, but, again, citrate-coating induced more drastic effects, showing earlier downregulation of cyclin B1 gene and cellular arrest at the G2 phase. Overall, this study has shown that the surface coating of AgNPs influences their toxicity by differently regulating cell-cycle and cell death mechanisms.

Melatonin affects conjugation of 4-hydroxynonenal with glutathione in liver of pacu, a hypoxia-tolerant fish.

In cytosol from liver of pacu, Piaractus mesopotamicus, a hypoxia-tolerant fish that dwells in Pantanal, we found an enzyme activity capable of modulating the alkenal 4-hydroxy-2-nonenal (HNE) by conjugating it with glutathione (GST-HNE activity). HNE is a downstream metabolite from the oxidation of polyunsaturated fatty acids by reactive oxygen species arisen from mitochondria of animal cells. HNE production may increase more intensively under oxidative stress. Harmful effects to cell survival may occur when HNE increases over 10(-4) M. Pacus submitted to hypoxia in July (cold season in Pantanal) showed 40% less of this GST-HNE conjugating activity in their liver cytosol. Injecting pacus subjected to hypoxia during the cold season with a summer physiological dose of melatonin caused their liver cytosolic GST-HNE activity to increase up to the levels found in the warm season. From October to March (warm season in Pantanal), pacus are prone to oxidative stress particularly during potamodromous active oxygen-demanding swimming, when they migrate up rivers to spawn. Thus, our findings point out that the higher levels of melatonin in circulation during the summer are important to avoid the increase of 4-HNE inside liver cells of this fish species.

Enzymatic GST levels and overall health of mullets from contaminated Brazilian Lagoons.

Glutathione S-transferase (GST) assays in non-mammalian organisms are usually conducted inappropriately, since no previous standardization of the optimal concentrations of proteins and substrates and adequate pH is conducted. Standardization is a key task to adjust enzyme assays at their kinetically correct maximal initial velocities, if one wants these velocities to indicate the amount of enzyme in a sample. In this paper GST assays were standardized in liver cytosol to compare seasonal GST levels in liver of mullet from two contaminated lagoons in the Rio de Janeiro to those from a reference bay. GST potential as a biomarker of sublethal intoxication in this species was also evaluated. Mullet liver GST levels assayed with substrates that corresponded to three different GST isoenzymes varied throughout the year. The differences indicated that mullets are suffering from sublethal intoxication from contaminants in these lagoons. Seasonal variations of activity were relevant, since these could indicate differences in xenobiotic input into the areas. An analysis of overall mullet health condition using a morphological index (the Fulton Condition Factor) and macroscopic abnormalities corroborated the differences in GST levels, with fish from one of the sites in worse overall health condition showing lower and significantly different FCF when compared to the reference site. Therefore, GST standardized activity levels are useful biomarkers of environmental contamination for mullet.

Talking about bioelectrical potentials using rings of the mesenteric artery without glass micropipettes.

In the present study, a practical activity is proposed to adopt an experimental approach to demonstrate the relationship between the equilibrium potential for K(+) and transmembrane electrical potential without glass micropipettes. A conventional setup for recording contractile activity of isolated smooth muscle preparations was used based on the events elegantly described by Somlyo and Somlyo in the 1960s. They showed that, in response to a given stimulus, smooth muscle cells may contract, recruiting electromechanical or pharmacomechanical coupling by mechanisms that involve, or not, changes in transmembrane potential, respectively. By means of contractions and relaxations of a ring-like preparation from the rat mesenteric artery, it is possible to observe the functional consequences of handling K(+) concentration in the extracellular compartment and the effects caused by opening K(+) channels in that preparation, which are significant when the cell membrane establishes an electrical potential difference between intra- and extracellular compartments (driven mainly by K(+) permeability under resting conditions). The effects observed by students fit well with values predicted by Nernst and Goldman-Hodgin-Katz equations, and we demonstrated that the activity is able to improve students' comprehension regarding basic principles of bioelectricity.

In vitro and in vivo toxicological evaluation of extract and fractions from Baccharis trimera with anti-inflammatory activity.

ETHNOPHARMACOLOGICAL RELEVANCE: Baccharis trimera (Less) DC. (Asteraceae), popularly known in Brazil as "carqueja", have been used in folk medicine to treat gastrointestinal, hepatic and renal diseases, and inflammatory processes as rheumatism. AIM OF THE STUDY: To evaluate the in vitro and in vivo toxicological effects of anti-inflammatory Baccharis trimera aqueous extract and fractions. MATERIALS AND METHODS: Aqueous extract of Baccharis trimera (AEBt) was produced by infusion in boiling water. After lyophylization AEBt was extracted with 80% ethanol, originating the ethanolic supernatant fraction (EFBt) and the aqueous sediment fraction (AFBt). Anti-inflammatory properties of AEBt, EFBt or AFBt (3, 30 or 300 µg/kg b.w.) were evaluated by the carrageenan-induced mouse paw edema using indomethacin (10mg/kg) as positive control. The growth of rat hepatoma cells (HTC) and human embryo kidney epithelial cells (HEK) was determined by protein staining assay. Cytotoxicity was assayed by the tetrazolium salt (MTT) reduction. Cyclosporin was used as reference cytotoxic drug for spleen cells and doxorubicin for HTC and HEK cells. For in vivo toxicological evaluation SW male mice were daily and oral (gavage) treated with extract/fractions at 4.2mg/kg or 42 mg/kg during 15 days. After treatment liver or kidney cells were submitted to comet assay to determine the DNA damage index, and the glutathione S-transferase activity was assayed towards ETHA (class Pi) and CDNB (several classes). Mutagenicity was evaluated by the Ames test using Salmonella typhimurium strains TA97, TA98, TA100, and TA102. RESULTS: The anti-inflammatory effects of EFBt were higher than those of AEBt or AFBt. Mice treatment (3-300 µg/kg) with AFBt reduced the paw edema (3h) at lower levels (29.2-37.3%; P<0.01), than those observed for AEBt (44.7-54.2%; P<0.001), EFBt (49.3-58.2%; P<0.001) or indomethacin (64.6%, P<0.001, 10mg/kg). The growth of kidney cells (HEK) was inhibited by AEBt (IC(50) 182.6 µg/ml), EFBt (IC(50) 78.1 µg/ml) and AFBt (IC(50) 86.2 µg/ml), with lower effects on HTC hepatic cell (IC(50) 308.8 µg/ml, 396.5 µg/ml and 167.9 µg/ml, respectively). As evaluated by MTT test, AFBt exhibited cytotoxicity for HEK cells (IC(50) 372.5 µg/ml), but none for HTC ones; by the way, AFBt stimulated spleen cells (EC(50) 2.2 µg/ml) while cyclosporine, a cytotoxic reference drug inhibited them with IC(50) of 0.42 µg/ml; the IC(50) for doxorubicin for HEK and HTC cells was 0.28 µg/ml and 14.4 µg/ml, respectively, at 96h. No mutagenic potential was observed. Mice treatment with AEBt or AFBt at 42 mg/kg for 15 days altered the kidney relative weight, but not at 4.2mg/kg. Baccharis trimera did not change liver, spleen or popliteal lymph node relative weight. DNA damage index of kidney cells was observed on mice treated with AEBt/AFBt, but not on animals treated with EFBt, while DNA lesions were detected on liver cells only after AFBt treatment. The general activities of hepatic GST and Pi GST were reduced by EFBt and AFBt treatment, respectively. CONCLUSIONS: Baccharis trimera did not show mutagenicity, inhibited the GST activity, a hepatic detoxification enzyme, and induced in vivo (genotoxicity) and in vitro toxicological effects to kidney cells.

Dispensação farmacêutica: uma análise dos conceitos legais em relação à prática profissional / Pharmaceutical dispensing: an analysis of legal concepts in relation to professional practice

O âmbito profissional do farmacêutico sofreu mudanças ao longo dos anos, provocando a descaracterização das farmácias comunitárias em virtude da acentuada concepção mercantil que vem apresentando e do distanciamento do farmacêutico na relação direta com a comunidade. Dessa forma, a dispensação de medicamentos se tornou um ato mecânico, desprovido dos cuidados necessários para a assistência à saúde, havendo, contudo, uma tendência de incorporação de novas práticas. Nesse sentido, ao estudar os conceitos de dispensação explicitados nas legislações sanitárias e profissionais brasileiras, esse artigo tem como objetivo analisá-los em relação ao contexto atual. Trata-se de um estudo teórico, realizado com a técnica da análise de conteúdo. Os resultados mostram que apesar das necessidades e novas demandas apontadas no cenário farmacêutico, ocorreram poucas mudanças nos conceitos de dispensação nas legislações estudadas, apontando a necessidade de se reafirmar a dispensação como atividade inserida no contexto da assistência à saúde.

The pharmacist?s professional scope of practice has changed over the years, causing a loss of the special role of community pharmacies, due to the strongly market-oriented conception that is emerging and the distancing of the pharmacist from a direct relationship with the community. Thus, the dispensing of drugs has become a mechanical act, devoid of the care necessary for health provision, there being, however, a tendency to incorporate new practices. Therefore, by studying the concepts of dispensing spelled out in the health laws and by health professionals in Brazil, the aim of this article is to analyze them in relation to the current context. This is a theoretical study, carried out by the technique of content analysis. The results show that despite the new demands and requirements noted in the pharmaceutical scenario, there have been few changes in the concepts of dispensing in the laws examined, indicating a need to reaffirm dispensing as an activity that has a place within the context of health care.

[The effects of bromazepam on the performance of a sensory-motor activity: an electroencephalographic study]. / Efectos del bromacepam en el desarrollo de una actividad sensoriomotora: un estudio electroencefalográfico.

AIMS: To investigate the effects of using bromazepam on the relative power in alpha while performing a typing task. Bearing in mind the particularities of each brain hemisphere, our hypothesis was that measuring the relative power would allow us to investigate the effects of bromazepam on specific areas of the cortex. More specifically, we expected to observe different patterns of powers in sensory-motor integration, attention and activation processes. SUBJECTS AND METHODS: The sample was made up of 39 subjects (15 males and 24 females) with a mean age of 30 +/- 10 years. The control (placebo) and experimental (3 mg and 6 mg of bromazepam) groups were trained in the typing task with a randomised double-blind model. RESULTS: A three-way ANOVA and Scheffé test were used to analyse interactions between the factors condition and moment, and between condition and sector. CONCLUSIONS: The doses used in this study facilitated motor performance of the typing task. In this study, the use of the drug did not prevent learning of the task, but it did appear to concentrate mental effort on more restricted and specific aspects of typing. It also seemed to influence the rhythm and effectiveness of the operations performed during mechanisms related to the encoding and storage of new information. Likewise, a predominance of activity was observed in the left (dominant) frontal area in the 3 mg bromazepam group, which indicates that this dose of the drug affords the subject a greater degree of directionality of cortical activity for planning and performing the task.

Efectos del bromacepam en el desarrollo de una actividad sensoriomotora: un estudio electroencefalográfico / The effects of bromazepan on the performance of a sensory-motor activity: an electroencephalographic study

Objetivos. Investigar los efectos del uso del bromacepam en la potencia relativa en alfa durante la realización de unatarea de mecanografía. Teniendo en cuenta las particularidades de cada hemisferio cerebral, nuestra hipótesis era que a travésde la medida de la potencia relativa sería posible investigar el efecto del bromacepam sobre áreas corticales específicas. Concretamente,se esperaba observar diferentes patrones de potencias en los procesos de atención, activación e integración sensoriomotora.Sujetos y métodos. La muestra estaba formada por 39 sujetos (15 hombres y 24 mujeres) con una media de edad de30 ± 10 años. Los grupos control (placebo) y experimental (bromacepam de 3 mg y 6 mg) fueron entrenados en la tarea de mecanografíacon un modelo doble ciego aleatorizado. Resultados. Mediante el ANOVA de tres vías y el test de Scheffé se comprobaroninteracciones entre los factores condición y momento y entre condición y sector. Conclusión. Las dosis empleadas en esteestudio facilitaron el desarrollo motor de la tarea de mecanografía. En este estudio, el uso del fármaco no impidió el aprendizajede la tarea, pero parece ser que concentró el esfuerzo mental sobre aspectos más restringidos y específicos de la mecanografía.Tuvo lugar una influencia sobre el ritmo y la eficacia de las operaciones ejecutadas durante mecanismos de codificacióny almacenamiento de nuevas informaciones. Asimismo, se comprobó un predominio de actividad en el área frontal izquierda(dominante) en el grupo bromacepam 3 mg, lo cual indica que esta dosis del fármaco permite al sujeto un mayor direccionamientode la actividad cortical para la planificación y la ejecución de la tarea(AU)

Aims. To investigate the effects of using bromazepam on the relative power in alpha while performing a typing task.Bearing in mind the particularities of each brain hemisphere, our hypothesis was that measuring the relative power wouldallow us to investigate the effects of bromazepam on specific areas of the cortex. More specifically, we expected to observedifferent patterns of powers in sensory-motor integration, attention and activation processes. Subjects and methods. The samplewas made up of 39 subjects (15 males and 24 females) with a mean age of 30 ± 10 years. The control (placebo) and experimental(3 mg and 6 mg of bromazepam) groups were trained in the typing task with a randomised double-blind model. Results. A threewayANOVA and Scheffé test were used to analyse interactions between the factors condition and moment, and betweencondition and sector. Conclusions. The doses used in this study facilitated motor performance of the typing task. In this study,the use of the drug did not prevent learning of the task, but it did appear to concentrate mental effort on more restricted andspecific aspects of typing. It also seemed to influence the rhythm and effectiveness of the operations performed duringmechanisms related to the encoding and storage of new information. Likewise, a predominance of activity was observed in theleft (dominant) frontal area in the 3 mg bromazepam group, which indicates that this dose of the drug affords the subject agreater degree of directionality of cortical activity for planning and performing the task(AU)

[The role played by the parietooccipital cortex in the process of sensory-motor integration: an electroencephalographic study]. / Participación de la corteza parietooccipital en el proceso de integración sensoriomotora: estudio electroencefalográfico.

INTRODUCTION: A fundamental aspect of planning future actions is the performance and control of motor tasks. This behaviour is done through sensory-motor integration. AIM: To explain the electrophysiological mechanisms in the cortex (modifications to the alpha band) that are involved in anticipatory actions when individuals have to catch a free-falling object. SUBJECTS AND METHODS: The sample was made up of 20 healthy subjects of both sexes (11 males and 9 females) with ages ranging between 25 and 40 years (32.5 +/- 7.5) who were free of mental or physical diseases (previous medical history); the subjects were right-handed (Edinburgh Inventory) and were not taking any psychoactive or psychotropic substances at the time of the study. The experiment consisted in a task in which subjects had to catch freely falling objects. The experiment was made up of six blocks of 15 tests, each of which lasted 2 minutes and 30 seconds, with a break of one minute between blocks. Data were captured by means of a quantitative electroencephalogram two seconds before and two seconds after each ball was dropped. RESULTS: An interaction of the factors moment and position was only observed for the right parietooccipital cortex, in the combination of electrodes P4-O2. CONCLUSION: These findings suggest that the right parietooccipital cortex plays an important role in increasing expectation and swiftness in the processes of preparing for a motor task.

[Learning and implicit memory: mechanisms and neuroplasticity]. / Aprendizaje y memoria implícita: mecanismos y neuroplasticidad.

INTRODUCTION: Learning and memory are complex processes that researchers have been attempting to unravel for over a century in order to gain a clear view of the underlying mechanisms. AIMS: To review the basic cellular and molecular mechanisms involved in the process of procedural retention, to offer an overall view of the fundamental mechanisms involved in storing information by means of theories and models of memory, and to discuss the different types of memory and the role played by the cerebellum as a modulator of procedural memory. DEVELOPMENT: Experimental results from recent decades have opened up new areas of study regarding the participation of the biochemical and cellular processes related to the consolidation of information in the nervous system. CONCLUSIONS: The neuronal circuits involved in acquiring and consolidating memory are still not fully understood and the exact location of memory in the nervous system remains unknown. A number of intrinsic and extrinsic factors interfere in these processes, such as molecular (long-term potentiation and depression) and cellular mechanisms, which respond to communication and transmission between nerve cells. There are also factors that have their origin in the outside environment, which use the association of events to bring about the formation of new memories or may divert the subject from his or her main focus. Memory is not a singular occurrence; it is sub-divided into declarative and non-declarative or, when talking about the time it lasts, into short and long-term memory. Moreover, given its relation with neuronal mechanisms of learning, memory cannot be said to constitute an isolated process.

Inhibition of HSV-1 replication and HSV DNA polymerase by the chloroxoquinolinic ribonucleoside 6-chloro-1,4-dihydro-4-oxo-1-(beta-D-ribofuranosyl) quinoline-3-carboxylic acid and its aglycone.

We describe in this paper that the synthetic chloroxoquinolinic ribonucleoside 6-chloro-1,4-dihydro-4-oxo-1-(beta-D-ribofuranosyl) quinoline-3-carboxylic acid (compound A) and its free aglycogene base (compound B) inhibit, with low cytotoxicity, the replication of herpes simplex virus type 1 and 2 (HSV-1 and HSV-2). Compound A inhibited HSV-1 replication in Vero cells with an EC(50) of 1.3 and 1.4 microM for an acyclovir (ACV)-sensitive strain and an ACV-resistant strain of this virus, respectively. Additionally, it inhibited HSV-2 replication with an EC(50) of 1.1 microM. Compound B also inhibited the ACV-sensitive and -resistant HSV-1 strains, and HSV-2 at EC(50) values of 1.7, 1.9 and 1.6 microM, respectively. Time-of-addition assays, performed with compound A, suggested that this molecule at an early time point of the HSV replication cycle. Kinetic assays demonstrated that compounds A and B inhibit the HSV DNA polymerase activity in a noncompetitive fashion, with a K(i) equal to 0.1 and 0.2 microM, respectively. Taken together, our results suggest that compounds A and B represent promising lead molecules for further anti-HSV drug design.

Cytosolic glutathione peroxidase from liver of pacu (Piaractus mesopotamicus), a hypoxia-tolerant fish of the Pantanal.

Pacu (Piaractus mesopotamicus Holmberg, 1887, Characiformes) dwells in waters of Pantanal, in which it has adapted for alternate concentrations of dissolved oxygen. Intracellular antioxidant protection should be vital for such an adaptation. Accordingly, we found that cytosol from liver of pacu has the highest antioxidant glutathione peroxidase activity so far reported for fish and murine species. To clarify whether this activity was due to a selenium independent glutathione S-transferase or to a glutathione peroxidase, we purified it and studied its kinetics. The substrates cumene hydroperoxide and hydrogen peroxide were promptly reduced by the enzyme, but peroxidized phosphatidylcholine had to undergo previous fatty acid removal with phospholipase A(2). Augmenting concentrations (from 2 to 6 mM) of reduced glutathione activated the pure enzyme. Curves of velocity versus different micromolar concentrations of hydrogen peroxide in the presence of 2, 4 or 8 mM reduced glutathione indicated that at least 2.5 mM reduced glutathione should be available in vivo for an efficient continuous destruction of micromolar concentrations of hydrogen peroxide by this peroxidase. Molecular exclusion HPLC and SDS-polyacrylamide gel electrophoresis indicated that the purified peroxidase is a homotetramer. Data from internal sequences showed selenocysteine in its primary structure and that the enzyme was a homologue of the type-1 glutathione peroxidase found in rat, bull, trout, flounder and zebra fish. Altogether, our data establish that in liver cells of pacu, a hypoxia-tolerant fish from South America, there are high levels of a cytosolic GPX-1 capable of quenching hydrogen peroxide and fatty acid peroxides, providing an effective antioxidant action.

The chloroxoquinolinic derivative 6-chloro-1,4-dihydro-4-oxo-1-(beta-D-ribofuranosyl) quinoline-3-carboxylic acid inhibits HSV-1 adsorption by impairing its adsorption on HVEM.

In this paper, we describe that the oxoquinolinic acid derivative (compound A) inhibited HSV-1 adsorption on Vero cells. This effect was achieved with an EC(50) value of 10 +/- 2.0 microM and with low cytotoxicity, since the CC(50) value for compound A was >1000 microM. Moreover, we demonstrate for the first time that adsorption inhibition was due to the blockage of the interactions between HSV-1 and the cellular receptor herpes virus entry mediator (HVEM). These results show that compound A can prevent HSV-1 infection in Vero cells, encouraging further studies to determine at what level compound A inhibits HSV-1-HVEM interactions.

A novel butyrylcholinesterase from serum of Leporinus macrocephalus, a Neotropical fish.

We show here that serum of piaussu, a Neotropical characin fish, has the highest butyrylcholinesterase activity so far described for humans and fish. To clarify whether this cholinesterase could protect piaussu against anticholinesterase pesticides by scavenging organophosphates, we purified it 1700-fold, with a yield of 80%. Augmenting concentrations (from 0.01 to 20 mM) of butyrylthiocholine activated it. The pure enzyme was highly inhibited by chlorpyriphos-oxon (ki=10,434x10(6) M-1 min-1) and by the specific butyrylcholinesterase inhibitor, isoOMPA (ki=45.7x10(6) M-1 min-1). Electrophoresis of total serum and 2-D electrophoresis of the purified cholinesterase showed that some enzyme molecules could circulate in piaussu serum as heterogeneously glycosylated dimers. The enzyme's N-terminal sequence was similar to sequences found for butyrylcholinesterase from sera of other vertebrates. Altogether, our data present a novel butyrylcholinesterase with the potential of protecting a fish from poisoning by organophosphates.