Continuous reproduction and recruitment in the narrowback mud crab Panopeus americanus (Brachyura, Panopeidae) in a remnant human-impacted mangrove area

The population structure, reproductive period, and juvenile recruitment of Panopeus americanus were studied in order to enhance knowledge of its life cycle and reproductive strategy and promote the maintenance of its natural stocks in an impacted region. Specimens were collected in the remnant human-impacted mangrove at Araca, state of Sao Paulo, Brazil, at two-month intervals from January to November 2005, at low tide, with a capture effort by three people. The crabs were measured (carapace width, CW) and sexed. The total catch was 275 animals, including 132 males (48.0%); 127 females (46.2%), of which 39 were ovigerous (14.2% of total catch); and 16 individuals whose sex could not be identified (5.8%). No correlation was observed between water temperature and the number of collected individuals; however, there was a significant, positive correlation with salinity. Males were significantly larger than females. The size-frequency distribution was bimodal, reflecting the occurrende of more than one recruitment pulse and the differential abundance of adults during the period of study. The overall sex ratio was 1:0.97 favoring males, and was not significantly different from the expected value, i.e., this population fits the anomalous pattern of sex occurrence in size classes. Ovigerous females were captured in all sampled months, which explain the continuous recruitment observed. Expected low levels of individuals of different size classes in the population were not confirmed. All population aspects found here allowed us to infer that this population of P. americanus is well established in the impacted mangrove by virtue of its reproductive strategy.

Anticlastogenic and antigenotoxic effects of selenomethionine on doxorubicin-induced damage in vitro in human lymphocytes

The use of antioxidants during chemotherapy has been shown to reduce or prevent the undesirable effects experienced by healthy cells. Micronutrient selenium is well known for its antioxidant properties; however, selenium exhibits a bimodal nature in that both its beneficial and toxic properties lie within a limited and narrow dose range. The present study investigated the possible protective effects of selenomethionine (SM) on the cytotoxicity, genotoxicity and clastogenicity of the chemotherapic doxorubicin (DXR), a key chemotherapic used in cancer treatment. Human peripheral lymphocytes were treated in vitro with varying concentrations of SM (0.25 mu M, 0.5 mu M, 1.0 mu M and 2.0 mu M), tested in combination with DXR (0.15 mu g/mL). SM alone was not cytotoxic and when combined with DXR treatment, reduced the DNA damage index significantly, the frequency of chromosomal aberrations, the number of aberrant metaphases and the frequency of apoptotic cells. The mechanism of chemoprotection of SM may be related to its antioxidant properties as well as its ability to interfere with DNA repair pathways. Therefore this study showed that SM is effective in reducing the genetic damage induced by the antitumoral agent DXR. (C) 2007 Elsevier Ltd. All rights reserved.

Mandibular gland secretions of meliponine worker bees: further evidence for their role in interspecific and intraspecific defence and aggression and against their role in food source signalling

Like ants and termites some species of stingless bees (Meliponini), which are very important pollinators in the tropics, use pheromone trails to communicate the location of a food source. We present data on the communicative role of mandibular gland secretions of Meliponini that resolve a recent controversy about their importance in the laying of such trails. Volatile constituents of the mandibular glands have been erroneously thought both to elicit aggressive/defensive behaviour and to signal food source location. We studied Trigona spinipes and Scaptotrigona aff. depilis (`postica`), two sympatric species to which this hypothesis was applied. Using extracts of carefully dissected glands instead of crude cephalic extracts we analysed the substances contained in the mandibular glands of worker bees. Major components of the extracts were 2-heptanol (both species), nonanal (T. spinipes), benzaldehyde and 2-tridecanone (S. aff. depilis). The effect of mandibular gland extracts and of individual components thereof on the behaviour of worker bees near their nest and at highly profitable food sources was consistent. Independent of the amount of mandibular gland extract applied, the bees overwhelmingly reacted with defensive behaviour and were never attracted to feeders scented with mandibular gland extract or any of the synthetic chemicals tested. Both bee species are capable of using mandibular gland secretions for intra-and interspecific communication of defence and aggression and share 2-heptanol as a major pheromone compound. While confirming the role of the mandibular glands in nest defence, our experiments provide strong evidence against their role in food source signalling.

Disposition of MDMA and Metabolites in Human Sweat Following Controlled MDMA Administration

BACKGROUND: Understanding the excretion of 3,4-methylenedioxymethamphetamine (MDMA) and metabolites in sweat is vital for interpretation of sweat tests in drug treatment, criminal justice, and workplace programs. METHODS: Placebo, low (1.0 mg/kg), and high (1.6 mg/kg) doses of oral MDMA were given double-blind in random order to healthy volunteers (n = 15) with histories of MDMA use. Participants resided on the closed clinical research unit for up to 7 days after each dose. Volunteers wore PharmChek (R) sweat patches (n = 640) before, during, and after controlled dosing. Patches were analyzed by solid phase extraction and GC-MS for MDMA, methylenedioxyamphetamine (MDA), 4-hydroxy-3-methoxyamphetamine (HMA), and 4hydroxy-3-methoxymethamphetamine (HMMA). Limits of quantification (LOQ) were 2.5 ng/patch for MDMA and 5 ng/patch for HMA, HMMA, and MDA. RESULTS: MDMA was the primary analyte detected in 382 patches (59.7%), with concentrations up to 3007 ng/patch. MDA was detected in 188 patches (29.4%) at <172 ng/patch, whereas no HMMA or HMA was detected; 224 patches (35.0%) and 60 patches (9.4%) were positive for MDMA and MDA, respectively, at the 25-ng/patch threshold proposed by the Substance Abuse and Mental Health Services Administration. CONCLUSIONS: Sweat testing was shown to be an effective and reliable method for monitoring MDMA use in this controlled MDMA administration study. However, variability in sweat excretion suggests that results should be interpreted qualitatively rather than quantitatively. These data provide a scientific database for interpretation of MDMA sweat test results. (C) 2008 American Association for Clinical Chemistry

In vitro photodynamic therapy on human oral keratinocytes using chloroaluminum-phthalocyanine

In this study, oral carcinoma cells were used to evaluate chloroaluminum-phthalocyanine encapsulated in liposomes as the photosensitizer agent in support of photodynamic therapy (PDT). The genotoxicity and cytotoxicity behavior of the encapsulated photosensitizer in both dark and under irradiation using the 670-nm laser were investigated with the classical trypan blue cell viability test, the acridine orange/ethidium bromide staining organelles test, micronucleus formation frequency, DNA fragmentation, and cell morphology. The cell morphology investigation was carried out using light and electronic microscopes. Our findings after PDT include reduction in cell viability (95%) associated with morphologic alterations. The neoplastic cell destruction was predominantly started by a necrotic process, according to the assay with acridine orange and ethidium bromide, and this was confirmed by electronic microscopy analysis. Neither the PDT agent nor laser irradiation alone showed cytotoxicity, genotoxicity, or even morphologic alterations. Our results reinforce the efficiency of tight-irradiated chloroaluminum-phthalocyanine in inducing a positive effect of PDT. (C) 2008 Elsevier Ltd. All rights reserved.

In-tube solid-phase microextraction coupled to liquid chromatography (in-tube SPME/LC) analysis of nontricyclic antidepressants in human plasma

A sensitive, selective, and reproducible in-tube solid-phase microextraction and liquid chromatographic (in-tube SPME/LC-UV) method for simultaneous determination of mirrazapine, citalopram, paroxetine, duloxetine, fluoxetine, and sertraline in human plasma was developed, validated and further applied to the analysis of plasma samples from elderly patients undergoing therapy with antidepressants. Important factors in the optimization of in-tube SPME efficiency are discussed, including the sample draw/eject volume, draw/eject cycle number, draw/eject flow-rate, sample pH, and influence of plasma proteins. The quantification limits of the in-tube SPME/LC method varied between 20 and 50 ng/mL, with a coefficient of variation lower than 10%. The response of the in-tube SPME/LC method for most of the drugs was linear over a dynamic range from 50 to 500 ng/mL, with correlation coefficients higher than 0.9985. The in-tube SPME/LC can be successfully used to analyze plasma samples from ageing patients undergoing therapy with nontricyclic antidepressants. (c) 2007 Elsevier B.V. All rights reserved.

Microextraction in packed sorbent for analysis of antidepressants in human plasma by liquid chromatography and spectrophotometric detection

A sensitive and reproducible method by microextraction packed sorbent and liquid chromatography with UV detection (MEPS/LC-UV) is described for the determination of new generation antidepressants (sertraline, mirtazapine, fluoxetine, citalopram and paroxetine) in human plasma samples. The MEPS variables, such as sample volume, pH, number of extraction cycles (draw-eject), and desorption conditions (solvent and solvent volume of elution) influenced the MEPS/LC efficiency significantly. Important factors in the optimization of MEPS efficiency, as well as washing steps and carryover effect are discussed. The analyses were carried out using small sample volumes (400 mu L.), and in a short time period (3 min for the entire sample preparation step). The MEPS/LC-UV method was shown to be linear at concentrations ranging from the limit of quantification (LOQ) to 1000 ng mL(-1). The LOQ values ranged from 10 to 25 ng mL(-1). The inter-day precision of the method presented coefficient of the variation ranging from 1.3% to 8.7%. On the basis of analytical validation, it is shown that the MEPS/LC-UV methodology is adequate for antidepressant analysis, from therapeutic to toxic levels. In order to evaluate the proposed method for clinical use, the MEPS/LC-UV method was applied to analysis of plasma samples from elderly depressed patients. (C) 2010 Elsevier B.V. All rights reserved.

Determination of fluoxetine and norfluoxetine enantiomers in human plasma by polypyrrole-coated capillary in-tube solid-phase microextraction coupled with liquid chromatography-fluorescence detection

A sensitive, selective, and reproducible in-tube polypyrrole-coated capillary (PPY) solid-phase microextraction and liquid chromatographic method for fluoxetine and norfluoxetine enantiomers analysis in plasma samples has been developed, validated, and further applied to the analysis of plasma samples from elderly patients undergoing therapy with antidepressants. Important factors in the optimization of in-tube SPME efficiency are discussed, including the sample draw/eject volume, draw/eject cycle number, draw/eject flow-rate, sample pH, and influence of plasma proteins. Separation of the analytes was achieved with a Chiralcel OD-R column and a mobile phase consisting of potassium hexafluorophosphate 7.5 mM and sodium phosphate 0.25 M solution, pH 3.0, and acetonitrile (75:25, v/v) in the isocratic mode, at a flow rate of 1.0 mL/min. Detection was carried out by fluorescence absorbance at Ex/Em 230/290 nm. The multifunctional porous surface structure of the PPY-coated film provided high precision and accuracy for enantiomers. Compared with other commercial capillaries, PPY-coated capillary showed better extraction efficiency for all the analytes. The quantification limits of the proposed method were 10 ng/mL for R- and S-fluoxetine, and 15 ng/mL for R- and S-norfluoxetine, with a coefficient of variation lower than 13%. The response of the method for enantiomers is linear over a dynamic range, from the limit of quantification to 700ng/mL, with correlation coefficients higher than 0.9940. The in-tube SPME/LC method can therefore be successfully used to analyze plasma samples from ageing patients undergoing therapy with fluoxetine. (C) 2009 Elsevier B.V. All rights reserved.

Refolding and purification of the human secreted group IID phospholipase A(2) expressed as inclusion bodies in Escherichia coli

The secreted phospholipases A(2) (sPLA(2)s) are water-soluble enzymes that bind to the surface of both artificial and biological lipid bilayers and hydrolyze the membrane phospholipids. The tissue expression pattern of the human group IID secretory phospholipase A(2) (hsPLA(2)-IID) suggests that the enzyme is involved in the regulation of the immune and inflammatory responses. With an aim to establish an expression system for the hsPLA(2)-IID in Escherichia coli, the DNA-coding sequence for hsPLA(2)-IID was subcloned into the vector pET3a, and expressed as inclusion bodies in E. coli (BL21). A protocol has been developed to refold the recombinant protein in the presence of guanidinium hydrochloride, using a size-exclusion chromatography matrix followed by dilution and dialysis to remove the excess denaturant. After purification by cation-exchange chromatography, far ultraviolet circular dichroism spectra of the recombinant hsPLA(2)-IID indicated protein secondary structure content similar to the homologous human group IIA secretory phospholipase A(2). The refolded recombinant hsPLA(2)-IID demonstrated Ca(2+)-dependent hydrolytic activity, as measuring the release free fatty acid from phospholipid liposomes. This protein expression and purification system may be useful for site-directed mutagenesis experiments of the hsPLA(2)-IID which will advance our understanding of the structure-function relationship and biological effects of the protein. (C) 2009 Elsevier Inc. All rights reserved.

STATIC IMAGES WITH DIFFERENT INDUCED INTENSITIES OF HUMAN BODY MOVEMENTS AFFECT SUBJECTIVE TIME

Modulation of subjective time was examined using static images eliciting perceptions of different intensities of body movement. Undergraduate students were exposed to photographs of dancer sculptures in different dance positions for 36 sec. and asked to estimate the exposure duration. Lower movement intensities were related to shorter estimated durations. Mean durations for images of unmoving dancers were underestimated and for dancers taking a ballet step were overestimated. Temporal estimations were also related to the order of presentation of the stimuli, which suggested that subjective time estimations were influenced by the experimental context. Subjective time is related not only to the visual perception of moving images, but also of elicited perceptions of movement in static images, suggesting an embodiment effect on subjective time estimation.

Impact of photodynamic therapy on inflammatory cells during human chronic periodontitis

The aim of this study was to evaluate the effects of the photodynamic therapy (PDT) on the inflammatory infiltrate and on the collagen network organization in human advanced chronic periodontitis Two different drug delivery systems (DDS) were tested (liposomes and nanoemulsions) to determine if the effects of PDT could differ according to the DDS used Sixteen patients presenting two teeth with chronic advanced periodontitis and Important tooth mobility with clinical indication of extraction were included in the group liposomes (group L n = 8) or in the group nanoemulsions (group N n = 8) in order to compare the effects of each DDS Seven days before extractions one tooth of each patient was treated with PDT using phthalocyanine derivatives as photosensitizers and the contralateral tooth was taken as control In group L the density of gingival collagen fibers (66 +/- 19%) was significantly Increased (p < 0 02) when compared to controls (35 +/- 21%) Concerning the antigen-presenting cells PDT had differential effects depending on the drug delivery system the number of macrophages was significantly decreased (p < 0 05) in group L while the number of Langerhans cells was significantly decreased in group N (p < 0 02) These findings demonstrate that PDT presents an impact on gingival Inflammatory phenomenon during chronic periodontitis and leads to a specific decrease of antigen-presenting cells populations according to the drug delivery system used (C) 2010 Elsevier B V All rights reserved

The Effect of Expectancy of a Threatening Event on Time Perception in Human Adults

Two experiments were conducted to investigate the effect of a threatening stimulus in human adults in a temporal bisection task. In Experiment I. for two anchor duration conditions (400/800 vs. 800/1600 ms), the participants completed trials in which the probe duration was followed by an aversive stimulus or a nonaversive stimulus. The results showed that the duration was judged longer when the participants expected an aversive rather than a nonaversive stimulus. In Experiment 2, the effect of the temporal localization of the aversive stimulus was also tested, with the aversive stimulus being presented at the beginning or at the end of the probe duration. The results revealed a temporal overestimation in each condition compared to the trials in which no aversive stimulus was presented. Furthermore, the temporal overestimation was greater when the expectation for the forthcoming threatening stimulus was longer. This temporal overestimation is explained in terms of a speeding-up of the neural timing system in response to the increase in the arousal level produced by the expectation of a threatening stimulus.

Preliminary study of human breast tissue using synchrotron radiation combining WAXS and SAXS techniques

Using synchrotron radiation, we combined simultaneously wide angle X-ray scattering (WAXS) and small angle X-ray scattering (SAXS) techniques to obtain the scattering profiles of normal and neoplastic breast tissu-es samples at the momentum transfer range 6.28 nm(-1) <= Q(=4 pi.sin(theta/2)lambda) <= 50.26 nm(-1) and 0.15 nm(-1) <= Q <= 1.90 nm(-1), respectively. The results obtained show considerable differences between the scattering profiles of these tissues. We verified that the combination of some parameters (ratio between glandular and adipose peak intensity and third-order axial peak intensity) extracted from scattering profiles can be used for identifying breast cancer. (c) 2009 Elsevier Ltd. All rights reserved.

Mapping of suramin binding sites on the group IIA human secreted phospholipase A(2)

Suramin is a polysulphonated napthylurea used as an antiprotozoal/anthelminitic drug, which also inhibits a broad range of enzymes. Suramin binding to recombinant human secreted group IIA phospholipase A(2) (hsPLA(2)GIIA) was investigated by molecular dynamics simulations (MD) and isothermal titration calorimetry (ITC). MD indicated two possible bound suramin conformations mediated by hydrophobic and electrostatic interactions with amino-acids in three regions of the protein. namely the active-site and residues located in the N- and C-termini, respectively. All three binding sites are located on the phospholipid membrane recognition surface, suggesting that suramin may inhibit the enzyme, and indeed a 90% reduction in hydrolytic activity was observed in the presence of 100 nM suramin. These results correlated with ITC data, which demonstrated 2.7 suramin binding sites on the hsPLA(2)GIIA, and indicates that suramin represents a novel class of phosphohpase A(2) inhibitor. (C) 2009 Elsevier Inc. All rights reserved.

Evaluation of the aquatic macroinvertebrate community as a tool for monitoring a reservoir in the Pitangui river basin, Parana, Brazil

Evaluation of the aquatic macroinvertebrate community as a tool for monitoring a reservoir in the Pitangui river basin, Parana, Brazil. Benthic and nektonic macroinvertebrates play an important role in the structure and function of aquatic ecosystems and their distribution is influenced by chemical features of the substrate, vegetation composition, and water depth. Knowledge on the fauna contributes to the assessment of water quality and development of biodiversity conservation activities. Different biotic factors affecting the invertebrate community were evaluated in the Alagados reservoir, the main water source of the city of Ponta Grossa, Parana. In five different sampling points, 18,473 specimens of aquatic or semiaquatic macroinvertebrates were collected, belonging to 46 taxa of the phylla Annelida (Hirudinea and Oligochaeta), Mollusca (Gastropoda), Platyhelminthes (Turbellaria), Nematoda and Arthropoda (Arachnida, Crustacea and Insecta). This community was composed mainly of predators (45.7% of the taxa sampled), collectors and/or filterers (23.9%), scrapers (15.2%), shredders (13.0%) and detritivores (2.2%). Diversity (H`) and evenness (J) indices were significantly low for the sites examined, and H` ranged between 0.3301 and 1.0396. Regarding tolerance of organisms to organic pollution, more sensitive taxa were very rare (Plecoptera) or unusual (Trichoptera and Ephemeroptera). Among the more resistant groups are Chironomidae and Hirudinea, both fairly common in the samples. This study corroborates the importance of bioindicators as a tool to assess water quality for human consumption and for the conservation of aquatic environments, integrating physical, chemical and biological factors in monitoring programs.