Pollinator-mediated selection on Krameria oil flowers: A flower-pollinator fit adaptation to an atypical oil-collecting behaviour?

BACKGROUND AND AIMS: Spatial variation in plant-pollinator interactions is a key driver of floral trait diversification. A so far overlooked qualitative aspect of this variation is the behavioural component on flowers that relates to the pollinator fit. We tested the hypothesis that variation in pollinator behaviour influences the geographical pattern of phenotypic selection across the distribution range of the oil-producing Krameria grandiflora (Krameriaceae). This variation mainly involves the presence or absence of flag petal grasping, which is only performed by representatives of Centris (Centridini, Apidae), an oil-collecting bee group highly associated with Krameriaceae pollination. METHODS: We quantified variation in floral traits and fitness and estimated pollinator-mediated selection in five populations at a large geographical scale comprising the entire species range. In each population, we sampled individual pollen arrival and germination as a fitness measure, indicating pollination success and pollination performance, which was then relativized and regressed on standardized flower-pollinator fit (flag-stigma distance), advertisement (sepal length) and reward (oil volume) traits. This generated mean-scaled selection gradients used to calculate geographical selection dispersion. KEY RESULTS: Unexpectedly, stronger selection was detected on the flower-pollinator fit trait in populations highly associated to the absence of the flag petal grasping. Geographical variation in selection was mainly attributed to differential selection on the flag-stigma distance generating a selection mosaic. This may involve influences of a spatial variation in pollinator behaviour as well as composition and morphology. CONCLUSIONS: Our results show the adaptive significance of the specialized "flag" petals of Krameria in the absence of the grasping behaviour and highlight the contribution of geographical variation in pollinator behaviour on flowers in driving selection mosaics, with implications for floral evolution, adaptation to pollinator fit and phenotypic diversity in specialized systems.

Reproductive isolation between two sympatric bat-pollinated Bauhinia (Leguminosae).

Several barriers contribute to reproductive isolation between plant species, which can be classified as pre- or post-pollination. Understanding the strength of these barriers could clarify the factors that maintain reproductive isolation and thus species integrity. In this study, we quantified reproductive isolation between two bat-pollinated co-occurring Bauhinia species (B. acuruana and B. pentandra) with similar flower morphology. Over the course of 18 months, we assessed reproductive isolation between these two Bauhinia species by quantifying the individual strengths and absolute contributions of five pre- and post- pollination barriers. Our data showed that both species are completely isolated in their reproduction by a combination of several barriers. Although they co-occur in a few populations, we found a high degree of geographic isolation between them. And although their flowering periods overlap, there is a significant difference in flowering peaks. Both species have the same pollinating bats, but the interspecific transfer of pollen between the plant species may be reduced due to the different length of the flower stamens, resulting in different pollen deposition on the bats' bodies. We have documented complete incompatibility between taxa and conclude that pre- and post-pollination barriers are important factors in preventing gene flow, even in contact zones between these two species of Bauhinia. We highlight that our work is the first study to use methods to estimate the strength of reproductive isolation barriers between bat-pollinated species.

A negative association between nectar standing crop and pollen transfer suggests nectar functions as a manipulator of pollinating bats.

BACKGROUND AND AIMS: Nectar standing crop has a fundamental role in controlling pollinator movements between flowers and individuals within a population. In bat pollination systems, plants take advantage of the cognitive abilities of nectarivorous bats, which integrate complex perceptions of the quality and spatial distribution of resources. Here, we propose that associations between standing crop and pollen transfer help to reveal the role of nectar as a manipulator of pollinator behaviour. METHODS: We used Harpochilus neesianus Ness (Acanthaceae), a bat-pollinated shrub from the Brazilian Caatinga, as a model system to assess nectar removal effects and standing crop, respectively, over the night and to test associations between the amount of nectar available to pollinators, and pollen import and export. KEY RESULTS: Harpochilus neesianus showed continuous nectar secretion throughout the flower lifespan. Flowers subjected to successive nectar removals produced less nectar than flowers sampled just once, and showed, despite a higher sugar concentration, a lower absolute amount of sugar. Under these conditions, bats may realize that nectar production is decreasing after repeated visits to the same flower and could be manipulated to avoid such already pollinated flowers with little nectar, thus increasing the probability of visits to flowers with a high amount of nectar, and a still high pollen availability on anthers and low pollen deposition on stigmas. We found that during most of the period of anthesis, nectar standing crop volume was positively correlated with the number of pollen grains remaining in the anthers, and negatively with the number of pollen grains deposited on the stigma. CONCLUSIONS: Nectar secretion patterns can function as a manipulator of pollinating bats in H. neesianus. We propose that the assessment of variability in nectar secretion in response to removal, and the correlation between nectar standing crop and relative pollen transfer throughout anthesis should be considered in order to understand the role of nectar in the manipulation of pollinators.

The iconic cactus of the Caatinga dry forest, Cereus jamacaru (Cactaceae) has high sphingophily specialization and pollinator dependence.

Cereus jamacaru is a cactus distributed in Northeastern Brazil, with high symbolic value to this region. However, the interaction, behavior and the role of pollinators remains poorly understood. Here, we investigate the reproductive biology, addressing the ecological significance of floral attributes, including details about floral signaling. The study was carried at three areas of the Caatinga, in 2015, 2017 and 2021. We analyzed the floral morphometry, volume and concentration of the nectar, and characterized the colour and scent of flowers. Additionally, we described the pollinator behavior and performed controlled pollination experiments. The 'Mandacaru' is self-incompatible, has nocturnal anthesis and the nectar is accumulated as droplets in a long hypanthial tube. The flowers have a reflective pattern with a dark outer surface and a white inner surface. (E)-nerolidol is the major component (87.4%) of its floral perfume. We registered the sphingid moth Cocytius antaeus visiting the flowers. The floral attributes, attractants and rewards drives to a sphingophily, and the pollination treatments showed the dependence to fruit set by C. antaeus, the pollinator registered. In this case, if the apparent lack of pollinator diversity encompasses its entire range, the loss of the hawkmoth could severely impact the reproductive success of the cactus.

Porphyrin Macrocycles: General Properties and Theranostic Potential.

Despite specialists' efforts to find the best solutions for cancer diagnosis and therapy, this pathology remains the biggest health threat in the world. Global statistics concerning deaths associated with cancer are alarming; therefore, it is necessary to intensify interdisciplinary research in order to identify efficient strategies for cancer diagnosis and therapy, by using new molecules with optimal therapeutic potential and minimal adverse effects. This review focuses on studies of porphyrin macrocycles with regard to their structural and spectral profiles relevant to their applicability in efficient cancer diagnosis and therapy. Furthermore, we present a critical overview of the main commercial formulations, followed by short descriptions of some strategies approached in the development of third-generation photosensitizers.

Mixed pollination system and floral signals of Paepalanthus (Eriocaulaceae): insects and geitonogamy ensure high reproductive success.

BACKGROUND AND AIMS: Eriocaulaceae exhibit a great variety of floral traits associated with insect (e.g. nectariferous structures) and wind pollination (unisexual flowers, exposed sexual organs and small pollen grains), as well as the 'selfing syndrome' (small flowers, short distance between stigma and anthers, and temporal overlap of male and female phases). Paepalanthus bifidus, P. subtilis and P. tortilis are related species that differ in form, size and colour of floral structures. We aimed to investigate the pollination and reproductive biology of these three species. METHODS: We analysed the floral biology, floral visitors, pollinator behaviour, and the contribution of insects, wind and spontaneous geitonogamy to fruit set. We also evaluated the floral colour and scent of the species. Colour reflectance of capitula of each species was measured and plotted in models of insect vision. Floral scent samples were extracted and the compounds were compared to vegetative scent samples. KEY RESULTS: In all species, the staminate and pistillate flowers are arranged in alternating cycles with a temporal overlap between these phases. Ants were the most frequent floral visitors and were effective pollinators in P. bifidus and P. tortilis, while flies were occasional pollinators in P. tortilis. Floral visitors were not observed in P. subtilis. In all species, fruits were produced by spontaneous geitonogamy, with no evidence of wind pollination. According to the models of insect vision, the colours of the capitula of P. bifidus and P. subtilis are the most inconspicuous for ants and flies. We found no difference between the emission of volatiles of inflorescences and vegetative structures. CONCLUSIONS: This study suggests that ant pollination might be more widespread in Eriocaulaceae than currently assumed. Furthermore, for small monocarpic plants, mixed mating strategies are most favourable, by ensuring reproduction either by outcrossing when pollinators are abundant or by spontaneous geitonogamy when pollinations are scarce/absent.

Spectroscopic Analysis of Parathyroid and Thyroid Tissues by Ground-State diffuse Reflectance and Laser Induced Luminescence: a Preliminary Report.

Intraoperative discrimination of thyroid and parathyroid tissues is fundamental in thyroid surgery. Recent fluorescence studies have shown stronger NIR emission in parathyroid tissue than in thyroid tissue, presenting a potential avenue for the development of a tool for surgical assistance. However, the fluorophore responsible for this emission has not yet been identified. In this work, spectroscopic analysis was performed to ascertain the origin of the emission peaks in parathyroid tissue. Ground-state diffuse reflectance (GSDR) absorption spectroscopy and laser-induced luminescence (LIL) emission spectroscopy were performed in parathyroid, thyroid, and fatty tissue samples and the resulting spectra were compared with the peaks of known fluorophores to identify the origin of each peak. The spectra of the different tissue types were also compared in order to evaluate the wavelength which presents the highest parathyroid emission relative to the emission of the surrounding tissues, representing the ideal wavelength for parathyroid detection. An emission peak in these conditions was observed for both thyroid and parathyroid tissue at 711 nm, with a higher intensity in parathyroid sample, making it suitable for detection applications. These results show a potential avenue for the development of a system allowing parathyroid detection in a surgical setting.

Stress, anxiety and depression among gastronomes: association with workplace mobbing and work-family interaction.

INTRODUCTION: The work in commercial restaurant kitchens is characterized by many labor´s demands and strict commands that can cause damage to the mental health of gastronomes. The aim of this study was to verify the presence of stress, anxiety, and depression among gastronomy workers and associate them with the presence of mobbing at work. Still, it sought to evaluate the work-family interference and test its mediating role in the relationship between mobbing and psychopathology indicators. MATERIALS AND METHODS: This is an observational-analytical, cross-sectional study with a quantitative approach, in which 160 gastronomes participated (59.4% women), with an average age of 30.81 years. All answered a Sociodemographic and Labor Data Questionnaire, Negative Acts at Work Questionnaire, Depression Anxiety and Stress Scale, and Work-Family Interaction Scale, whose data were analyzed using descriptive and inferential statistics. RESULTS: A positive association was found between bullying and indicators of stress, anxiety, and depression, as well as negative interference between work-family and family-work with the same indicators. It was also observed the mediating effect of negative interference from work in family, in the relationship between mobbing and indicators of stress, anxiety, and depression. CONCLUSION: It was concluded, therefore, that the experience of mobbing affects the family and intensifies the psychological illness in gastronomes.

Flowers up! The effect of floral height along the shoot axis on the fitness of bat-pollinated species.

BACKGROUND AND AIMS: Bat-pollination is an important system in terms of occurrence and distribution, although it remains little studied. Thus, the role of particular flower traits in this interaction remains uncertain. Flower height along the shoot axis, associated with flower exposure, has often been deemed a key trait in this system, but its effect on fitness has not previously been assessed. We aimed to test its role and propose that taller flowers attain higher fitness due to a higher degree of accessibility and conspicuity to foraging bats. METHODS: We assessed the effect of floral height on bat visiting rates to individual flowers of Crescentia cujete (Bignoniaceae), a cauliflorous model bat-pollinated species with a marked gradient in flower height along the shoot axis. Additionally, we tested the effect of this variable on seed/ovule ratio measurements from seven other species from different families along a herb-tree gradient. Hypotheses were tested through mixed-effect linear models. KEY RESULTS: Bat visiting rates varied positively as a function of flower height in C. cujete, but significance was found only for the subset of flowers located on the trunk, closer to the ground. Similarly, seed/ovule ratios were positively correlated with flower height only for the three species with the shortest statures along the height gradient and shortest average floral heights. These results suggest that proximity to the ground, associated with herbaceous or bushy surrounding vegetation, may be an obstacle to the foraging of nectar-feeding bats, which in turn should explain the morphological convergence of inflorescence length and exposure strategies of short-statured bat-pollinated plants. CONCLUSIONS: Flower height has a species-specific effect on plant fitness. This study provides a novel numerical perspective to the role of an unexplored trait in bat-pollination, and has elucidated some aspects of the adaptive importance of flower height based on limitations imposed by ecologically complex pollinators.

The diversity and evolution of pollination systems in large plant clades: Apocynaceae as a case study.

Background and Aims: Large clades of angiosperms are often characterized by diverse interactions with pollinators, but how these pollination systems are structured phylogenetically and biogeographically is still uncertain for most families. Apocynaceae is a clade of >5300 species with a worldwide distribution. A database representing >10 % of species in the family was used to explore the diversity of pollinators and evolutionary shifts in pollination systems across major clades and regions. Methods: The database was compiled from published and unpublished reports. Plants were categorized into broad pollination systems and then subdivided to include bimodal systems. These were mapped against the five major divisions of the family, and against the smaller clades. Finally, pollination systems were mapped onto a phylogenetic reconstruction that included those species for which sequence data are available, and transition rates between pollination systems were calculated. Key Results: Most Apocynaceae are insect pollinated with few records of bird pollination. Almost three-quarters of species are pollinated by a single higher taxon (e.g. flies or moths); 7 % have bimodal pollination systems, whilst the remaining approx. 20 % are insect generalists. The less phenotypically specialized flowers of the Rauvolfioids are pollinated by a more restricted set of pollinators than are more complex flowers within the Apocynoids + Periplocoideae + Secamonoideae + Asclepiadoideae (APSA) clade. Certain combinations of bimodal pollination systems are more common than others. Some pollination systems are missing from particular regions, whilst others are over-represented. Conclusions: Within Apocynaceae, interactions with pollinators are highly structured both phylogenetically and biogeographically. Variation in transition rates between pollination systems suggest constraints on their evolution, whereas regional differences point to environmental effects such as filtering of certain pollinators from habitats. This is the most extensive analysis of its type so far attempted and gives important insights into the diversity and evolution of pollination systems in large clades.

Subtle Chemical Variations with Strong Ecological Significance: Stereoselective Responses of Male Orchid Bees to Stereoisomers of Carvone Epoxide.

Different enantiomers of chiral compounds within floral perfumes usually trigger distinct responses in insects; however, this has frequently been neglected in studies investigating semiochemicals in plant-pollinator interactions. Approximately 1000 neotropical plants produce floral perfumes as the only reward for pollinators, i.e. male euglossine bees. The chiral compound carvone epoxide is a key component of the scent bouquet of many perfume-rewarding plants that are pollinated by males of Eulaema. Here, we tested the biological activity of the four carvone epoxide stereoisomers to four Eulaema species occurring in the Atlantic Rainforest of NE-Brazil. We determined the stereochemistry of carvone epoxide in the floral scent of several Catasetum species, tested whether the antennae of bees respond differentially to these stereoisomers and investigated if there is a behavioural preference for any of the stereoisomers. We found that 1) Catasetum species emit only the (-)-trans-stereoisomer of carvone epoxide, 2) for E. atleticana and E. niveofasciata antennal responses to the (-)-trans-carvone epoxide were significantly stronger than those to (-)-cis-carvone epoxide, 3) the strength and pattern of antennal responses to all 4 stereoisomers (separately tested) did not differ among Eulaema species, and 4) there were significant differences in attractiveness of the four stereoisomers to the bees species with the (-)-trans-stereoisomer being particularly attractive. We assume (-)-trans-carvone epoxide to be the dominant isomer in perfume-rewarding plants pollinated by Eulaema. The universal occurrence of carvone epoxide in Catasetum species pollinated by Eulaema, suggests that this compound has evolved in perfume-rewarding as a specific attractant for Eulaema bees as pollinators.

Synthesis, Photochemical and In Vitro Cytotoxic Evaluation of New Iodinated Aminosquaraines as Potential Sensitizers for Photodynamic Therapy.

In this work, several benzothiazole-based aminosquaraine dyes, displaying strong absorption within the so-called phototherapeutic window (650⁻800 nm), were synthesized. The ability, of all the new dyes, to generate singlet oxygen was assessed by determining the correspondent phosphorescence emission and through the comparison with a standard. The quantum yields of singlet oxygen generation were determined and exhibited to be strongly dependent on the nature of the amino substituents introduced in the squaric ring. The photodynamic activity of the synthesized dyes was tested against four human tumor cell lines: breast (MCF-7), lung (NCI-H460), cervical (HeLa) and hepatocellular (HepG2) carcinomas; and a non-tumor porcine liver primary cell culture (PLP2). All the compounds synthesized were found to be able to inhibit tumor cells growth upon irradiation more than in the dark, in most of the cases, very significantly. Considering the photodynamic activity exhibited and the low toxicity displayed for the non-tumor cells, some of the synthetized dyes can be regarded as potential candidates as photosensitizers for PDT.

Endogenous annexin A1 (AnxA1) modulates early-phase gestation and offspring sex-ratio skewing.

Annexin A1 (AnxA1) is a glucocorticoid-regulated anti-inflammatory protein secreted by phagocytes and other specialised cells. In the endocrine system, AnxA1 controls secretion of steroid hormones and it is abundantly expressed in the testis, ovaries, placenta and seminal fluid, yet its potential modulation of fertility has not been described. Here, we observed that AnxA1 knockout (KO) mice delivered a higher number of pups, with a higher percentage of female offsprings. This profile was not dependent on the male features, as sperm from KO male mice did not present functional alterations, and had an equal proportion of Y and X chromosomes, comparable to wild type (WT) male mice. Furthermore, mismatched matings of male WT mice with female KO yielded a higher percentage of female pups per litter, a phenomenon which was not observed when male KO mice mated with female WT animals. Indeed, AnxA1 KO female mice displayed several differences in parameters related to gestation including (i) an arrested estrous cycle at proestrus phase; (ii) increased sites of implantation; (iii) reduced pre- and post-implantation losses; (iv) exacerbated features of the inflammatory reaction in the uterine fluid during implantation phase; and (v) enhanced plasma progesterone in the beginning of pregnancy. In summary, herein we highlight that AnxA1 pathway as a novel determinant of fundamental non-redundant regulatory functions during early pregnancy.

Studies on the Synthesis, Photophysical and Biological Evaluation of Some Unsymmetrical Meso-Tetrasubstituted Phenyl Porphyrins.

Abstract: We designed three unsymmetrical meso-tetrasubstituted phenyl porphyrins for further development as theranostic agents for cancer photodynamic therapy (PDT): 5-(4-hydroxy-3-methoxyphenyl)-10,15,20-tris-(4-acetoxy-3-methoxyphenyl)porphyrin (P2.2), Zn(II)-5-(4-hydroxy-3-methoxyphenyl)-10,15,20-tris-(4-acetoxy-3-methoxyphenyl)porphyrin (Zn(II)2.2) and Cu(II)-5-(4-hydroxy-3-methoxyphenyl)-10,15,20-tris-(4-acetoxy-3-methoxyphenyl)porphyrin (Cu(II)2.2). The porphyrinic compounds were synthesized and their structures were confirmed by elemental analysis, FT-IR, UV-Vis, EPR and NMR. The compounds had a good solubility in polar/nonpolar media. P2.2 and, to a lesser extent, Zn(II)2.2 were fluorescent, albeit with low fluoresence quantum yields. P2.2 and Zn(II)2.2 exhibited PDT-acceptable values of singlet oxygen generation. A "dark" cytotoxicity study was performed using cells that are relevant for the tumor niche (HT-29 colon carcinoma cells and L929 fibroblasts) and for blood (peripheral mononuclear cells). Cellular uptake of fluorescent compounds, cell viability/proliferation and death were evaluated. P2.2 was highlighted as a promising theranostic agent for PDT in solid tumors considering that P2.2 generated PDT-acceptable singlet oxygen yields, accumulated into tumor cells and less in blood cells, exhibited good fluorescence within cells for imagistic detection, and had no significant cytotoxicity in vitro against tumor and normal cells. Complexing of P2.2 with Zn(II) or Cu(II) altered several of its PDT-relevant properties. These are consistent arguments for further developing P2.2 in animal models of solid tumors for in vivo PDT.

Annexin A1 Is a Physiological Modulator of Neutrophil Maturation and Recirculation Acting on the CXCR4/CXCL12 Pathway.

Neutrophil production and traffic in the body compartments is finely controlled, and the strong evidences support the role of CXCL12/CXCR4 pathway on neutrophil trafficking to and from the bone marrow (BM). We recently showed that the glucocorticoid-regulated protein, Annexin A1 (AnxA1) modulates neutrophil homeostasis and here we address the effects of AnxA1 on steady-state neutrophil maturation and trafficking. For this purpose, AnxA1(-/-) and Balb/C wild-type mice (WT) were donors of BM granulocytes and mesenchymal stem cells and blood neutrophils. In vivo treatments with the pharmacological AnxA1 mimetic peptide (Ac2-26) or the formyl peptide receptor (FPR) antagonist (Boc-2) were used to elucidate the pathway of AnxA1 action, and with the cytosolic glucocorticoid antagonist receptor RU 38486. Accelerated maturation of BM granulocytes was detected in AnxA1(-/-) and Boc2-treated WT mice, and was reversed by treatment with Ac2-26 in AnxA1(-/-) mice. AnxA1 and FPR2 were constitutively expressed in bone marrow granulocytes, and their expressions were reduced by treatment with RU38486. Higher numbers of CXCR4(+) neutrophils were detected in the circulation of AnxA1(-/-) or Boc2-treated WT mice, and values were rescued in Ac2-26-treated AnxA1(-/-) mice. Although circulating neutrophils of AnxA1(-/-) animals were CXCR4(+) , they presented reduced CXCL12-induced chemotaxis. Moreover, levels of CXCL12 were reduced in the bone marrow perfusate and in the mesenchymal stem cell supernatant from AnxA1(-/-) mice, and in vivo and in vitro CXCL12 expression was re-established after Ac2-26 treatment. Collectively, these data highlight AnxA1 as a novel determinant of neutrophil maturation and the mechanisms behind blood neutrophil homing to BM via the CXCL12/CXCR4 pathway. J. Cell. Physiol. 231: 2418-2427, 2016. © 2016 Wiley Periodicals, Inc.

Chronic exposure of diesel exhaust particles induces alveolar enlargement in mice.

BACKGROUND: Diesel exhaust particles (DEPs) are deposited into the respiratory tract and are thought to be a risk factor for the development of diseases of the respiratory system. In healthy individuals, the timing and mechanisms of respiratory tract injuries caused by chronic exposure to air pollution remain to be clarified. METHODS: We evaluated the effects of chronic exposure to DEP at doses below those found in a typical bus corridor in Sao Paulo (150 µg/m3). Male BALB/c mice were divided into mice receiving a nasal instillation: saline (saline; n = 30) and 30 µg/10 µL of DEP (DEP; n = 30). Nasal instillations were performed five days a week, over a period of 90 days. Bronchoalveolar lavage (BAL) was performed, and the concentrations of interleukin (IL)-4, IL-10, IL-13 and interferon-gamma (INF-γ) were determined by ELISA-immunoassay. Assessment of respiratory mechanics was performed. The gene expression of Muc5ac in lung was evaluated by RT-PCR. The presence of IL-13, MAC2+ macrophages, CD3+, CD4+, CD8+ T cells and CD20+ B cells in tissues was analysed by immunohistochemistry. Bronchial thickness and the collagen/elastic fibers density were evaluated by morphometry. We measured the mean linear intercept (Lm), a measure of alveolar distension, and the mean airspace diameter (D0) and statistical distribution (D2). RESULTS: DEP decreased IFN-γ levels in BAL (p = 0.03), but did not significantly alter IL-4, IL-10 and IL-13 levels. MAC2+ macrophage, CD4+ T cell and CD20+ B cell numbers were not altered; however, numbers of CD3+ T cells (p ≤ 0.001) and CD8+ T cells (p ≤ 0.001) increased in the parenchyma. Although IL-13 (p = 0.008) expression decreased in the bronchiolar epithelium, Muc5ac gene expression was not altered in the lung of DEP-exposed animals. Although respiratory mechanics, elastic and collagen density were not modified, the mean linear intercept (Lm) was increased in the DEP-exposed animals (p ≤ 0.001), and the index D2 was statistically different (p = 0.038) from the control animals. CONCLUSION: Our data suggest that nasal instillation of low doses of DEP over a period of 90 days results in alveolar enlargement in the pulmonary parenchyma of healthy mice.

Aqueous extract of Rosmarinus officinalis L. inhibits neutrophil influx and cytokine secretion.

Rosmarinus officinalis L. phenolic compounds have attracted considerable attention because of their antioxidant and antimicrobial properties, including its ability to treat inflammatory disorders. In this work, we investigated the in vivo and in vitro effects of R. officinalis aqueous extract on neutrophil trafficking from the blood into an inflamed tissue, on cell-derived secretion of chemical mediators, and on oxidative stress. Anti-inflammatory activity was investigated using carrageenan-induced inflammation in the subcutaneous tissue of male Wistar rats orally treated with the R. officinalis extract (100, 200, or 400 mg/kg). The leukocyte influx (optical microscopy), secretion of chemical mediators (prostaglandin E2 (PGE2), TNF-α, interleukin 6 (IL-6), leukotriene B4 (LTB4), and cytokine-induced neutrophil chemoattractant 1 by enzyme-linked immunosorbent assay), and the anti-oxidative profile (super oxide dismutase (SOD), glutathione peroxidase, and thiobarbituric acid reactive substance (TBARS) spectrophotometry) were quantified in the inflamed exudate. N-Formyl-methionine-leucine-phenylalanine-induced chemotaxis, lipopolysaccharide-induced NO2 (-) production (Greiss reaction), and adhesion molecule expression (flow cytometry) were in vitro quantified using oyster glycogen recruited peritoneal neutrophils previous treated with the extract (1, 10, or 100 µg/mL). Animals orally treated with phosphate-buffered saline and neutrophils incubated with Hank's balanced salt solution were used as control. R. officinalis extract oral treatment caused a dose-dependent reduction in the neutrophil migration as well as decreased SOD, TBARS, LTB4, PGE2, IL-6, and TNF-α levels in the inflamed exudate. In vitro treatment with R. officinalis decreased neutrophil chemotaxis, NO2 (-) production, and shedding of L-selectin and ß2 integrin expressions. Results here presented show that R. officinalis aqueous extract displays important in vivo and in vitro anti-inflammatory actions by blocking pathways of neutrophil migration and secretion, suggesting its therapeutic application to acute inflammatory reactions.

Alterations in the profile of blood neutrophil membrane receptors caused by in vivo adrenocorticotrophic hormone actions.

Elevated levels of adrenocorticotrophic hormone (ACTH) mobilize granulocytes from bone marrow into the blood, although these neutrophils are refractory to a full migratory response into inflamed tissues. Here, we investigated the dependence of glucocorticoid receptor activation and glucocorticoid-regulated protein annexin A1 (ANXA1) on ACTH-induced neutrophilia and the phenotype of blood neutrophil after ACTH injection, focusing on adhesion molecule expressions and locomotion properties. ACTH injection (5 µg ip, 4 h) induced neutrophilia in wild-type (WT) mice and did not alter the elevated numbers of neutrophils in RU-38486 (RU)-pretreated or ANXA1(-/-) mice injected with ACTH. Neutrophils from WT ACTH-treated mice presented higher expression of Ly6G⁺ANXA1(high), CD18(high), CD62L(high), CD49(high), CXCR4(high), and formyl-peptide receptor 1 (FPR1(low)) than those observed in RU-pretreated or ANXA1(-/-) mice. The membrane phenotype of neutrophils collected from WT ACTH-treated mice was paralleled by elevated fractions of rolling and adherent leukocytes to the cremaster postcapillary venules together with impaired neutrophil migration into inflamed air pouches in vivo and in vitro reduced formyl-methionyl-leucyl-phenylalanine (fMLP) or stromal-derived factor-1 (SDF-1α)-induced chemotaxis. In an 18-h senescence protocol, neutrophils from WT ACTH-treated mice had a higher proportion of ANXAV(low)/CXCR4(low), and they were less phagocytosed by peritoneal macrophages. We conclude that alterations on HPA axis affect the pattern of membrane receptors in circulating neutrophils, which may lead to different neutrophil phenotypes in the blood. Moreover, ACTH actions render circulating neutrophils to a phenotype with early reactivity, such as in vivo leukocyte-endothelial interactions, but with impaired locomotion and clearance.

DNA aptamers are functional molecular recognition sensors in protic ionic liquids.

The function and structural changes of an AMP molecular aptamer beacon and its molecular recognition capacity for its target, adenosine monophosphate (AMP), was systematically explored in solution with a protic ionic liquid, ethylammonium nitrate (EAN). It could be proven that up to 2 M of EAN in TBS buffer, the AMP molecular aptamer beacon was still capable of recognizing AMP while also maintaining its specificity. The specificity was proven by using the guanosine monophosphate (GMP) as target; GMP is structurally similar to AMP but was not recognized by the aptamer. We also found that in highly concentrated EAN solutions the overall amount of double stranded DNA formed, as well as its respective thermal stability, diminished gradually, but surprisingly the hybridization rate (kh ) of single stranded DNA was significantly accelerated in the presence of EAN. The latter may have important implications in DNA technology for the design of biosensing and DNA-based nanodevices in nonconventional solvents, such as ionic liquids.

Wind pollination and propagule formation in Rhizophora mangle L. (Rhizophoraceae): resource or pollination limitation?

Rhizophora mangle is considered as a self-compatible mangrove, and is pollinated by wind and insects. However, there is no information about fruit production by autogamy and agamospermy and on the foraging behavior of its flower visitors. Hence, the present study analyzed the pollination and reproductive systems of R. mangle in a mangrove community in northern Pernambuco, Brazil. Floral morphology, sequence of anthesis, and behavior of flower visitors were described; the proportion of flowers that resulted in mature propagules was also recorded. Autogamy, agamospermy, and wind pollination tests were performed, and a new anemophily index is proposed. The flowers of R. mangle are hermaphrodite, protandric, and have high P/O rate. Flies were observed on flowers only during the male phase, probably feeding on mites that consume pollen. Rhizophora mangle is not agamospermic and its fruit production rate by spontaneous self-pollination is low (2.56%) compared to wind pollination (19.44%). The anemophily index was high 0.98, and thus it was considered as a good indicator. Only 13.79% of the flowers formed mature propagules. The early stages of fruit development are the most critical and susceptible to predation. Rhizophora mangle is, therefore, exclusively anemophilous in the study area and the propagule dispersal seems to be limited by herbivory.