Eugenia uniflora: a promising natural alternative against multidrug-resistant bacteria.

This work aimed to evaluate the chemical composition, antioxidant and antimicrobial activities from crude extract and fractions from leaves of Eugenia uniflora Linn. The crude extract was obtained by turbo extraction and their fractions by partitioning. Chromatographic analysis were performed, and the antioxidant capacity was verified by two methods (DPPH• and ABTS•+). The Minimal Inhibitory/Bactericidal Concentration were conducted against twenty-two bacteria, selecting five strains susceptible to extract/fractions and resistant to the antibiotics tested. Ampicillin, azithromycin, ciprofloxacin, and gentamicin were associated with Ethyl Acetate Fraction (EAF) against multidrug-resistant strains in modulatory and checkerboard tests. The chromatographic data showed gallic acid, ellagic acid, and myricitrin in crude extract, with enrichment in the EAF. The electron transfer activity demonstrated in the antioxidant tests is related to the presence of flavonoids. The Gram-positive strains were more susceptible to EAF, and their action spectra were improved by association, comprising Gram-negative bacilli. Synergisms were observed to ciprofloxacin and gentamicin against Pseudomonas aeruginosa colistin-resistant. The results demonstrate that the extract and enriched fraction obtained from the leaves of E. uniflora act as a promising natural alternative against multidrug-resistant bacteria.

Eugenia uniflora: a promising natural alternative against multidrug-resistant bacteria / Eugenia uniflora: uma alternativa natural promissora contra bactérias multirresistentes

This work aimed to evaluate the chemical composition, antioxidant and antimicrobial activities from crude extract and fractions from leaves of Eugenia uniflora Linn. The crude extract was obtained by turbo extraction and their fractions by partitioning. Chromatographic analysis were performed, and the antioxidant capacity was verified by two methods (DPPH• and ABTS•+). The Minimal Inhibitory/Bactericidal Concentration were conducted against twenty-two bacteria, selecting five strains susceptible to extract/fractions and resistant to the antibiotics tested. Ampicillin, azithromycin, ciprofloxacin, and gentamicin were associated with Ethyl Acetate Fraction (EAF) against multidrug-resistant strains in modulatory and checkerboard tests. The chromatographic data showed gallic acid, ellagic acid, and myricitrin in crude extract, with enrichment in the EAF. The electron transfer activity demonstrated in the antioxidant tests is related to the presence of flavonoids. The Gram-positive strains were more susceptible to EAF, and their action spectra were improved by association, comprising Gram-negative bacilli. Synergisms were observed to ciprofloxacin and gentamicin against Pseudomonas aeruginosa colistin-resistant. The results demonstrate that the extract and enriched fraction obtained from the leaves of E. uniflora act as a promising natural alternative against multidrug-resistant bacteria.

Este trabalho teve como objetivo avaliar a composição química, as atividades antioxidantes e antimicrobianas do extrato bruto e frações de folhas de Eugenia uniflora Linn. O extrato bruto foi obtido por turbólise e suas frações por partição. Foram realizadas análises cromatográficas e a capacidade antioxidante foi verificada por dois métodos (DPPH• e ABTS•+). A Concentração Inibitória/Bactericida Mínima foi realizada contra vinte e duas bactérias, selecionando cinco cepas suscetíveis a extração/frações e resistentes aos antibióticos testados. Ampicilina, azitromicina, ciprofloxacina e gentamicina foram associados à Fração Acetato de Etila (FAE) contra cepas multirresistentes em testes modulatórios e de checkboard. Os dados cromatográficos mostraram ácido gálico, ácido elágico e miricitrina em extrato bruto, com enriquecimento na FAE. A atividade de transferência de elétrons demonstrada nos testes antioxidantes está relacionada com a presença de flavonoides. As cepas de Gram-positivas foram mais suscetíveis à FAE, e seus espectros de ação foram melhorados por associação, compreendendo bacilos Gram-negativos. Foram observados sinergismos de ciprofloxacina e gentamicina contra Pseudomonas aeruginosa resistente à colistina. Os resultados demonstram que o extrato e a fração enriquecida obtida das folhas de E. uniflora atuam como uma alternativa natural promissora contra bactérias multirresistentes.

Pathogenicity of Aphelenchoides Pseudogoodeyi on the Ornamental Plants Asplenium Nidus and Lilium Speciosum in Brazil.

Aphelenchoides pseudogoodeyi has recently been reported in association with seeds of forage grasses and rice in Brazil and senescent strawberry plants, in the United States. This nematode is likely a mycophagous species; however, so far, its pathogenicity potential to plants is unclear. This study aimed to verify the pathogenicity of A. pseudogoodeyi to two species of ornamental plants. The experiments were conducted by inoculating A. pseudogoodeyi onto Bird's-Nest Fern (Asplenium nidus) and Oriental Lily (Lilium speciosum) leaves, using two inoculation methods (with and without injury). After 40 days of inoculation (DAI) in Bird's-Nest Fern and 5, 10, 20 and 40 DAI in Oriental Lily, the pathogenicity and the host efficiency were evaluated by symptoms observation and by severity, final nematode population and reproductive factor (RF), respectively. Additionally, a histopathological study was performed by inoculating A. pseudogoodeyi onto Bird's-Nest Fern for observing anatomical alterations. A. pseudogoodeyi was able to cause local necrotic lesions on both Bird's-Nest Fern and Oriental Lily leaves. However, the presence of injury was essential to enable A. pseudogoodeyi to penetrate and cause those symptoms in both plant species. Also, the total population of A. pseudogoodeyi decreased drastically over time and RF was <1, which characterized these species as poor-host or resistant plants. A. pseudogoodeyi penetrated into the foliar tissue and induced a total destruction of the mesophyll and collapse of the cells, with the formation of large intercellular spaces. It is concluded that A. pseudogoodeyi is an opportunistic pathogen as injury was required to induce symptoms in Bird's-Nest Fern and Oriental Lily.

The effects of crocetin supplementation on the blastocyst outcome, transcriptomic and metabolic profile of in vitro produced bovine embryos.

The earliest stages of embryo development are deeply influenced by reactive oxygen species (ROS), byproducts of the mitochondrial oxygen metabolism that play a key role as messengers in normal cell signal transduction and cell cycling. Despite its positive roles, the imbalance caused by the excess of ROS and an inefficient antioxidant system leads to oxidative stress, with negative consequences to the cell such as DNA damage, metabolic changes, mitochondrial stress and cell death. In the present work, crocetin - a natural antioxidant - was added to the culture media of bovine embryos to evaluate the efficiency of its antioxidant capability during embryo culture. Oocytes were in vitro matured (IVM) and fertilized according to standard protocols. Embryos were cultured at 38.5 °C under humidified air with 5% CO2, 7% O2, and 90% N2 in Synthetic Oviduct Fluid (SOF) medium supplemented with amino acids and either 5% of FBS (SOFaa) (control group) or SOFaa supplemented with 1  µM crocetin (crocetin group). After 5 days from the beginning of in vitro culture (IVC) (day 5 - D5), embryos were transferred to individual drops of culture media. At day 7 (D7), embryos were assessed by means of blastocyst rates, morphophysiological analyzes (total cell number, ROS and mitochondrial activity levels), transcript quantitation of 47 genes and metabolomic evaluation of the culture media by Raman spectroscopy. In the crocetin group blastocyst rates were higher and embryos had increased total cell number and decreased intracellular levels of ROS. These embryos also had upregulation of genes related with response to stress and lipid metabolism (ATF4, BAX, FOXO3, GADD45A, GPX1, GPX4, HSF1, SOD2, ACACA, SREBF1 and SREBF2). Raman spectroscopy corroborated these results indicating more active lipid and amino acid production in this group. The absence of crocetin in the culture media resulted in higher ROS level, as well as up regulation of genes related to DNA damage, stress response and energy metabolism (MORF4L2, SOD1, TXN, PFKP, PGK1 and PPARGC1A). In conclusion, crocetin supplementation during culture protects embryos from oxidative stress and influences the adaptive response to stress conditions, leading to an increase in both blastocyst yield and quality, as well as changes in transcriptomic and metabolic profile of in vitro produced bovine embryos.

In Vitro and In Vivo Studies of the Trypanocidal Effect of Novel Quinolines.

Therapies for human African trypanosomiasis and Chagas disease, caused by Trypanosoma brucei and Trypanosoma cruzi, respectively, are limited, providing minimal therapeutic options for the millions of individuals living in very poor communities. Here the effects of 10 novel quinolines are evaluated in silico and by phenotypic studies using in vitro and in vivo models. Absorption, distribution, metabolism, excretion, and toxicity (ADMET) properties revealed that most molecules did not infringe on Lipinski's rules, which is a prediction of good oral absorption. These quinolines showed high probabilities of Caco2 permeability and human intestinal absorption and low probabilities of mutagenicity and of hERG1 inhibition. In vitro screens against bloodstream forms of T. cruzi demonstrated that all quinolines were more active than the reference drug (benznidazole [Bz]), except for DB2171 and DB2192, with five (DB2187, DB2131, DB2186, DB2191, and DB2217) displaying 50% effective concentrations (EC50s) of <3 µM (4-fold lower than that of Bz). Nine quinolines were more effective than Bz (2.7 µM) against amastigotes, showing EC50s ranging from 0.6 to 0.1 µM. All quinolines were also highly active in vitro against African trypanosomes, showing EC50s of ≤0.25 µM. The most potent and highly selective candidates for each parasite species were tested in in vivo models. Results for DB2186 were promising in mice with T. cruzi and T. brucei infections, reaching a 70% reduction of the parasitemia load for T. cruzi, and it cured 2 out of 4 mice infected with T. brucei DB2217 was also active in vivo and cured all 4 mice (100% cure rate) with T. brucei infection.

Melatonin reduces apoptotic cells, SOD2 and HSPB1 and improves the in vitro production and quality of bovine blastocysts.

Effects of adding different concentrations of melatonin (10-7 , 10-9 and 10-11  M) to maturation (Experiment 1; Control, IVM + 10-7 , IVM + 10-9 , IVM + 10-11 ) and culture media (Experiment 2; Control, IVC + 10-7 , IVC + 10-9 , IVC + 10-11 ) were evaluated on in vitro bovine embryonic development. The optimal concentration of melatonin (10-9  M) from Experiments 1-2 was tested in both maturation and/or culture media of Experiment 3 (Control, IVM + 10-9 , IVC + 10-9 , IVM/IVC + 10-9 ). In Experiment 1, maturated oocytes from Control and IVM + 10-9 treatments showed increased glutathione content, mitochondrial membrane potential and percentage of Grade I blastocysts (40.6% and 43%, respectively). In Experiment 2, an increase in the percentage of Grade I blastocysts was detected in IVC + 10-7 (43.5%; 56.7%) and IVC + 10-9 (47.4%; 57.4%). Moreover, a lower number and percentage of apoptotic cells in blastocysts were observed in the IVC + 10-9 group compared to Control (3.8 ± 0.6; 3.6% versus 6.1 ± 0.6; 5.3%). In Experiment 3, the IVC + 10-9 treatment increased percentage of Grade I blastocysts with a lower number of apoptotic cells compared to IVM/IVC + 10-9 group (52.6%; 3.0 ± 0.5 versus 46.0%; 5.4 ± 1.0). The IVC + 10-9 treatment also had a higher mRNA expression of antioxidant gene (SOD2) compared to the Control, as well as the heat shock protein (HSPB1) compared to the IVM + 10-9 . Reactive oxygen species production was greater in the IVM/IVC + 10-9 treatment group. In conclusion, the 10-9  M concentration of melatonin and the in vitro production phase in which it is used directly affected embryonic development and quality.

Supercritical Fluid Extraction of Pyrrolidine Alkaloid from Leaves of Piper amalago L.

Supercritical fluid extraction was used to extract the alkaloid N-[7-(3',4'-methylenedioxyphenyl)-2(Z),4(Z)-heptadienoyl]pyrrolidine from leaves of Piper amalago L. A three-level orthogonal array design matrix, OAD OA9(34), was used for optimization of the parameters of supercritical extraction of the alkaloid, employing supercritical carbon dioxide: extraction time (20, 40, and 60 min), temperature (40, 50, and 60°C), pressure (150, 200, and 250 bar), and the use of cosolvents (ethanol, methanol, and propyleneglycol). All parameters had significant effect on the alkaloid yield. The alkaloid yield after 60 min of extraction without cosolvents at 9 different conditions (32) in terms of temperature (40, 50, and 60°C) and pressure (150, 200, and 250 bar) was also evaluated. The optimal yield (≈3.8 mg g-1) was obtained with supercritical CO2 + methanol (5% v : v) at 40°C and 200 bar for 60 min of extraction.

Non-linear conduction due to depinning of charge order domains in Fe3O2BO3.

The oxyborate Fe3O2BO3 presents a charge density wave (CDW) transition close to room temperature. As we show here, this is associated with a well defined anomaly in the specific heat. Below this transition, when applying in a single crystal of Fe3O2BO3 a DC voltage above a temperature dependent threshold, a high current is liberated in this material. We study the conduction in single crystals of Fe3O2BO3 with voltage applied parallel and perpendicular to the crystallographic c axis direction. The observed currents are attributed to the depinning of charge ordered domains above a threshold voltage V T2 that gives rise to a collective conduction due to coherent domains. Compliance limited DC data shows that above a lower threshold voltage depinning is smooth and follows a power law scaling. Similar depinning with power law scaling is also revealed in the AC conductivity.

Phyllosphere Metaproteomes of Trees from the Brazilian Atlantic Forest Show High Levels of Functional Redundancy.

The phyllosphere of the Brazilian Atlantic Forest has been estimated to contain several million bacterial species that are associated with approximately 20000 plant species. Despite the high bacterial diversity in the phyllosphere, the function of these microorganisms and the mechanisms driving their community assembly are largely unknown. In this study, we characterized the bacterial communities in the phyllospheres of four tree species of the Atlantic Forest (Mollinedia schottiana, Ocotea dispersa, Ocotea teleiandra, and Tabebuia serratifolia) and their metaproteomes to examine the basic protein functional groups expressed in the phyllosphere. Bacterial community analyses using 16S rRNA gene sequencing confirmed prior observations that plant species harbor distinct bacterial communities and that plants of the same taxon have more similar communities than more distantly related taxa. Using LC-ESI-Q-TOF, we identified 216 nonredundant proteins, based on 3503 peptide mass spectra. Most protein families were shared among the phyllosphere communities, suggesting functional redundancy despite differences in the species compositions of the bacterial communities. Proteins involved in glycolysis and anaerobic carbohydrate metabolism, solute transport, protein metabolism, cell motility, stress and antioxidant responses, nitrogen metabolism, and iron homeostasis were among the most frequently detected. In contrast to prior studies on crop plants and Arabidopsis, a low abundance of OTUs related to Methylobacterium and no proteins associated with the metabolism of one-carbon molecules were detected in the phyllospheres of the tree species studied here. Our data suggest that even though the phyllosphere bacterial communities of different tree species are phylogenetically diverse, their metaproteomes are functionally convergent with respect to traits required for survival on leaf surfaces.

Morphokinetic-related response to stress in individually cultured bovine embryos.

The kinetics of in vitro-produced (IVP) bovine embryos is related to embryo viability, metabolism, and epigenetic patterns. Therefore, we believe that embryos with different speeds of development also respond differently to stress. In the present study, we performed global metabolic analysis (matrix-assisted laser desorption ionization time of flight mass spectrometry [MALDI-TOF]) of culture media, characterized apoptotic events (Terminal deoxynucleotidyl transferase dUTP nick end labeling [TUNEL] and caspase quantitation), and quantified transcript abundance of stress-related gene (real-time quantitative polymerase chain reaction [qRT-PCR]) in IVP bovine embryos with different developmental kinetics to investigate possible markers of stress response. For this purpose, embryos were considered "fast" if they presented four or more cells at 40 hours post insemination (hpi). Embryos presenting two cells at this time were classified as "slow". Evaluations were performed at 40 hpi, 112 hpi, and 186 hpi. Metabolome analysis revealed several metabolites differentially represented between groups at all time points related with energy, lipid and amino acids metabolism, and stress response. There was no difference in TUNEL positive cells between groups in any of the time points analyzed. Nevertheless, at 112 hpi, classified as a critical phase because of the genome activation, the amount of caspase 3 and 7 and total caspase were higher in slow when compared to fast group. Transcript abundance analysis of candidate genes (GRP78, HSP60, SOD1, and MORF4L2) was also different among groups. In conclusion, IVP bovine embryos of different development speeds respond differentially to the environmental stress leading to different metabolome patterns and apoptosis activation throughout the culture.

Rapid and noninvasive technique to assess the metabolomics profile of bovine embryos produced in vitro by Raman spectroscopy.

Morphological assessments are used to select embryos with the highest implantation potential, however it is still very limited. The development of new technologies, such as Raman spectroscopy have improved quantitative and qualitative analysis, and consequently led to a better characterization of embryos and improvements on the prediction of their potential. Therefore, we propose a method based on the conventional in vitro culture system of bovine embryos, and the subsequent analysis of the culture media drops by Raman spectroscopy. Our results obtained by PCA analysis clearly showed a separation of the spectral profiles from culture media drops with and without embryos.

Carotenoid consumption is related to lower lipid oxidation and DNA damage in middle-aged men.

The present cross-sectional study assessed the potential relationships of carotenoid intake with lipid and oxidative stress markers in middle-aged men. A total of 296 apparently healthy middle-aged men (mean age 50.5 (SD 5.0) years, BMI 25.8 (SD 3.5) kg/m(2)) were recruited to participate in the study. Dietary intake, anthropometry, blood pressure, lifestyle features, blood and urine biomarkers were assessed using validated procedures. The lipid markers included NEFA, Castelli index, and TAG:HDL ratio; oxidative stress markers included urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG), 8-iso-PGF2α and plasma oxidised-LDL (ox-LDL). We observed a significant inverse association (P < 0.05) between NEFA concentrations and consumption of lutein plus zeaxanthin, ß-carotene, α-carotene and total carotenoid, while Castelli index was negatively associated with daily intake of lycopene, ß-carotene and total carotenoids. Regarding oxidative stress biomarkers, urinary 8-OHdG and ox-LDL concentrations were also inversely associated (P < 0.05) with consumption of lycopene, lutein plus zeaxanthin, ß-carotene, α-carotene and total carotenoids, regardless of confounding variables. Moreover, there was a negative association of urinary 8-iso-PGF2α concentration with dietary lutein plus zeaxanthin (ß - 0.135, 95% CI - 0.268, - 0.001), ß-carotene (ß - 0.156, 95% CI - 0.277, - 0.034) and with the sum of all carotenoids (ß - 0.189, 95% CI - 0.333, - 0.046). In conclusion, total daily carotenoid intake based on five investigated carotenoid types (ß-cryptoxanthin, lycopene, lutein plus zeaxanthin, ß-carotene and α-carotene) was inversely associated with relevant lipid and oxidative stress markers in middle-aged men, with emphasis on ß-carotene that was negatively associated with five of the six lipid and oxidative stress markers evaluated in the present study.

Anxiolytic-like, stimulant and neuroprotective effects of Ilex paraguariensis extracts in mice.

Yerba-mate (Ilex paraguariensis St. Hil.) is the most used beverage in Latin America with approximately 426 thousand of tons consumed per year. Considering the broad use of this plant, we aimed to investigate the anxiety-like and stimulant activity of both the hydroethanolic (HE) and aqueous (AE) extracts from leaves of I. paraguariensis. Swiss mice were treated with I. paraguariensis HE or AE chronically or acutely, respectively, followed by evaluation in the elevated plus-maze (EPM; anxiety-like paradigm), open field (OF; locomotor activity) or the step-down avoidance task (memory assessment). Following behavioral protocols the brains were collected for evaluation of acetylcholinesterase (AChE) activity ex vivo. Chronic treatment with HE induced an anxiolytic-like effect and increased motor activity besides augmented AChE activity. Additionally, acute treatment with AE prevented the scopolamine-induced memory deficit in the step-down avoidance task. Overall, our results indicate the importance of the I. paraguariensis-induced CNS effects, since it is a widely used nutraceutical. We have reported anxiolytic, stimulant and neuroprotective effects for this plant species. These effects are potentially modulated by the cholinergic system as well as by caffeine.

Brilliant cresyl blue staining negatively affects mitochondrial functions in porcine oocytes.

The aim of the present study was to examine the effects of brilliant cresyl blue (BCB) staining on mitochondrial functions in porcine oocytes. Cumulus-oocyte complexes (COCs) collected from slaughterhouse-derived porcine ovaries were cultured with (13 µM) or without (0 µM, control) BCB for 60 min. Mitochondrial functions in oocytes were examined immediately after staining or after in vitro maturation. The BCB-stained oocytes produced reactive oxygen species (ROS) at higher levels than control oocytes immediately after staining (2.2-fold, P < 0.001) and after maturation (1.7-fold, P < 0.001). The adenosine triphosphate (ATP) content and mitochondrial membrane potential (MMP) in oocytes were similar for the two groups immediately after staining. However, ATP and relative MMP levels were significantly (P < 0.05) lower in BCB-treated oocytes than in the control (2.18 versus 2.83 pM and 0.82 versus 1.0, respectively). There was no difference in mitochondrial DNA copy number between the two groups after maturation. The ATP content in early developmental stage embryos (3 days after parthenogenetic activation) was lower in the BCB-stained group than that in the control group but the difference was not significant. In conclusion, BCB staining of oocytes at the immature stage compromises mitochondrial functions throughout oocyte maturation, but function is restored during early embryo development.

Proteome changes in Oncidium sphacelatum (Orchidaceae) at different trophic stages of symbiotic germination.

Mutualistic symbioses between plants and fungi are a widespread phenomenon in nature. Particularly in orchids, association with symbiotic fungi is required for seed germination and seedling development. During the initial stages of symbiotic germination, before the onset of photosynthesis, orchid protocorms are fully mycoheterotrophic. The molecular mechanisms involved in orchid symbiotic germination and development are largely unknown, but it is likely that changes in plant energy metabolism and defense-related responses play a central role in these processes. We have used 2D-LC-MS/MS coupled to isobaric tagging for relative and absolute quantification to identify proteins with differential accumulation in Oncidium sphacelatum at different stages of mycorrhizal protocorm development (achlorophyllous and green protocorms) after seed inoculation with a Ceratobasidium sp. isolate. We identified and quantified 88 proteins, including proteins putatively involved in energy metabolism, cell rescue and defense, molecular signaling, and secondary metabolism. Quantitative analysis showed that the expected changes in carbon metabolism in green protocorms were accompanied by enhanced accumulation of proteins involved in the modulation of reactive oxygen species homeostasis, defense-related responses, and phytoalexins and carotenoid biosynthesis. Our results suggest profound metabolic changes in orchid protocorms during the switch from the fully mycoheterotrophic to the photosynthetic stage. Part of these changes may be also related to the obligatory nature of the interaction with the endomycorrhizal fungus.

Meat and carcass traits of the red-winged tinamou (Rhynchotus rufescens)

Tinamou (Rhynchotus rufescens) is a native bird of the Brazilian fauna and has great potential to become a domesticated species. From this perspective, the objective of this study was to evaluate the carcass and main parts yield and to assess meat quality of that bird. Two trials, one studying stocking density and the other litter materials, lasting 60 days each, were conducted. The studied traits were evaluated in two birds/replicate/treatment. Performance was evaluated by the least square method. The results showed that bird age at slaughter significantly influenced carcass and breast yields (p<0.05). Mean carcass, breast, thighs+drumstick and wing yields were 86.50±2.78%, 32.84±1.50%, 27.71±1.44%, and 13.21±0.94%, respectively. Shear force, pH, water holding capacity, lightness, redness and yellowness means and standard deviations were 1.92±0.98 kgf.cm-2, 6.07±0.12; 70±0.53%, 48.30±0.96, 1.96±1.01, and 1.61±1.0, respectively. Moisture, ash, crude protein, fat, and cholesterol contents were 72.35±0.16%, 1.12±0.06%, 25.53±0.50%, 0.17±0.01%, and 18.87±2.76 mg/10g, respectively. Results showed carcass and parts yields and the physical-chemical traits of tinamou meat are similar to chicken meat; however, tinamou meat is less acidic and has higher protein level combined with less calories and lower cholesterol levels.(AU)

Chromosome number variation and evolution in Neotropical Leguminoseae (Mimosoideae) from northeastern Brazil.

Most members of the subfamily Mimosoideae have pantropical distributions, variable habits, and a basic chromosome number x = 13. We examined karyotypic evolution of 27 species of this subfamily occurring principally in northeastern Brazil by examining chromosomes stained with Giemsa. All of the species had semi-reticulated interphase nuclei and early condensing segments in the proximal region of both chromosome arms. The basic number x = 13 was the most frequent, with 2n = 2x = 26 in 19 of the species, followed by 2n = 4x = 52 and 2n = 6x = 78. However, the three species of the genus Calliandra had the basic number x = 8, with 2n = 2x = 16, while Mimosa cordistipula had 2n = 4x = 32. The karyotypes were relatively symmetrical, although bimodality was accentuated in some species, some with one or two acrocentric pairs. As a whole, our data support earlier hypotheses that the Mimosoideae subfamily has a basic number of x = 13 and underwent karyotypic evolution by polyploidy. However, x = 13 seems to be a secondary basic number that originated from an ancestral stock with x1 = 7, in which polyploidy followed by descending disploidy gave rise to the current lineages with x = 13. Another lineage, including current representatives of Calliandra with x = 8, may have arisen by ascending disploidy directly from an ancestral monoploid stock with x1 = 7.

Cryopreservation of immature equine oocytes, comparing a solid surface vitrification process with open pulled straws and the use of a synthetic ice blocker.

The objective was to evaluate the effect of three cryopreservation methods on the in vitro maturation (IVM) and membrane integrity (MIn) of immature equine oocytes. An open pulled straw (OPS) method, a novel solid surface vitrification (SSV) process, and the addition of a synthetic ice blocker were evaluated. Compared with the control group (N=269), the OPS (N=159) and the SSV (N=202) cryopreservation methods decreased both IVM (50.9 vs. 13.3 and 9.4%, respectively; P<0.001) and MIn (76.6 vs. 31.1 and 33.7%; P<0.001) of immature equine oocytes. However, inclusion of 0.1% ice blocker in the OPS vitrification process increased the rates of both IVM (30.5%; P<0.01) and MIn (45.8%; P<0.05) of the oocytes (N=59). Including 0.1% ice blocker in the SSV process improved the IVM rate (20.9%; P<0.05), whereas MIn remained compromised in this group (N=67). However, increasing the concentration of the ice blocker (to 1.0%) in the cryopreservation methods did not significantly improve rates of IVM. In conclusion, the addition of a synthetic ice blocker (0.1%) to both cryopreservation processes significantly increased rates of both IVM and MIn of immature equine oocytes cryopreserved by OPS.

Initial development of bovine placentation (Bos indicus) from the point of view of the allantois and amnion.

The aim of this study was to perform a morphological characterization of the initial bovine placental development, between 20 and 70 days post-insemination (p.i.), with emphasis on the differentiation of the allantois and amnion. After collection, the conceptuses were dissected, macroscopically measured and photographically documented. The extraembryonic membranes were cut into fragments measuring 5 cm(2), and then fixed in 4% paraformaldehyde for analysis using light microscopy, and in 2.5% glutaraldehyde for use in scanning and transmission electron microscopy. The extraembryonic and fetal membranes presented variable degrees of development throughout the periods analysed. The macroscopic appearance of vascularization of the allantois, its attempt to merge with the chorium and the effective appearance of the first cotyledons in development were the events observed from 30 to 40 days of pregnancy. The measurements of the amnion increased gradually as gestation developed. The allantoic epithelia presented cellular dimorphism from 20 to 25 days of pregnancy, but was shown to be immature from 60 to 70 days of pregnancy.

Lipoxygenase-derived eicosanoids are involved in the inhibitory effect of Crotalus durissus terrificus venom or crotoxin on rat macrophage phagocytosis.

Crotalus durissus terrificus snake venom and its major toxin, crotoxin or type II PLA2 subunit of this toxin, induce an inhibitory effect on spreading and phagocytosis in 2h incubated macrophages. The involvement of arachidonate-derived mediators on the inhibitory action of the venom or toxins on rat peritoneal macrophage phagocytosis was presently investigated. Peritoneal cells harvested from naive rats and incubated with the venom or toxins or harvested from the peritoneal cavity of rats pre-treated with the toxins were used. Zileuton, a 5-lipoxygenase inhibitor but not indomethacin, a cyclooxygenase inhibitor, given in vivo and in vitro abolished the inhibitory effect of venom or toxins on phagocytosis. Resident peritoneal macrophages incubated with the venom or toxins showed increased levels of prostaglandin E2 and lipoxin A4, with no change in leukotriene B4. These results suggest that lipoxygenase-derived eicosanoids are involved in the inhibitory effect of C.d. terrificus venom, crotoxin or PLA2 on macrophage phagocytosis.