Endothelium-Dependent Vasorelaxant Effect of Butanolic Fraction from Caryocar brasiliense Camb. Leaves in Rat Thoracic Aorta

Caryocar brasiliense Camb. "pequi" is a native plant from the Cerrado region of Brazil that contains bioactive components reported to be antioxidant agents. Previous work has demonstrated that dietary supplementation with pequi decreased the arterial pressure of volunteer athletes. We found that the crude hydroalcoholic extract (CHE) of C. brasiliense leaves relaxed, in a concentration-dependent manner, rat aortic rings precontracted with phenylephrine, and that the butanolic fraction (BF) produced an effect similar to that of the CHE. Aortic relaxation induced by BF was abolished by endothelium removal, by incubation of the nitric oxide synthase inhibitor L-NAME, or the soluble guanylatecyclase inhibitor ODQ. However, incubation with atropine and pyrilamine had no effect on the BF-induced vasorelaxation. Moreover, this effect was not inhibited by indomethacin and tetraethylammonium. The concentration-response curve to calcium in denuded-endothelium rings was not modified after incubation with BF, and the vasorelaxation by BF in endothelium-intact rings precontracted with KCl was abolished after incubation with L-NAME. In addition, administration of BF in anesthetized rats resulted in a reversible hypotension. The results reveal that C. brasiliense possesses both in vivo and in vitro activities and that the vascular effect of BF involves stimulation of the nitric oxide/cyclic GMP pathway.

Production of methyl and ethyl biodiesel fuel from pequi oil (Caryocar brasiliensis Camb.)

We studied the physical and chemical characteristics of methyl and ethyl esters (biodiesel) produced by transesterification of pequi oil (Caryocar brasiliensis Camb.) in the presence of potassium hydroxide. The oil extracted from pequi seed comprises 60% of the fruit content. Such characteristics as density, acidity, viscosity, and carbon residue of the biodiesel meet ANP (Brazilian National Petroleum Agency) standards. Our tests demonstrated the feasibility of utilizing pequi oil for biodiesel production.

Antigenotoxic Effects of Piquia (Caryocar villosum) in Multiple Rat Organs

This study investigated the in vivo genotoxicity of piquia pulp (Caryocar villosum) and its potential antigenotoxicity on doxorubicin (DXR)-induced DNA damage by comet assay and micronucleus test. In addition, the phytochemicals present in piquia pulp were determined. Piquia fruit pulp (75, 150 or 300 mg/kg b.w.) was administered by gavage to Wistar rats for 14 days, and the animals received an injection of saline or DXR (15 mg/kg b.w., i.p.) 24 h before they were euthanized. The phytochemical analysis revealed the presence of carotenoids; phenolic compounds, including flavonoids; tannins and alpha-tocopherol in piquia pulp. No statistically significant differences were observed in the evaluated parameters, demonstrating the absence of cytotoxic and genotoxic effects of piquia pulp at all tested doses. In liver, kidney, cardiac and bone marrow cells, piquia significantly reduced the DNA damage induced by DXR. Our results showed that the lowest piquia dose caused the largest decrease in DNA damage and the highest dose caused the smallest decrease, demonstrating an inverse dose-response of piquia pulp. Furthermore, we observed a difference in the potential antigenotoxic effects in several tissues. In conclusion, our results demonstrated that piquia pulp was not genotoxic and inhibited the genotoxicity induced by DXR, but some of the protective effects that were observed depended on the doses and experimental conditions. Therefore, further investigations are needed to clarify how piquia pulp positively affects human health.

Novel insights into the genomic basis of citrus canker based on the genome sequences of two strains of Xanthomonas fuscans subsp. aurantifolii

Abstract Background Citrus canker is a disease that has severe economic impact on the citrus industry worldwide. There are three types of canker, called A, B, and C. The three types have different phenotypes and affect different citrus species. The causative agent for type A is Xanthomonas citri subsp. citri, whose genome sequence was made available in 2002. Xanthomonas fuscans subsp. aurantifolii strain B causes canker B and Xanthomonas fuscans subsp. aurantifolii strain C causes canker C. Results We have sequenced the genomes of strains B and C to draft status. We have compared their genomic content to X. citri subsp. citri and to other Xanthomonas genomes, with special emphasis on type III secreted effector repertoires. In addition to pthA, already known to be present in all three citrus canker strains, two additional effector genes, xopE3 and xopAI, are also present in all three strains and are both located on the same putative genomic island. These two effector genes, along with one other effector-like gene in the same region, are thus good candidates for being pathogenicity factors on citrus. Numerous gene content differences also exist between the three cankers strains, which can be correlated with their different virulence and host range. Particular attention was placed on the analysis of genes involved in biofilm formation and quorum sensing, type IV secretion, flagellum synthesis and motility, lipopolysacharide synthesis, and on the gene xacPNP, which codes for a natriuretic protein. Conclusion We have uncovered numerous commonalities and differences in gene content between the genomes of the pathogenic agents causing citrus canker A, B, and C and other Xanthomonas genomes. Molecular genetics can now be employed to determine the role of these genes in plant-microbe interactions. The gained knowledge will be instrumental for improving citrus canker control.

Comparative genomic and transcriptome analyses of pathotypes of Xanthomonas citri subsp. citri provide insights into mechanisms of bacterial virulence and host range

Abstract Background Citrus bacterial canker is a disease that has severe economic impact on citrus industries worldwide and is caused by a few species and pathotypes of Xanthomonas. X. citri subsp. citri strain 306 (XccA306) is a type A (Asiatic) strain with a wide host range, whereas its variant X. citri subsp. citri strain Aw12879 (Xcaw12879, Wellington strain) is restricted to Mexican lime. Results To characterize the mechanism for the differences in host range of XccA and Xcaw, the genome of Xcaw12879 that was completed recently was compared with XccA306 genome. Effectors xopAF and avrGf1 are present in Xcaw12879, but were absent in XccA306. AvrGf1 was shown previously for Xcaw to cause hypersensitive response in Duncan grapefruit. Mutation analysis of xopAF indicates that the gene contributes to Xcaw growth in Mexican lime but does not contribute to the limited host range of Xcaw. RNA-Seq analysis was conducted to compare the expression profiles of Xcaw12879 and XccA306 in Nutrient Broth (NB) medium and XVM2 medium, which induces hrp gene expression. Two hundred ninety two and 281 genes showed differential expression in XVM2 compared to in NB for XccA306 and Xcaw12879, respectively. Twenty-five type 3 secretion system genes were up-regulated in XVM2 for both XccA and Xcaw. Among the 4,370 common genes of Xcaw12879 compared to XccA306, 603 genes in NB and 450 genes in XVM2 conditions were differentially regulated. Xcaw12879 showed higher protease activity than XccA306 whereas Xcaw12879 showed lower pectate lyase activity in comparison to XccA306. Conclusions Comparative genomic analysis of XccA306 and Xcaw12879 identified strain specific genes. Our study indicated that AvrGf1 contributes to the host range limitation of Xcaw12879 whereas XopAF contributes to virulence. Transcriptome analyses of XccA306 and Xcaw12879 presented insights into the expression of the two closely related strains of X. citri subsp. citri. Virulence genes including genes encoding T3SS components and effectors are induced in XVM2 medium. Numerous genes with differential expression in Xcaw12879 and XccA306 were identified. This study provided the foundation to further characterize the mechanisms for virulence and host range of pathotypes of X. citri subsp. citri.

Genome sequence of Xanthomonas fuscans subsp. fuscans strain 4834-R reveals that flagellar motility is not a general feature of xanthomonads

Abstract Background Xanthomonads are plant-associated bacteria responsible for diseases on economically important crops. Xanthomonas fuscans subsp. fuscans (Xff) is one of the causal agents of common bacterial blight of bean. In this study, the complete genome sequence of strain Xff 4834-R was determined and compared to other Xanthomonas genome sequences. Results Comparative genomics analyses revealed core characteristics shared between Xff 4834-R and other xanthomonads including chemotaxis elements, two-component systems, TonB-dependent transporters, secretion systems (from T1SS to T6SS) and multiple effectors. For instance a repertoire of 29 Type 3 Effectors (T3Es) with two Transcription Activator-Like Effectors was predicted. Mobile elements were associated with major modifications in the genome structure and gene content in comparison to other Xanthomonas genomes. Notably, a deletion of 33 kbp affects flagellum biosynthesis in Xff 4834-R. The presence of a complete flagellar cluster was assessed in a collection of more than 300 strains representing different species and pathovars of Xanthomonas. Five percent of the tested strains presented a deletion in the flagellar cluster and were non-motile. Moreover, half of the Xff strains isolated from the same epidemic than 4834-R was non-motile and this ratio was conserved in the strains colonizing the next bean seed generations. Conclusions This work describes the first genome of a Xanthomonas strain pathogenic on bean and reports the existence of non-motile xanthomonads belonging to different species and pathovars. Isolation of such Xff variants from a natural epidemic may suggest that flagellar motility is not a key function for in planta fitness.

Developmental basis of limb homology in Pleurodiran turtles, and the identity of the hooked element in the chelonian tarsus

Although Pleurodiran turtles represent an important component of extant turtle radiation, our knowledge of the development and homology of limb bones in turtles rests mostly upon observations made on derived members of the Cryptodiran clade. Herein, we describe limb development in three pleurodirans: Podocnemis unifilis Troschel, 1848, Podocnemis sextuberculata Cornalia, 1849 and Phrynops hilarii (Dumeril and Bibron, 1835), in an effort to contribute to filling this anatomical gap. For earlier stages of limb development, we described the Y-shaped condensation that gave rise to the zeugopodial cartilages, and differentiation of the primary axis/digital arch that reveals the invariant pattern common to tetrapods. There are up to four central cartilaginous foci in the carpus, and the proximal tarsale is formed by the fusion of the fibulare, intermedium, and centrale 4. Digital development is similar for the five digits. Changes in toe V occur predominantly in the distal tarsale 5. Ontogenetic reduction of phalanges is observed in toe V of Podocnemis. Based on these results, we suggest that the hooked element present in the chelonian tarsus, and traditionally recognized as a modified fifth metatarsale, is actually the fifth distal tarsale. Additionally, our data on limb development of pleurodiran turtles supply more taxonomically comprehensive information to interpret limb configuration within the chelonian clade. (C) 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 155, 845-866.

Growth, organic acids profile and sugar metabolism of Bifidobacterium lactis in co-culture with Streptococcus thermophilus: the inulin effect

The organic acids profile, sugar metabolism and biomass growth of Streptococcus thermophilus (St) and Bifidobacterium lactis (BI) have been studied in pure cultures or binary co-culture (St-BI) in skim milk either containing 40 mg/g of inulin or not. With inulin, the time required by St. BI and St-BI to complete fermentation (i.e., when the pH reached 4.5) was about 14, 8 and 49% shorter than without inulin, respectively. This prebiotic also enhanced the levels of lactic and acetic acids and volatile compounds, showing a positive synbiotic effect between pre- and probiotics. In particular, the St-BI co-culture showed final concentrations of both microorganisms about 15 and 38% higher than in their respective pure cultures, thus highlighting a clear synergistic effect between these microorganisms due to mutual interactions. In addition, the well-known bifidogenic effect of inulin was confirmed. (c) 2012 Elsevier Ltd. All rights reserved.

A synopsis and notes for Baccharis subgen. Tarchonanthoides (Asteraceae: Astereae)

Nomenclatural and taxonomic notes are provided as the result of a taxonomic revision of Baccharis subgen. Tarchonanthoides, currently underway. A list of accepted species within the subgenus, their synonyms, and a revised sectional classification are presented. Protologues and types of previously published names have been reviewed. Currently 21 species and 10 synonyms are recognized within the subgenus. Morphologically, the species of the subgenus are classified into four sections: Canescentes (9 spp.), Coridifoliae (8 spp.), Curitybensis (2 spp.), and Tarchonanthoides (2 spp.). A new combination and status is proposed to B. coridifolia subsp. bicolor, B. uleana is assigned to subgen. Tarchonanthoides, and lectotypes are designated for B. artemisioides, B. curitybensis, B. erigeroides var. dusenii, B. gibertii, B. patens, B. psammophila and B. squarrosa, most of them providing 'second-step' lectotypes.

LIZARDS AND SNAKES (LEPIDOSAURIA, SQUAMATA) FROM THE LATE QUATERNARY OF THE STATE OF CEARA IN NORTHEASTERN BRAZIL

We present the first formal report on the squamate assemblage from Parque Nacional de Ubajara. This park contains the most important cave complex in the state of Ceara in northeastern Brazil, called Provincia Espeleologica de Ubajara. The material comes from the Urso Fossil cave at Pendurado Hill. All previously reported fossil remains found in this cave are tentatively attributed to the Quaternary (late Pleistocene-early Holocene). Probably only Arctotherium brasiliense represents a relictual fossil bear from the late Pleistocene megafauna. The taxa recognized in this paper belong to Tropidurus sp., Ameiva sp., cf. Epicrates, and cf. Crotalus durissus, adding to the knowledge of the Brazilian Quaternary squamate fauna as a whole, and contribute to a major taxonomic refinement of the squamate assemblages from the early Holocene of northeastern Brazil.

Fatty acid profile, trans-octadecenoic, alpha-linolenic and conjugated linoleic acid contents differing in certified organic and conventional probiotic fermented milks

Development of dairy organic probiotic fermented products is of great interest as they associate ecological practices and benefits of probiotic bacteria. As organic management practices of cow milk production allow modification of the fatty acid composition of milk (as compared to conventional milk), we studied the influence of the type of milk on some characteristics of fermented milks, such as acidification kinetics. bacterial counts and fatty acid content. Conventional and organic probiotic fermented milks were produced using Bifidobacterium animalis subsp. lactis HN019 in co-culture with Streptococcus thermophilus TA040 and Lactobacillus delbrueckii subsp. bulgaricus LB340. The use of organic milk led to a higher acidification rate and cultivability of Lactobacillus bulgaricus. Fatty acids profile of organic fermented milks showed higher amounts of trans-octadecenoic acid (C18:1, 1.6 times) and polyunsaturated fatty acids, including cis-9 trans-11. C18:2 conjugated linoleic (CLA-1.4 times), and alpha-linolenic acids (ALA-1.6 times), as compared to conventional fermented milks. These higher levels were the result of both initial percentage in the milk and increase during acidification, with no further modification during storage. Finally, use of bifidobacteria slightly increased CLA relative content in the conventional fermented milks, after 7 days of storage at 4 degrees C, whereas no difference was seen in organic fermented milks. (C) 2012 Elsevier Ltd. All rights reserved.

Fibers from fruit by-products enhance probiotic viability and fatty acid profile and increase CLA content in yoghurts

This study evaluated the effect of the supplementation of total dietary fiber from apple, banana or passion fruit processing by-products on the post-acidification, total titratable acidity, bacteria counts and fatty acid profiles in skim milk yoghurts co-fermented by four different probiotics strains: Lactobacillus acidophilus L10 and Bifidobacterium animalis subsp. lactis BL04, HN019 and B94. Apple and banana fibers increased the probiotic viability during shelf-life. All the fibers were able to increase the short chain and polyunsaturated fatty acid contents of yoghurts compared to their respective controls. A synergistic effect between the type of fiber and the probiotic strain on the conjugated linoleic acid content was observed, and the amount of alpha-linolenic acid was increased by banana fiber. The results of this study demonstrate, for the first time, that fruit fibers can improve the fatty acid profile of probiotic yoghurts and point out the suitability of using fibers from fruit processing the by-products to develop new high value-added fermented dairy products. (C) 2012 Elsevier By. All rights reserved.

Organic milk improves Bifidobacterium lactis counts and bioactive fatty acids contents in fermented milk

In functional dairy products, polyunsaturated fatty acids such as, conjugated linoleic acid (CLA) and alpha-linolenic acid (ALA) have been highlighted for their benefits related to prevention of some chronic diseases. In order to study the effect of type of milk (conventional vs. organic, characterized by a specific fatty acid composition), Bifidobacterium animalis subsp. lactis (BB12, B94, BL04 and HN019) counts, acidification activity and chemical composition (pH, lactose, lactic acid contents and fatty acids profile) were investigated before fermentation and after 24 h of products stored at 4 degrees C. Organic and conventional milk influenced acidification performance and bacteria counts, which was strain-dependent. Higher counts of BB12 were observed in organic milk, whereas superior counts of BL04 were found in conventional milk. Organic fermented milk showed lower levels in saturated fatty acids (FA) and higher in monounsaturated FA contents. Similarly, among bioactive FA, organic fermented milks have higher amounts of trans vaccenic acid (TVA-C18:1t), conjugated linoleic acid (CLA) and slightly higher contents of alpha-linoleic acid (ALA). (C) 2012 Elsevier Ltd. All rights reserved.

Evaluation of different selective media for enumeration of probiotic micro-organisms in combination with yogurt starter cultures in fermented milk

The aim of this study was to assess selective plating methodologies for the enumeration and identification of Streptococcus thermophilus, Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus acidophilus, Lactobacillus rhamnosus and Bifidobacterium animalis ssp. lactis in fermented milks. Seven agar media (MRS with added sorbitol, clindamycin or vancomycin, acidified MRS, RCA with added aniline blue and dicloxacilin, M17 and ST) were evaluated. The results showed that RCA dicloxacilin agar was suitable for the selective enumeration of B. animalis ssp. lactis in fermented milk. Either MRS (acidified) or M17 agar could be used for enumeration of L. delbrueckii ssp. bulgaricus and S. thermophilus, respectively. MRS media containing antibiotics were effective for the enumeration of the probiotic organisms (L. rhamnosus and L. acidophilus) inoculated in fermented milks.

Dried Tomato-Flavored Probiotic Cream Cheese with Lactobacillus paracasei

A dried tomato-flavored probiotic cream cheese (P) containing Lactobacillus paracasei Lpc-37 was developed for the purpose of this study. The same product, but without probiotic addition (C) was used as control. Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris were used as lactic starter cultures. Chemical composition analyses and sensory tests were performed on days 1 and 7, respectively. Titratable acidity, pH value and L. paracasei population were determined every 7 d during the refrigerated storage (21 d) of the cream cheeses. The experiment and analyses were performed in triplicate, using standard methods. Probiotic population remained greater than 10(7) CFU/g throughout the storage period, thereby characterizing the product as potentially probiotic. Cream cheeses C and P did not differ on the sensory tests, both obtaining good overall acceptance by the consumers, of which 82.6% stated that they certainly or probably would buy the product.