Science.gov

Sample records for hevea brasiliensis clone

  1. Cloning and characterisation of JAZ gene family in Hevea brasiliensis.

    PubMed

    Hong, H; Xiao, H; Yuan, H; Zhai, J; Huang, X

    2015-05-01

    Mechanical wounding or treatment with exogenous jasmonates (JA) induces differentiation of the laticifer in Hevea brasiliensis. JA is a key signal for latex biosynthesis and wounding response in the rubber tree. Identification of JAZ (jasmonate ZIM-domain) family of proteins that repress JA responses has facilitated rapid progress in understanding how this lipid-derived hormone controls gene expression and related physiological processes in plants. In this work, the full-length cDNAs of six JAZ genes were cloned from H. brasiliensis (termed HbJAZ). These HbJAZ have different lengths and sequence diversity, but all of them contain Jas and ZIM domains, and two of them contain an ERF-associated amphiphilic repression (EAR) motif in the N-terminal. Real-time RT-PCR analyses revealed that HbJAZ have different expression patterns and tissue specificity. Four HbJAZ were up-regulated, one was down-regulated, while two were less effected by rubber tapping treatment, suggesting that they might play distinct roles in the wounding response. A yeast two-hybrid assay revealed that HbJAZ proteins interact with each other to form homologous or heterogeneous dimer complexes, indicating that the HbJAZ proteins may expand their function through diverse JAZ-JAZ interactions. This work lays a foundation for identification of the JA signalling pathway and molecular mechanisms of latex biosynthesis in rubber trees. PMID:25399518

  2. Molecular cloning and characterization of a Mlo gene in rubber tree (Hevea brasiliensis).

    PubMed

    Qin, Bi; Zheng, Fucong; Zhang, Yu

    2015-03-01

    Mlo gene encodes a plant-specific seven-transmembrane domain protein involved in a variety of cellular processes. In this study, a novel Mlo gene from rubber tree (Hevea brasiliensis), designated HbMlo1, was cloned by RT-PCR in rubber tree. The ORF of HbMlo1 was 1551bp in length, encoding a putative protein of 516 amino acids. HbMlo1 was a typical Mlo protein with seven-transmembrane domain. Sequence comparison between HbMlo1 and other Mlo proteins demonstrated that HbMlo1 shared the highest similarity with the Cucumis melo CmMlo1 and Arabidopsis thaliana AtMlo1 with 75.1% and 71.3% sequence identity, respectively. Phylogenetic analysis revealed that HbMlo1, CmMlo1, AtMlo1, AtMlo13, and AtMlo15 formed into the phylogenetic clade II with 100% bootstrap support value. HbMlo1 transcript exhibited tissue specificity, and it was preferentially expressed in leaf. Furthermore, the amount of HbMlo1 transcript was significantly induced by various phytohormones (including ethephon, methyl jasmonate, salicylic acid, abscisic acid, indole-3-acetic acid, and gibberellic acid), H2O2, and wounding treatments. Under drought stress, HbMlo1 exhibited a complex pattern of regulation. However, HbMlo1 expression did not significantly change during powdery mildew infection. These results suggested that HbMlo1 might play a role in phytohormone signaling and abiotic stress response processes in rubber tree. PMID:25506769

  3. Molecular cloning, expression profiles and characterization of a glutathione reductase in Hevea brasiliensis.

    PubMed

    Deng, Zhi; Zhao, Manman; Liu, Hui; Wang, Yuekun; Li, Dejun

    2015-11-01

    Glutathione reductase (GR; EC 1.8.1.7) is an important oxidoreductase that can protect organisms against various oxidative stresses. In this study, a new GR gene, named as HbGR2, was isolated from Hevea brasiliensis. The HbGR2 cDNA contained a 1674-bp open reading frame encoding 557 amino acids and the deduced HbGR2 protein showed high identities to the chloroplastic GRs from other plant species. HbGR2 was localized in the chloroplasts of tobacco mesophyll protoplasts. The cis-acting regulatory elements related to stress or hormone responses were predicted in the promoter region of HbGR2. The results from real-time RT-PCR analyses showed that HbGR2 was expressed throughout different tissues and developmental stages of leaves. Besides being related to tapping panel dryness (TPD), HbGR2 was regulated by several treatments including ethephon (ET), methyl jasmonate (MeJA), drought, low temperature, high salt, wounding and hydrogen peroxide (H2O2). The Escherichia coli (E. coli) cells overexpressing HbGR2 markedly increased their tolerance and survival at high concentrations of H2O2, suggesting that HbGR2 might play an important role in oxidative stress response in Hevea brasiliensis. PMID:26232647

  4. Comparative Transcriptome Analysis of Latex Reveals Molecular Mechanisms Underlying Increased Rubber Yield in Hevea brasiliensis Self-Rooting Juvenile Clones

    PubMed Central

    Li, Hui-Liang; Guo, Dong; Zhu, Jia-Hong; Wang, Ying; Chen, Xiong-Ting; Peng, Shi-Qing

    2016-01-01

    Rubber tree (Hevea brasiliensis) self-rooting juvenile clones (JCs) are promising planting materials for rubber production. In a comparative trial between self-rooting JCs and donor clones (DCs), self-rooting JCs exhibited better performance in rubber yield. To study the molecular mechanism associated with higher rubber yield in self-rooting JCs, we sequenced and comparatively analyzed the latex of rubber tree self-rooting JCs and DCs at the transcriptome level. Total raw reads of 34,632,012 and 35,913,020 bp were obtained from the library of self-rooting JCs and DCs, respectively, by using Illumina HiSeq 2000 sequencing technology. De novo assemblies yielded 54689 unigenes from the library of self-rooting JCs and DCs. Among 54689 genes, 1716 genes were identified as differentially expressed between self-rooting JCs and DCs via comparative transcript profiling. Functional analysis showed that the genes related to the mass of categories were differentially enriched between the two clones. Several genes involved in carbohydrate metabolism, hormone metabolism and reactive oxygen species scavenging were up-regulated in self-rooting JCs, suggesting that the self-rooting JCs provide sufficient molecular basis for the increased rubber yielding, especially in the aspects of improved latex metabolisms and latex flow. Some genes encoding epigenetic modification enzymes were also differentially expressed between self-rooting JCs and DCs. Epigenetic modifications may lead to gene differential expression between self-rooting JCs and DCs. These data will provide new cues to understand the molecular mechanism underlying the improved rubber yield of H. brasiliensis self-rooting clones. PMID:27555864

  5. Comparative Transcriptome Analysis of Latex Reveals Molecular Mechanisms Underlying Increased Rubber Yield in Hevea brasiliensis Self-Rooting Juvenile Clones.

    PubMed

    Li, Hui-Liang; Guo, Dong; Zhu, Jia-Hong; Wang, Ying; Chen, Xiong-Ting; Peng, Shi-Qing

    2016-01-01

    Rubber tree (Hevea brasiliensis) self-rooting juvenile clones (JCs) are promising planting materials for rubber production. In a comparative trial between self-rooting JCs and donor clones (DCs), self-rooting JCs exhibited better performance in rubber yield. To study the molecular mechanism associated with higher rubber yield in self-rooting JCs, we sequenced and comparatively analyzed the latex of rubber tree self-rooting JCs and DCs at the transcriptome level. Total raw reads of 34,632,012 and 35,913,020 bp were obtained from the library of self-rooting JCs and DCs, respectively, by using Illumina HiSeq 2000 sequencing technology. De novo assemblies yielded 54689 unigenes from the library of self-rooting JCs and DCs. Among 54689 genes, 1716 genes were identified as differentially expressed between self-rooting JCs and DCs via comparative transcript profiling. Functional analysis showed that the genes related to the mass of categories were differentially enriched between the two clones. Several genes involved in carbohydrate metabolism, hormone metabolism and reactive oxygen species scavenging were up-regulated in self-rooting JCs, suggesting that the self-rooting JCs provide sufficient molecular basis for the increased rubber yielding, especially in the aspects of improved latex metabolisms and latex flow. Some genes encoding epigenetic modification enzymes were also differentially expressed between self-rooting JCs and DCs. Epigenetic modifications may lead to gene differential expression between self-rooting JCs and DCs. These data will provide new cues to understand the molecular mechanism underlying the improved rubber yield of H. brasiliensis self-rooting clones. PMID:27555864

  6. Cloning and characterization of HbMT2a, a metallothionein gene from Hevea brasiliensis Muell. Arg differently responds to abiotic stress and heavy metals

    SciTech Connect

    Li, Yan; Chen, Yue Yi; Yang, Shu Guang; Tian, Wei Min

    2015-05-22

    Metallothioneins (MTs) are of low molecular mass, cysteine-rich proteins. They play an important role in the detoxification of heavy metals and homeostasis of intracellular metal ions, and protecting against intracellular oxidative damages. In this study a full-length cDNA of type 2 plant metallothioneins, HbMT2a, was isolated from 25 mM Polyethyleneglycol (PEG) stressed leaves of Hevea brasiliensis by RACE. The HbMT2a was 372 bp in length and had a 237 bp open reading frame (ORF) encoding for a protein of 78 amino acid residues with molecular mass of 7.772 kDa. The expression of HbMT2a in the detached leaves of rubber tree clone RY7-33-97 was up-regulated by Me-JA, ABA, PEG, H{sub 2}O{sub 2}, Cu{sup 2+} and Zn{sup 2+}, but down-regulated by water. The role of HbMT2a protein in protecting against metal toxicity was demonstrated in vitro. PET-28a-HbMT2-beared Escherichia coli. Differential expression of HbMT2a upon treatment with 10 °C was observed in the detached leaves of rubber tree clone 93-114 which is cold-resistant and Reken501 which is cold-sensitive. The expression patterns of HbMT2a in the two rubber tree clones may be ascribed to a change in the level of endogenous H{sub 2}O{sub 2}. - Highlights: • Cloning an HbMT2a gene from rubber tree. • Analyzing expression patterns of HbMT2a upon abiotic stress and heavy metal stress. • Finding different expression patterns of HbMT2a among two Hevea germplasm. • The expressed protein of HbMT2a enhances copper and zinc tolerance in Escherichia coli.

  7. Molecular cloning and characterization of a novel bi-functional α-amylase/subtilisin inhibitor from Hevea brasiliensis.

    PubMed

    Bunyatang, Orawan; Chirapongsatonkul, Nion; Bangrak, Phuwadol; Henry, Robert; Churngchow, Nunta

    2016-04-01

    A novel cDNA encoding a bi-functional α-amylase/subtilisin inhibitor (HbASI) was isolated from rubber (Hevea brasiliensis) leaves cultivar RRIM600. The HbASI had strong homology with the soybean trypsin inhibitor (Kunitz) family of protease inhibitors. Its putative amino acid sequence was similar to that of the α-amylase/subtilisin inhibitor from Ricinus communis (72% identity). Genomic sequencing indicated that the HbASI gene contained no introns. The messenger RNA of HbASI was detected in leaf, hypocotyl and root. The recombinant HbASI expressed extracellularly in Pichia pastoris exhibited inhibitory activity against α-amylase from Aspergillus oryzae, trypsin and subtilisin A. The HbASI gene was induced in the rubber leaves infected with a rubber tree pathogen, Phytophthora palmivora. It was also enhanced by salicylic acid (SA) treatment and mechanical wounding. In addition, the biological activity of the HbASI protein involving in the plant defence responses was also investigated. The HbASI at a concentration of 0.16 mg mL(-1) could inhibit the mycelium growth of P. palmivora. These data suggested that the HbASI protein might play a crucial role in defence against pathogen of rubber trees. PMID:26854410

  8. Cloning, heterologous expression and characterization of ascorbate peroxidase (APX) gene in laticifer cells of rubber tree (Hevea brasiliensis Muell. Arg.).

    PubMed

    Chao, Jinquan; Zhang, Shixin; Chen, Yueyi; Tian, Wei-Min

    2015-12-01

    Ascorbate peroxidases (APXs) are a kind of crucial enzymes for removing reactive oxygen species (ROS) in plant cell. In the present study, a full-length cDNA encoding an APX, designated HbAPX, was isolated from Hevea brasiliensis by the rapid amplification of cDNA ends (RACE) method. HbAPX was 1174-bp in length and contained a 912-bp open reading frame (ORF) encoding a putative protein of 304 amino acids. The predicted molecular mass of HbAPX was 27.6 kDa (kDa) with an isoelectric point (pI) of 6.73. The phylogenetic analysis showed that HbAPX belonged to the cytosolic subgroup and was more relative to PtAPX and MdAPX2. By using PlantCare online analysis, such cis-acting elements as W-box and MRE were detected in the promoter region of HbAPX. Overproduction of recombinant HbAPX protein either in Escherichia coli or yeast enhanced their tolerance to such abiotic stresses as Cu(2+), Zn(2+), Na(2+) and hydrogen peroxide (H2O2). Ethrel application significantly down-regulated the expression of HbAPX and inhibited the activity of HbAPX in vivo. The ethrel-caused down-regulation of HbAPX may disturb the redox homeostasis in laticifer cells of rubber tree. PMID:26519821

  9. Cloning and characterization of HbMT2a, a metallothionein gene from Hevea brasiliensis Muell. Arg differently responds to abiotic stress and heavy metals.

    PubMed

    Li, Yan; Chen, Yue Yi; Yang, Shu Guang; Tian, Wei Min

    2015-05-22

    Metallothioneins (MTs) are of low molecular mass, cysteine-rich proteins. They play an important role in the detoxification of heavy metals and homeostasis of intracellular metal ions, and protecting against intracellular oxidative damages. In this study a full-length cDNA of type 2 plant metallothioneins, HbMT2a, was isolated from 25 mM Polyethyleneglycol (PEG) stressed leaves of Hevea brasiliensis by RACE. The HbMT2a was 372bp in length and had a 237bp open reading frame (ORF) encoding for a protein of 78 amino acid residues with molecular mass of 7.772 kDa. The expression of HbMT2a in the detached leaves of rubber tree clone RY7-33-97 was up-regulated by Me-JA, ABA, PEG, H2O2, Cu(2+) and Zn(2+), but down-regulated by water. The role of HbMT2a protein in protecting against metal toxicity was demonstrated in vitro. PET-28a-HbMT2-beared Escherichia coli. Differential expression of HbMT2a upon treatment with 10 °C was observed in the detached leaves of rubber tree clone 93-114 which is cold-resistant and Reken501 which is cold-sensitive. The expression patterns of HbMT2a in the two rubber tree clones may be ascribed to a change in the level of endogenous H2O2. PMID:25858315

  10. Comparative study on the technological properties of latex and natural rubber from Hancornia speciosa Gomes and Hevea brasiliensis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This work reports a systematic comparative study of the properties of natural lattices and rubbers extracted from Hancornia speciosa Gomes and Hevea brasiliensis [(Willd. ex Adr. de Juss.) Muell.-Arg.] (clone RRIM 600) trees from 11 collections in Brazil throughout 2004. Natural rubber latex particl...

  11. Genetic and epigenetic uniformity of polyembryony derived multiple seedlings of Hevea brasiliensis.

    PubMed

    Karumamkandathil, Rekha; Uthup, Thomas K; Sankaran, Sobha; Unnikrishnan, Divya; Saha, Thakurdas; Nair, Sushamakumari S

    2015-05-01

    Hevea brasiliensis Muell. Arg (Para rubber tree) is a tropical tree species of Amazonian origin widely cultivated in several parts of the world for natural rubber, a highly priced commodity inevitable for the world rubber industry. Large, tree to tree variation in growth and latex yield among individual plants of high yielding Hevea clones is a common phenomenon observed in mature rubber plantations. The genetic heterogeneity of the seedlings which are used as rootstocks for propagation through budgrafting is considered as a major factor responsible for this variation. In order to minimize this variation, attempts were made to develop highly uniform rootstock material via an in vitro technique by inducing zygotic polyembryony in Hevea. Immature open pollinated fruits of a high yielding clone RRII 105 were cultured by half ovulo embryo culture technique. Multiple embryos were induced from the 8-10-week-old zygote with a novel combination of gibberellic acid (GA3), kinetin, and zeatin. Plantlets were successfully generated from the multiple embryos and raised in the field post hardening. Screening using genetic and epigenetic molecular markers revealed that the multiple seedlings developed are highly uniform and are of single zygotic origin. Development of plants having genetic and epigenetic uniformity suggests that this technique is ideal for raising uniform rootstock material in Hevea which may significantly reduce intraclonal variations. Moreover, these plants could serve as ideal material for physiological and molecular investigations towards the understanding of stock-scion interaction process in rubber. PMID:25359186

  12. Influence of soil, plant and meteorological factors on water relations and yield in Hevea brasiliensis

    NASA Astrophysics Data System (ADS)

    Rao, G. Gururaja; Rao, P. Sanjeeva; Rajagopal, R.; Devakumar, A. S.; Vijayakumar, K. R.; Sethuraj, M. R.

    1990-09-01

    Influence of factors governing the soil-plantatmosphere system on components of water relations and yield was studied in two clones of rubber tree, Hevea brasiliensis, viz. RRII 105 and RRII 118. Clonal variations were evident in yield and yield components and associated physiological parameters in response to soil moisture status and meteorological factors. Observations made during different seasons indicatedvariations in yield are attributed to differences in plugging index and initial flow rates, to the major yield components and also variations in components of water relations as influenced by meteorological factors. Among the two clones, RRII 105 was found to be fairly drought tolerant compared to RRII 118. RRII 105 was found to respond well to dry weather through higher stomatal resistances, higher leaf water potentials, lowered transpirational water loss and lower relative transpiration ratios, while RRII 118 was susceptible to stress situations.

  13. Involvement of Ethylene in the Latex Metabolism and Tapping Panel Dryness of Hevea brasiliensis

    PubMed Central

    Putranto, Riza-Arief; Herlinawati, Eva; Rio, Maryannick; Leclercq, Julie; Piyatrakul, Piyanuch; Gohet, Eric; Sanier, Christine; Oktavia, Fetrina; Pirrello, Julien; Kuswanhadi; Montoro, Pascal

    2015-01-01

    Ethephon, an ethylene releaser, is used to stimulate latex production in Hevea brasiliensis. Ethylene induces many functions in latex cells including the production of reactive oxygen species (ROS). The accumulation of ROS is responsible for the coagulation of rubber particles in latex cells, resulting in the partial or complete stoppage of latex flow. This study set out to assess biochemical and histological changes as well as changes in gene expression in latex and phloem tissues from trees grown under various harvesting systems. The Tapping Panel Dryness (TPD) susceptibility of Hevea clones was found to be related to some biochemical parameters, such as low sucrose and high inorganic phosphorus contents. A high tapping frequency and ethephon stimulation induced early TPD occurrence in a high latex metabolism clone and late occurrence in a low latex metabolism clone. TPD-affected trees had smaller number of laticifer vessels compared to healthy trees, suggesting a modification of cambial activity. The differential transcript abundance was observed for twenty-seven candidate genes related to TPD occurrence in latex and phloem tissues for ROS-scavenging, ethylene biosynthesis and signalling genes. The predicted function for some Ethylene Response Factor genes suggested that these candidate genes should play an important role in regulating susceptibility to TPD. PMID:26247941

  14. Involvement of Ethylene in the Latex Metabolism and Tapping Panel Dryness of Hevea brasiliensis.

    PubMed

    Putranto, Riza-Arief; Herlinawati, Eva; Rio, Maryannick; Leclercq, Julie; Piyatrakul, Piyanuch; Gohet, Eric; Sanier, Christine; Oktavia, Fetrina; Pirrello, Julien; Kuswanhadi; Montoro, Pascal

    2015-01-01

    Ethephon, an ethylene releaser, is used to stimulate latex production in Hevea brasiliensis. Ethylene induces many functions in latex cells including the production of reactive oxygen species (ROS). The accumulation of ROS is responsible for the coagulation of rubber particles in latex cells, resulting in the partial or complete stoppage of latex flow. This study set out to assess biochemical and histological changes as well as changes in gene expression in latex and phloem tissues from trees grown under various harvesting systems. The Tapping Panel Dryness (TPD) susceptibility of Hevea clones was found to be related to some biochemical parameters, such as low sucrose and high inorganic phosphorus contents. A high tapping frequency and ethephon stimulation induced early TPD occurrence in a high latex metabolism clone and late occurrence in a low latex metabolism clone. TPD-affected trees had smaller number of laticifer vessels compared to healthy trees, suggesting a modification of cambial activity. The differential transcript abundance was observed for twenty-seven candidate genes related to TPD occurrence in latex and phloem tissues for ROS-scavenging, ethylene biosynthesis and signalling genes. The predicted function for some Ethylene Response Factor genes suggested that these candidate genes should play an important role in regulating susceptibility to TPD. PMID:26247941

  15. Assembly and Analysis of Differential Transcriptome Responses of Hevea brasiliensis on Interaction with Microcyclus ulei

    PubMed Central

    Restrepo Restrepo, Silvia; Aristizábal Gutiérrez, Fabio Ancizar; Montoya Castaño, Dolly

    2015-01-01

    Natural rubber (Hevea brasiliensis) is a tropical tree used commercially for the production of latex, from which 40,000 products are generated. The fungus Microcyclus ulei infects this tree, causing South American leaf blight (SALB) disease. This disease causes developmental delays and significant crop losses, thereby decreasing the production of latex. Currently several groups are working on obtaining clones of rubber tree with durable resistance to SALB through the use of extensive molecular biology techniques. In this study, we used a secondary clone that was resistant to M. ulei isolate GCL012. This clone, FX 3864 was obtained by crossing between clones PB 86 and B 38 (H. brasiliensis x H. brasiliensis). RNA-Seq high-throughput sequencing technology was used to analyze the differential expression of the FX 3864 clone transcriptome at 0 and 48 h post infection (hpi) with the M. ulei isolate GCL012. A total of 158,134,220 reads were assembled using the de novo assembly strategy to generate 90,775 contigs with an N50 of 1672. Using a reference-based assembly, 76,278 contigs were generated with an N50 of 1324. We identified 86 differentially expressed genes associated with the defense response of FX 3864 to GCL012. Seven putative genes members of the AP2/ERF ethylene (ET)-dependent superfamily were found to be down-regulated. An increase in salicylic acid (SA) was associated with the up-regulation of three genes involved in cell wall synthesis and remodeling, as well as in the down-regulation of the putative gene CPR5. The defense response of FX 3864 against the GCL012 isolate was associated with the antagonistic SA, ET and jasmonic acid (JA) pathways. These responses are characteristic of plant resistance to biotrophic pathogens. PMID:26287380

  16. Assembly and Analysis of Differential Transcriptome Responses of Hevea brasiliensis on Interaction with Microcyclus ulei.

    PubMed

    Hurtado Páez, Uriel Alonso; García Romero, Ibonne Aydee; Restrepo Restrepo, Silvia; Aristizábal Gutiérrez, Fabio Ancizar; Montoya Castaño, Dolly

    2015-01-01

    Natural rubber (Hevea brasiliensis) is a tropical tree used commercially for the production of latex, from which 40,000 products are generated. The fungus Microcyclus ulei infects this tree, causing South American leaf blight (SALB) disease. This disease causes developmental delays and significant crop losses, thereby decreasing the production of latex. Currently several groups are working on obtaining clones of rubber tree with durable resistance to SALB through the use of extensive molecular biology techniques. In this study, we used a secondary clone that was resistant to M. ulei isolate GCL012. This clone, FX 3864 was obtained by crossing between clones PB 86 and B 38 (H. brasiliensis x H. brasiliensis). RNA-Seq high-throughput sequencing technology was used to analyze the differential expression of the FX 3864 clone transcriptome at 0 and 48 h post infection (hpi) with the M. ulei isolate GCL012. A total of 158,134,220 reads were assembled using the de novo assembly strategy to generate 90,775 contigs with an N50 of 1672. Using a reference-based assembly, 76,278 contigs were generated with an N50 of 1324. We identified 86 differentially expressed genes associated with the defense response of FX 3864 to GCL012. Seven putative genes members of the AP2/ERF ethylene (ET)-dependent superfamily were found to be down-regulated. An increase in salicylic acid (SA) was associated with the up-regulation of three genes involved in cell wall synthesis and remodeling, as well as in the down-regulation of the putative gene CPR5. The defense response of FX 3864 against the GCL012 isolate was associated with the antagonistic SA, ET and jasmonic acid (JA) pathways. These responses are characteristic of plant resistance to biotrophic pathogens. PMID:26287380

  17. Microsatellite marker development for the rubber tree (Hevea brasiliensis): characterization and cross-amplification in wild Hevea species

    PubMed Central

    2012-01-01

    Background The rubber tree (Hevea brasiliensis) is native to the Amazon region and it is the major source of natural rubber in the world. Rubber tree breeding is time-consuming and expensive. However, molecular markers such as microsatellites can reduce the time required for these programs. This study reports new genomic microsatellite markers developed and characterized in H. brasiliensis and the evaluation of their transferability to other Hevea species. Findings We constructed di- and trinucleotide-enriched libraries. From these two libraries, 153 primer pairs were designed and initially evaluated using 9 genotypes of H. brasiliensis. A total of 119 primer pairs had a good amplification product, 90 of which were polymorphic. We chose 46 of the polymorphic markers and characterized them in 36 genotypes of H. brasiliensis. The expected and observed heterozygosities ranged from 0.1387 to 0.8629 and 0.0909 to 0.9167, respectively. The polymorphism information content (PIC) values ranged from 0.097 to 0.8339, and the mean number of alleles was 6.4 (2–17). These 46 microsatellites were also tested in 6 other Hevea species. The percentage of transferability ranged from 82% to 87%. Locus duplication was found in H. brasiliensis and also in 5 of other species in which transferability was tested. Conclusions This study reports new microsatellite markers for H. brasiliensis that can be used for genetic linkage mapping, quantitative trait loci identification and marker- assisted selection. The high percentage of transferability may be useful in the evaluations of genetic variability and to monitor introgression of genetic variability from different Hevea species into breeding programs. PMID:22731927

  18. Molecular evolution and functional characterisation of haplotypes of an important rubber biosynthesis gene in Hevea brasiliensis.

    PubMed

    Uthup, T K; Rajamani, A; Ravindran, M; Saha, T

    2016-07-01

    Hydroxy-methylglutaryl coenzyme-A synthase (HMGS) is a rate-limiting enzyme in the cytoplasmic isoprenoid biosynthesis pathway leading to natural rubber production in Hevea brasiliensis (rubber). Analysis of the structural variants of this gene is imperative to understand their functional significance in rubber biosynthesis so that they can be properly utilised for ongoing crop improvement programmes in Hevea. We report here allele richness and diversity of the HMGS gene in selected popular rubber clones. Haplotypes consisting of single nucleotide polymorphisms (SNPs) from the coding and non-coding regions with a high degree of heterozygosity were identified. Segregation and linkage disequilibrium analysis confirmed that recombination is the major contributor to the generation of allelic diversity, rather than point mutations. The evolutionarily conserved nature of some SNPs was identified by comparative DNA sequence analysis of HMGS orthologues from diverse taxa, demonstrating the molecular evolution of rubber biosynthesis genes in general. In silico three-dimensional structural studies highlighting the structural positioning of non-synonymous SNPs from different HMGS haplotypes revealed that the ligand-binding site on the enzyme remains impervious to the reported sequence variations. In contrast, gene expression results indicated the possibility of association between specific haplotypes and HMGS expression in Hevea clones, which may have a downstream impact up to the level of rubber production. Moreover, haplotype diversity of the HMGS gene and its putative association with gene expression can be the basis for further genetic association studies in rubber. Furthermore, the data also show the role of SNPs in the evolution of candidate genes coding for functional traits in plants. PMID:26787454

  19. Biological control of Rigidoporus lignosus in Hevea brasiliensis in Nigeria.

    PubMed

    Ogbebor, Nicholas Obehi; Adekunle, Adefunke Temitayo; Eghafona, Odeh Nosakhare; Ogboghodo, Abraham Ikponmwosa

    2015-01-01

    The study was conducted to evaluate in vitro and in vivo control of fungal antagonists on Rigidoporus lignosus (klotzsch) Imaz in Hevea brasiliensis (Willd. ex A. Juss.) Muell. Arg. in the rain forest zone of Nigeria. In vitro assessments of biological agents were carried out in dixenic cultures in Petri plates. In vivo tests were carried out in the nursery using Hypocrea virens and Hypocrea jecorina. Significant differences were observed with the dual inoculations of fungal antagonists and R. lignosus inoculated the same day, and the antagonists inoculated 24 h before inoculation of R. lignosus. Hypocrea jecorina was most effective in the control of R. lignosus with percentage inhibition of 86.83 %. Hypocrea virens, H. jecorina, Trichoderma spirale, Trichoderma sp., Trichoderma sp. Pers, and Hypocrea lixii were effective on R. lignosus. Fungal antagonists inoculated 24 h before inoculation of R. lignosus exhibited higher inhibitory efficacies than antagonists and R. lignosus inoculated the same day. In the in vivo evaluation of fungi antagonists on R. lignosus, seedlings in H. jecorina treatment at 60 d and H. virens treatment at 150 d after inoculations respectively had the highest plant heaths (highest length of stem and length of tap rot, lowest plant death, and least foliar symptom) compared to the control. Mortality rate was higher at 60 d after inoculation and decline from the third months onward. Significant differences were observed between the control and R. lignosus treatment in all the parameters evaluated. PMID:25601145

  20. Highlights on Hevea brasiliensis (pro)hevein proteins.

    PubMed

    Berthelot, Karine; Peruch, Frédéric; Lecomte, Sophie

    2016-08-01

    Hevein, from Hevea brasiliensis (rubber tree), was identified in 1960. It is the most abundant soluble protein (22%) found in latex. Hevein is formed from a larger protein called prohevein. The 187 amino-acid prohevein is cleaved into two fragments: the N-terminal 43 amino-acid hevein, a lectin bearing a chitin-binding motif with antifungal properties, and a C-terminal domain (C-ter), which possesses amyloid properties. Hevein-like proteins are also widely represented in the plant kingdom and belong to a larger family related to stress and pathogenic responses. During the last 55 years, these proteins have attracted the interest of numerous specialists from the fields of plant physiology, genetics, molecular and structural biology, and physico-chemistry to allergology. This review highlights various aspects of hevein, prohevein, and C-ter from the point of view of these various fields, and examines their potential roles in latex as well as their beneficial and negative biological effects (e.g. wound sealing and resistance to pathogens which is mediated by agglutination, antimicrobial activity, and/or allergenicity). It covers results and observations from 1960 up to the most recent research. PMID:27317253

  1. Hydrophobic allergens from the bottom fraction membrane of Hevea brasiliensis.

    PubMed

    Mengumpun, Kesajee; Tayapiwatana, Chatchai; Hamilton, Robert G; Sangsupawanich, Pasuree; Wititsuwannakul, Rapepun

    2008-01-01

    Several proteins of rubber latex have been recognized as allergens causing immediate hypersensitivity in humans. In this study, a bottom fraction membrane (BFM) protein preparation from Hevea brasiliensis trees grown in southern Thailand was used to detect specific IgE in four groups of serum samples. The first group included 170 samples of latex glove factory workers (LGWs); group 2 consisted of the sera of 35 health care workers (HCWs) who were repeatedly exposed to powdered latex gloves; groups 3 and 4 were 31 positive and 22 negative sera, respectively, obtained from Johns Hopkins University School of Medicine, Baltimore, USA, tested for IgE to latex allergen. It was found that 56/170 (33%), 5/35 (14%), 11/31 (35.5%) and 1/22 (4.5%) samples of the LGWs, HCWs, CAP+ and CAP- groups had significant IgE to the BFM proteins, respectively. However, of all subjects only one subject of group 1 had experienced allergic morbidity consisting of eczema, conjunctivitis and asthma. The IgE of this subject bound to a 55 kDa component in the rubber latex BFM preparation. Thus, this protein may be regarded as a novel, although minor, latex allergen. Further investigation is needed to characterize the component and to pinpoint its allergenic role. PMID:19054931

  2. Variation in GUS activity in vegetatively propagated Hevea brasiliensis transgenic plants.

    PubMed

    Lardet, Ludovic; Leclercq, Julie; Bénistan, Elise; Dessailly, Florence; Oliver, Gérald; Martin, Florence; Montoro, Pascal

    2011-10-01

    Hevea brasiliensis transgenic plants are regenerated from transgenic callus lines by somatic embryogenesis. Somatic embryogenesis is not yet available for commercial propagation of Hevea clones, which requires conventional grafting of buds on rootstock seedlings (budding). The stability of transgene expression in budded plants is therefore necessary for further development of genetic engineering in rubber trees. Transgene expression was assessed by fluorimetric beta-glucuronidase (GUS) activity in fully developed leaves of in vitro plants from transgenic lines and their sub-lines obtained by budding. A large variation in GUS activity was found in self-rooted in vitro plants of five transgenic lines, and the absence of activity in one line suggested transgene silencing. Beyond confirming transmissibility of the reporter gene by budding and long-term expression, a quantification of GUS activity revealed that greater variability existed in budded plants compared to self-rooted mother in vitro plants for three transgenic lines. Although somatic embryogenesis provided more stable GUS activity, budding remained an efficient way of propagating transgenic plants but transgene expression in budded plants should be verified for functional analysis and further development. PMID:21643815

  3. [Coupling effects of water and chemical fertilizers on Hevea brasiliensis latex yield].

    PubMed

    Hua, Yuan-Gang; Chen, Qiu-Bo; Lin, Zhao-Mu; Luo, Wei

    2008-06-01

    Water and nutrient are the two main factors limiting Hevea brasiliensis growth and its latex yield. With 17 year-old Clone SCATC 7-33-97 H. brasiliensis as test material, the coupling effects of water and chemical N, P and K fertilizers on latex yield were studied by general orthogonal rotation design of quadratic regression with four factors and five levels under field condition, and a regressive mathematical model was set up based on the latex yield by quadratic regression analysis. The results showed that all test coupling levels of water and chemical fertilizers had significant effects on the latex yield. The yield-increasing effect of test factors was in the order of N application rate > irrigation amount > P application rate > K application rate, while the coupling effect of water and chemical fertilizers was in the sequence of water and N > N and P > water and P > water and K. There was a negative coupling effect of K application rate and soil moisture content. For latex yield, the optimum application rates of chemical fertilizers were 476.39 kg x hm(-2) of urea, 187.70 kg x hm(-2) of superphosphate and 225.77 kg x hm(-2) of potassium chloride, and the optimum irrigation amount was to have 82.78% soil relative water content. PMID:18808010

  4. Isolation and molecular characterization of 1-aminocyclopropane-1-carboxylic acid synthase genes in Hevea brasiliensis.

    PubMed

    Zhu, Jia-Hong; Xu, Jing; Chang, Wen-Jun; Zhang, Zhi-Li

    2015-01-01

    Ethylene is an important factor that stimulates Hevea brasiliensis to produce natural rubber. 1-Aminocyclopropane-1-carboxylic acid synthase (ACS) is a rate-limiting enzyme in ethylene biosynthesis. However, knowledge of the ACS gene family of H. brasiliensis is limited. In this study, nine ACS-like genes were identified in H. brasiliensis. Sequence and phylogenetic analysis results confirmed that seven isozymes (HbACS1-7) of these nine ACS-like genes were similar to ACS isozymes with ACS activity in other plants. Expression analysis results showed that seven ACS genes were differentially expressed in roots, barks, flowers, and leaves of H. brasiliensis. However, no or low ACS gene expression was detected in the latex of H. brasiliensis. Moreover, seven genes were differentially up-regulated by ethylene treatment. These results provided relevant information to help determine the functions of the ACS gene in H. brasiliensis, particularly the functions in regulating ethylene stimulation of latex production. PMID:25690030

  5. Cloning and Expression Analysis of One Gamma-Glutamylcysteine Synthetase Gene (Hbγ-ECS1) in Latex Production in Hevea brasiliensis

    PubMed Central

    Fang, Wei; Qiao, Luo Shi; Ming, Wu; Jian, Qiu; Feng, Yang Wen; Hua, Gao Hong; Zhou, Xiao Xian

    2016-01-01

    Rubber tree is a major commercial source of natural rubber. Latex coagulation is delayed by thiols, which belong to the important type of antioxidants in laticifer submembrane, and is composed of glutathione (GSH), cysteine, and methionine. The rate-limiting enzyme, γ-ECS, plays an important role in regulating the biosynthesis of glutathione under any environment conditions. To understand the relation between γ-ECS and thiols and to correlate latex flow with one-time tapping and continuous tapping, we cloned and derived the full length of one γ-ECS from rubber tree latex (Hbγ-ECS1). According to qPCR analysis, the expression levels of Hbγ-ECS1 were induced by tapping and Ethrel stimulation, and the expression was related to thiols content in the latex. Continuous tapping induced injury, and the expression of HbγECS1 increased with routine tapping and Ethrel-stimulation tapping (more intensive tapping). According to expression in long-term flowing latex, the gene was related to the duration of latex flow. HbγECS1 was expressed in E. coli Rosetta using pET-sumo as an expression vector and the recombinant enzyme was purified; then we achieved 0.827 U/mg specific activity and about 66 kDa molecular weight. The present study can help us understand the complex role of Hbγ-ECS in thiols biosynthesis, which is influenced by tapping. PMID:27419133

  6. Cloning and Expression Analysis of One Gamma-Glutamylcysteine Synthetase Gene (Hbγ-ECS1) in Latex Production in Hevea brasiliensis.

    PubMed

    Fang, Wei; Qiao, Luo Shi; Ming, Wu; Jian, Qiu; Feng, Yang Wen; Hua, Gao Hong; Zhou, Xiao Xian

    2016-01-01

    Rubber tree is a major commercial source of natural rubber. Latex coagulation is delayed by thiols, which belong to the important type of antioxidants in laticifer submembrane, and is composed of glutathione (GSH), cysteine, and methionine. The rate-limiting enzyme, γ-ECS, plays an important role in regulating the biosynthesis of glutathione under any environment conditions. To understand the relation between γ-ECS and thiols and to correlate latex flow with one-time tapping and continuous tapping, we cloned and derived the full length of one γ-ECS from rubber tree latex (Hbγ-ECS1). According to qPCR analysis, the expression levels of Hbγ-ECS1 were induced by tapping and Ethrel stimulation, and the expression was related to thiols content in the latex. Continuous tapping induced injury, and the expression of HbγECS1 increased with routine tapping and Ethrel-stimulation tapping (more intensive tapping). According to expression in long-term flowing latex, the gene was related to the duration of latex flow. HbγECS1 was expressed in E. coli Rosetta using pET-sumo as an expression vector and the recombinant enzyme was purified; then we achieved 0.827 U/mg specific activity and about 66 kDa molecular weight. The present study can help us understand the complex role of Hbγ-ECS in thiols biosynthesis, which is influenced by tapping. PMID:27419133

  7. Relationship between latex yield of Hevea brasiliensis and antecedent environmental parameters

    NASA Astrophysics Data System (ADS)

    Raj, Shammi; Das, Gitali; Pothen, Jacob; Dey, Sushil Kumar

    2005-01-01

    A study on the relationship between latex yield and antecedent environmental data was undertaken for five clones (RRII203, RRII118, RRIM600, RRII105 and GT1) of Hevea brasiliensis (rubber) in Agartala, northeast India, a region in which rubber is not traditionally cultivated. The explained variance for the regression equations based on parameters determined on the day of tapping and up to 3 days prior to it, varied from 72% to 37% during the NWT period and 94 83% during the WT period. Soil moisture storage, 1 and 3 days prior to tapping, was found to be the primary parameter affecting yield for the NWT and WT periods, respectively. It was observed that the clone RRII105, with a comparatively lower yield to that of RRIM600, was more susceptible to daily WD conditions during the non-winter season. RRIM600 and RRII105 being high-yielding clones were also found to be fairly dependent on the AT of the day prior to tapping. The mean lag period correlation of this parameter with yield was also found to be higher during the WT period than during the NWT period. As a whole, the mean lag period based on prior measurements of environmental variables showed optimum correlation with yield at 15 20 days prior to the day of tapping. The study also confirms that varied responses of yield with environmental factors in this non-traditional region of rubber cultivation depend on clonal character.

  8. Sucrose importation into laticifers of Hevea brasiliensis, in relation to ethylene stimulation of latex production

    PubMed Central

    Dusotoit-Coucaud, Anaïs; Brunel, Nicole; Kongsawadworakul, Panida; Viboonjun, Unchera; Lacointe, André; Julien, Jean-Louis; Chrestin, Hervé; Sakr, Soulaïman

    2009-01-01

    Background and Aims The major economic product of Hevea brasiliensis is a rubber-containing cytoplasm (latex), which flows out of laticifers (latex cells) when the bark is tapped. The latex yield is stimulated by ethylene. Sucrose, the unique precursor of rubber synthesis, must cross the plasma membrane through specific sucrose transporters before being metabolized in the laticifers. The relative importance of sucrose transporters in determining latex yield is unknown. Here, the effects of ethylene (by application of Ethrel®) on sucrose transporter gene expression in the inner bark tissues and latex cells of H. brasiliensis are described. Methods Experiments, including cloning sucrose transporters, real time RT-PCR and in situ hybridization, were carried out on virgin (untapped) trees, treated or untreated with the latex yield stimulant Ethrel. Key Results Seven putative full-length cDNAs of sucrose transporters were cloned from a latex-specific cDNA library. These transporters belong to all SUT (sucrose transporter) groups and differ by their basal gene expression in latex and inner soft bark, with a predominance of HbSUT1A and HbSUT1B. Of these sucrose transporters, only HbSUT1A and HbSUT2A were distinctly increased by ethylene. Moreover, this increase was shown to be specific to laticifers and to ethylene application. Conclusion The data and all previous information on sucrose transport show that HbSUT1A and HbSUT2A are related to the increase in sucrose import into laticifers, required for the stimulation of latex yield by ethylene in virgin trees. PMID:19567416

  9. Hydroxynitrile lyase from Hevea brasiliensis: molecular characterization and mechanism of enzyme catalysis.

    PubMed

    Hasslacher, M; Kratky, C; Griengl, H; Schwab, H; Kohlwein, S D

    1997-03-01

    (S)-Hydroxynitrile lyase (Hnl) from the tropical rubber tree Hevea brasiliensis is a 29 kDa single chain protein that catalyses the breakdown or formation of a C--C bond by reversible addition of hydrocyanic acid to aldehydes or ketones. The primary sequence of Hnl has no significant homology to known proteins. Detailed homology investigations employing PROFILESEARCH and secondary structure prediction algorithms suggest that Hnl is a member of the alpha/beta hydrolase fold protein family and contains a catalytic triad as functional residues for catalysis. The significance of predicted catalytic residues was tested and confirmed by site-directed mutagenesis and expression of mutant and wild-type proteins in the yeast, Saccharomyces cerevisiae. Based on these data we suggest a mechanistic model for the (S)-cyanohydrin synthesis catalyzed by hydroxynitrile lyase from Hevea brasiliensis. PMID:9094745

  10. Identification of laticifer-specific genes and their promoter regions from a natural rubber producing plant Hevea brasiliensis.

    PubMed

    Aoki, Yuichi; Takahashi, Seiji; Takayama, Daisuke; Ogata, Yoshiyuki; Sakurai, Nozomu; Suzuki, Hideyuki; Asawatreratanakul, Kasem; Wititsuwannakul, Dhirayos; Wititsuwannakul, Rapepun; Shibata, Daisuke; Koyama, Tanetoshi; Nakayama, Toru

    2014-08-01

    Latex, the milky cytoplasm of highly differentiated cells called laticifers, from Hevea brasiliensis is a key source of commercial natural rubber production. One way to enhance natural rubber production would be to express genes involved in natural rubber biosynthesis by a laticifer-specific overexpression system. As a first step to identify promoters which could regulate the laticifer-specific expression, we identified random clones from a cDNA library of H. brasiliensis latex, resulting in 4325 expressed sequence tags (ESTs) assembled into 1308 unigenes (692 contigs and 617 singletons). Quantitative analyses of the transcription levels of high redundancy clones in the ESTs revealed genes highly and predominantly expressed in laticifers, such as Rubber Elongation Factor (REF), Small Rubber Particle Protein and putative protease inhibitor proteins. HRT1 and HRT2, cis-prenyltransferases involved in rubber biosynthesis, was also expressed predominantly in laticifers, although these transcript levels were 80-fold lower than that of REF. The 5'-upstream regions of these laticifer-specific genes were cloned and analyzed in silico, revealing seven common motifs consisting of eight bases. Furthermore, transcription factors specifically expressed in laticifers were also identified. The common motifs in the laticifer-specific genes and the laticifer-specific transcription factors are potentially involved in the regulation of gene expression in laticifers. PMID:25017153

  11. Development, characterization, and cross-species/genera transferability of SSR markers for rubber tree (Hevea brasiliensis).

    PubMed

    Yu, Fei; Wang, Bao-Hua; Feng, Su-Ping; Wang, Jing-Yi; Li, Wei-Guo; Wu, Yao-Ting

    2011-03-01

    Genomic simple sequence repeat (SSR) markers are particularly valuable in studies of genetic diversity, evolution, genetic linkage map construction, quantitative trait loci tagging, and marker-assisted selection because of their multi-allelic nature, reproducibility, co-dominant inheritance, high abundance, and extensive genome coverage. The traditional methods of SSR marker development, such as genomic-SSR hybrid screening and microsatellite enrichment, have the disadvantages of high cost and complex operation. The selectively amplified microsatellite method is less costly and highly efficient as well as being simple and convenient. In this study, 252 sequences with SSRs were cloned from the rubber tree (Hevea brasiliensis) genome from which 258 SSR loci were obtained. The average repeat number was six. There were only 10 (3.9%) mononucleotide, trinucleotide, and pentanucleotide repeats, whereas the remaining 248 (96.1%) were dinucleotide repeats, including 128 (49.6%) GT/CA repeats, 118 (45.7%) GA/CT repeats, and 2 (0.8%) AT/TA repeats. A total of 126 primer pairs (see ESM) were successfully designed of which 36 primer pairs generated polymorphic products from 12 accessions of the cultivated species, 4 related species, and 3 species of the family Euphorbiaceae. In addition, investigations based on four genomic SSRs (GAR4, ACR22, CTR25, and GTR28) by cloning and sequencing provided evidence for cross-species/genera applicability, and homologous sequences were obtained from the rubber tree and Euphorbiaceae. Further analysis about the variation of the flanking regions of the four markers was carried out. PMID:20960206

  12. Sequence and Expression Analyses of Ethylene Response Factors Highly Expressed in Latex Cells from Hevea brasiliensis

    PubMed Central

    Piyatrakul, Piyanuch; Yang, Meng; Putranto, Riza-Arief; Pirrello, Julien; Dessailly, Florence; Hu, Songnian; Summo, Marilyne; Theeravatanasuk, Kannikar; Leclercq, Julie; Kuswanhadi; Montoro, Pascal

    2014-01-01

    The AP2/ERF superfamily encodes transcription factors that play a key role in plant development and responses to abiotic and biotic stress. In Hevea brasiliensis, ERF genes have been identified by RNA sequencing. This study set out to validate the number of HbERF genes, and identify ERF genes involved in the regulation of latex cell metabolism. A comprehensive Hevea transcriptome was improved using additional RNA reads from reproductive tissues. Newly assembled contigs were annotated in the Gene Ontology database and were assigned to 3 main categories. The AP2/ERF superfamily is the third most represented compared with other transcription factor families. A comparison with genomic scaffolds led to an estimation of 114 AP2/ERF genes and 1 soloist in Hevea brasiliensis. Based on a phylogenetic analysis, functions were predicted for 26 HbERF genes. A relative transcript abundance analysis was performed by real-time RT-PCR in various tissues. Transcripts of ERFs from group I and VIII were very abundant in all tissues while those of group VII were highly accumulated in latex cells. Seven of the thirty-five ERF expression marker genes were highly expressed in latex. Subcellular localization and transactivation analyses suggested that HbERF-VII candidate genes encoded functional transcription factors. PMID:24971876

  13. Enzymatic synthesis of rubber polymer in Hevea brasiliensis

    SciTech Connect

    Tang, F.; Hu, S.; Benedict, C.R. )

    1991-05-01

    Light and Dennis purified serum soluble rubber transferase from Hevea latex to homogeneity. Prenyl transferase co-purified with rubber transferase. In the absence of washed rubber particles (WRP) the prenyl transferase catalyzed the formation of trans FPP from DMAPP and IPP. In the presence of WRP the transferase catalyzed cis additions of IPP to pre-existing rubber chains. Control mixtures of WRP, Mg{sup 2+} and FPP were not included to test for the contributions of the bound rubber transferase on WRP to the incorporation of IPP into polyisoprene. Bound rubber transferase catalyzes the repetitive addition of IPP to allylic-PP starter molecules to form polyisoprene. The order of utilization of allylic-PP starters was GGPP > FPP > GPP > DMAPP. The authors have shown that the polyisoprene enzymatically synthesized on WRP is a bimodal polymer consisting of different mol wt rubber chains similar to the polymeric characteristics of natural rubber. The bound rubber transferase was solubilized with Chaps and purified on DEAE-cellulose. The polymerization reaction catalyzed by the purified preparation showed a 98% requirement for pre-existing rubber chains. Results suggest that the prenyl transferase from Hevea serum may be part of the polymer starter system furnishing allylic-PP for the bound rubber transferase.

  14. RNA sequencing read depth requirement for optimal transcriptome coverage in Hevea brasiliensis

    PubMed Central

    2014-01-01

    Background One of the concerns of assembling de novo transcriptomes is determining the amount of read sequences required to ensure a comprehensive coverage of genes expressed in a particular sample. In this report, we describe the use of Illumina paired-end RNA-Seq (PE RNA-Seq) reads from Hevea brasiliensis (rubber tree) bark to devise a transcript mapping approach for the estimation of the read amount needed for deep transcriptome coverage. Findings We optimized the assembly of a Hevea bark transcriptome based on 16 Gb Illumina PE RNA-Seq reads using the Oases assembler across a range of k-mer sizes. We then assessed assembly quality based on transcript N50 length and transcript mapping statistics in relation to (a) known Hevea cDNAs with complete open reading frames, (b) a set of core eukaryotic genes and (c) Hevea genome scaffolds. This was followed by a systematic transcript mapping process where sub-assemblies from a series of incremental amounts of bark transcripts were aligned to transcripts from the entire bark transcriptome assembly. The exercise served to relate read amounts to the degree of transcript mapping level, the latter being an indicator of the coverage of gene transcripts expressed in the sample. As read amounts or datasize increased toward 16 Gb, the number of transcripts mapped to the entire bark assembly approached saturation. A colour matrix was subsequently generated to illustrate sequencing depth requirement in relation to the degree of coverage of total sample transcripts. Conclusions We devised a procedure, the “transcript mapping saturation test”, to estimate the amount of RNA-Seq reads needed for deep coverage of transcriptomes. For Hevea de novo assembly, we propose generating between 5–8 Gb reads, whereby around 90% transcript coverage could be achieved with optimized k-mers and transcript N50 length. The principle behind this methodology may also be applied to other non-model plants, or with reads from other second generation

  15. Climate and natural production of rubber ( Hevea brasiliensis) in Xishuangbanna, southern part of Yunnan province, China

    NASA Astrophysics Data System (ADS)

    Jiang, Ailiang

    1988-12-01

    According to the author's and his collaborators' investigations, the climate influences the growth of rubber trees ( Hevea brasiliensis) in Xishuangbanna, the southern part of Yunnan Province, China, in at least four aspects: (1) The yield of latex per tapping and the final yield of dry rubber per tree per year or per unit area per year; (2) the growth rate, as expressed by increment of girth in cm; (3) the survival during the over-wintering period; (4) the initiation or suppression of certain diseases; In this paper the author would like to describe the influence of climatic elements on yield of latex and on survival during the over-wintering period. As for the other two aspects, only general comments are given.

  16. Synchronous flowering of the rubber tree (Hevea brasiliensis) induced by high solar radiation intensity.

    PubMed

    Yeang, Hoong-Yeet

    2007-01-01

    How tropical trees flower synchronously near the equator in the absence of significant day length variation or other meteorological cues has long been a puzzle. The rubber tree (Hevea brasiliensis) is used as a model to investigate this phenomenon. The annual cycle of solar radiation intensity is shown to correspond closely with the flowering of the rubber tree planted near the equator and in the subtropics. Unlike in temperate regions, where incoming solar radiation (insolation) is dependent on both day length and radiation intensity, insolation at the equator is due entirely to the latter. Insolation at the upper atmosphere peaks twice a year during the spring and autumn equinoxes, but the actual solar radiation that reaches the ground is attenuated to varying extents in different localities. The rubber tree shows one or two flowering seasons a year (with major and minor seasons in the latter) in accordance with the solar radiation intensity received. High solar radiation intensity, and in particular bright sunshine (as distinct from prolonged diffuse radiation), induces synchronous anthesis and blooming in Hevea around the time of the equinoxes. The same mechanism may be operational in other tropical tree species. PMID:17587376

  17. Identification and characterization of MAGO and Y14 genes in Hevea brasiliensis.

    PubMed

    Yang, Zi-Ping; Li, Hui-Liang; Guo, Dong; Peng, Shi-Qing

    2016-03-01

    Mago nashi (MAGO) and Y14 proteins are highly conserved among eukaryotes. In this study, we identified two MAGO (designated as HbMAGO1 andHbMAGO2) and two Y14 (designated as HbY14aand HbY14b) genes in the rubber tree (Hevea brasiliensis) genome annotation. Multiple amino acid sequence alignments predicted that HbMAGO and HbY14 proteins are structurally similar to homologous proteins from other species. Tissue-specific expression profiles showed that HbMAGO and HbY14 genes were expressed in at least one of the tissues (bark, flower, latex, leaf and root) examined. HbMAGOs and HbY14s were predominately located in the nucleus and were found to interact in yeast two-hybrid analysis (YTH) and bimolecular fluorescence complementation (BiFC) assays. HbMAGOs and HbY14s showed the highest transcription in latex and were regulated by ethylene and jasmonate. Interaction between HbMAGO2 and gp91phox (a large subunit of nicotinamide adenine dinucleotide phosphate) was identified using YTH and BiFC assays. These findings suggested that HbMAGO may be involved in the aggregation of rubber particles in H. brasiliensis. PMID:27007901

  18. Green synthesis of colloidal silver nanoparticles using natural rubber latex extracted from Hevea brasiliensis

    NASA Astrophysics Data System (ADS)

    Guidelli, Eder José; Ramos, Ana Paula; Zaniquelli, Maria Elisabete D.; Baffa, Oswaldo

    2011-11-01

    Colloidal silver nanoparticles were synthesized by an easy green method using thermal treatment of aqueous solutions of silver nitrate and natural rubber latex (NRL) extracted from Hevea brasiliensis. The UV-Vis spectra detected the characteristic surface plasmonic absorption band around 435 nm. Both NRL and AgNO 3 contents in the reaction medium have influence in the Ag nanoparticles formation. Lower AgNO 3 concentration led to decreased particle size. The silver nanoparticles presented diameters ranging from 2 nm to 100 nm and had spherical shape. The selected area electron diffraction (SAED) patterns indicated that the silver nanoparticles have face centered cubic (fcc) crystalline structure. FTIR spectra suggest that reduction of the silver ions are facilitated by their interaction with the amine groups from ammonia, which is used for conservation of the NRL, whereas the stability of the particles results from cis-isoprene binding onto the surface of nanoparticles. Therefore natural rubber latex extracted from H. brasiliensis can be employed in the preparation of stable aqueous dispersions of silver nanoparticles acting as a dispersing and/or capping agent. Moreover, this work provides a new method for the synthesis of silver nanoparticles that is simple, easy to perform, pollutant free and inexpensive.

  19. Identification and characterization of MAGO and Y14 genes in Hevea brasiliensis

    PubMed Central

    Yang, Zi-Ping; Li, Hui-Liang; Guo, Dong; Peng, Shi-Qing

    2016-01-01

    Abstract Mago nashi (MAGO) and Y14 proteins are highly conserved among eukaryotes. In this study, we identified two MAGO (designated as HbMAGO1 andHbMAGO2) and two Y14 (designated as HbY14aand HbY14b) genes in the rubber tree (Hevea brasiliensis) genome annotation. Multiple amino acid sequence alignments predicted that HbMAGO and HbY14 proteins are structurally similar to homologous proteins from other species. Tissue-specific expression profiles showed that HbMAGO and HbY14 genes were expressed in at least one of the tissues (bark, flower, latex, leaf and root) examined. HbMAGOs and HbY14s were predominately located in the nucleus and were found to interact in yeast two-hybrid analysis (YTH) and bimolecular fluorescence complementation (BiFC) assays. HbMAGOs and HbY14s showed the highest transcription in latex and were regulated by ethylene and jasmonate. Interaction between HbMAGO2 and gp91phox (a large subunit of nicotinamide adenine dinucleotide phosphate) was identified using YTH and BiFC assays. These findings suggested that HbMAGO may be involved in the aggregation of rubber particles in H. brasiliensis. PMID:27007901

  20. Hevea brasiliensis REF (Hev b 1) and SRPP (Hev b 3): An overview on rubber particle proteins.

    PubMed

    Berthelot, Karine; Lecomte, Sophie; Estevez, Yannick; Peruch, Frédéric

    2014-11-01

    This review article aims to gather all the knowledge on two important proteins associated with Hevea brasiliensis rubber particles: namely the rubber elongation factor (REF) and the small rubber particle protein (SRPP). It covers more then three decades of research on these two proteins and their homologues in plants, and particularly emphasizes on the different possible properties or functions of these various proteins found in plants. PMID:25019490

  1. Homologous Hevea brasiliensis REF (Hevb1) and SRPP (Hevb3) present different auto-assembling.

    PubMed

    Berthelot, Karine; Lecomte, Sophie; Estevez, Yannick; Coulary-Salin, Bénédicte; Peruch, Frédéric

    2014-02-01

    HbREF and HbSRPP are two Hevea brasiliensis proteins present on rubber particles, and probably involved in the coagulation of latex. Their function is unclear, but we previously discovered that REF had amyloid properties, which could be of particular interest during the coagulation process. First, we confirmed that REF and SRPP, homologous and principal proteins in hevea latex, are not glycoproteins. In this work, we investigated various aspects of protein interactions: aggregation, auto-assembling, yeast and erythrocyte agglutination, co-interactions by various biochemical (PAGE, spectroscopy, microscopy), biophysical (DLS, ellipsometry) and structural (TEM, ATR-FTIR, PM-IRRAS) approaches. We demonstrated that both proteins are auto-assembling into different aggregative states: REF polymerizes as an amyloid rich in β-sheets and forms quickly large aggregates (>μm), whereas SRPP auto-assembles in solution into stable nanomultimers of a more globular nature. Both proteins are however able to interact together, and SRPP may inhibit the amyloidogenesis of REF. REF is also able to interact with the membranes of yeasts and erythrocytes, leading to their agglutination. In addition, we also showed that both REF and SRPP did not have antimicrobial activity, whereas their activity on membranes has been clearly evidenced. We may suspect that these aggregative properties, even though they are clearly different, may occur during coagulation, when the membrane is destabilized. The interaction of proteins with membranes could help in the colloidal stability of latex, whereas the protein-protein interactions would contribute to the coagulation process, by bringing rubber particles together or eventually disrupting the particle monomembranes. PMID:24239687

  2. Ethylene Response Factors Are Controlled by Multiple Harvesting Stresses in Hevea brasiliensis.

    PubMed

    Putranto, Riza-Arief; Duan, Cuifang; Kuswanhadi; Chaidamsari, Tetty; Rio, Maryannick; Piyatrakul, Piyanuch; Herlinawati, Eva; Pirrello, Julien; Dessailly, Florence; Leclercq, Julie; Bonnot, François; Tang, Chaorong; Hu, Songnian; Montoro, Pascal

    2015-01-01

    Tolerance of recurrent mechanical wounding and exogenous ethylene is a feature of the rubber tree. Latex harvesting involves tapping of the tree bark and ethephon is applied to increase latex flow. Ethylene is an essential element in controlling latex production. The ethylene signalling pathway leads to the activation of Ethylene Response Factor (ERF) transcription factors. This family has been identified in Hevea brasiliensis. This study set out to understand the regulation of ERF genes during latex harvesting in relation to abiotic stress and hormonal treatments. Analyses of the relative transcript abundance were carried out for 35 HbERF genes in latex, in bark from mature trees and in leaves from juvenile plants under multiple abiotic stresses. Twenty-one HbERF genes were regulated by harvesting stress in laticifers, revealing an overrepresentation of genes in group IX. Transcripts of three HbERF-IX genes from HbERF-IXc4, HbERF-IXc5 and HbERF-IXc6 were dramatically accumulated by combining wounding, methyl jasmonate and ethylene treatments. When an ethylene inhibitor was used, the transcript accumulation for these three genes was halted, showing ethylene-dependent induction. Subcellular localization and transactivation experiments confirmed that several members of HbERF-IX are activator-type transcription factors. This study suggested that latex harvesting induces mechanisms developed for the response to abiotic stress. These mechanisms probably depend on various hormonal signalling pathways. Several members of HbERF-IX could be essential integrators of complex hormonal signalling pathways in Hevea. PMID:25906196

  3. PARTICLE SIZE INVESTIGATION ON NATURAL RUBBER FROM DIFFERENT CLONES OF BRAZILIAN IAC SERIES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Hevea brasiliensis species are being investigated in order to increase the production of natural rubber and to develop new clones more appropriate to specific soil and climate. Particle size of natural rubber from Hevea brasiliensis (Willd. ex Adr. de Juss.) Muell. - Arg. coming from new clones from...

  4. Hevea brasiliensis cell suspension peroxidase: purification, characterization and application for dye decolorization

    PubMed Central

    2013-01-01

    Peroxidases are oxidoreductase enzymes produced by most organisms. In this study, a peroxidase was purified from Hevea brasiliensis cell suspension by using anion exchange chromatography (DEAE-Sepharose), affinity chromatography (Con A-agarose) and preparative SDS-PAGE. The obtained enzyme appeared as a single band on SDS-PAGE with molecular mass of 70 kDa. Surprisingly, this purified peroxidase also had polyphenol oxidase activity. However, the biochemical characteristics were only studied in term of peroxidase because similar experiments in term of polyphenol oxidase have been reported in our pervious publication. The optimal pH of the purified peroxidase was 5.0 and its activity was retained at pH values between 5.0–10.0. The enzyme was heat stable over a wide range of temperatures (0–60°C), and less than 50% of its activity was lost at 70°C after incubation for 30 min. The enzyme was completely inhibited by β-mercaptoethanol and strongly inhibited by NaN3; in addition, its properties indicated that it was a heme containing glycoprotein. This peroxidase could decolorize many dyes; aniline blue, bromocresol purple, brilliant green, crystal violet, fuchsin, malachite green, methyl green, methyl violet and water blue. The stability against high temperature and extreme pH supported that the enzyme could be a potential peroxidase source for special industrial applications. PMID:23402438

  5. Diversity of the cassiicolin gene in Corynespora cassiicola and relation with the pathogenicity in Hevea brasiliensis.

    PubMed

    Déon, Marine; Fumanal, Boris; Gimenez, Stéphanie; Bieysse, Daniel; Oliveira, Ricardo R; Shuib, Siti Shuhada; Breton, Frédéric; Elumalai, Sunderasan; Vida, João B; Seguin, Marc; Leroy, Thierry; Roeckel-Drevet, Patricia; Pujade-Renaud, Valérie

    2014-01-01

    Corynespora cassiicola is an important plant pathogenic Ascomycete causing the damaging Corynespora Leaf Fall (CLF) disease in rubber tree (Hevea brasiliensis). A small secreted glycoprotein named cassiicolin was previously described as an important effector of C. cassiicola. In this study, the diversity of the cassiicolin-encoding gene was analysed in C. cassiicola isolates sampled from various hosts and geographical origins. A cassiicolin gene was detected in 47 % of the isolates, encoding up to six distinct protein isoforms. In three isolates, two gene variants encoding cassiicolin isoforms Cas2 and Cas6 were found in the same isolate. A phylogenetic tree based on four combined loci and elucidating the diversity of the whole collection was strongly structured by the toxin class, as defined by the cassiicolin isoform. The isolates carrying the Cas1 gene (toxin class Cas1), all grouped in the same highly supported clade, were found the most aggressive on two rubber tree cultivars. Some isolates in which no Cas gene was detected could nevertheless generate moderate symptoms, suggesting the existence of other yet uncharacterized effectors. This study provides a useful base for future studies of C. cassiicola population biology and epidemiological surveys in various host plants. PMID:24433675

  6. Antioxidative polyphenols from Nigerian mistletoe Loranthus micranthus (Linn.) parasitizing on Hevea brasiliensis.

    PubMed

    Agbo, Matthias Onyebuchi; Lai, Daowan; Okoye, Festus B C; Osadebe, Patience O; Proksch, Peter

    2013-04-01

    Two new phenolic glycosides, linamarin gallate (1) and walsuraside B (2), together with nine known compounds, catechin (3), epicatechin (4), epicatechin 3-O-gallate (5), epicatechin 3-O-(3-O-methyl)gallate (6), epicatechin 3-O-(3,5-O-dimethyl)gallate (7), epicatechin 3-O-(3,4,5-O-trimethyl)gallate (8), quercetin 3-O-β-d-glucopyranoside (9), rutin (10), and peltatoside (11), were isolated from the leafy twigs of Nigerian mistletoe Loranthus micranthus (Linn.) parasitic on Hevea brasiliensis. Compound 1 was characterized as an unusual cyanogenic glycoside, while compound 8 was isolated for the first time from a natural source. This is the first report of a cyanogenic glycoside from mistletoes. The structures of the new compounds were unambiguously elucidated by 1D ((1)H, (13)C), 2D NMR (COSY, HSQC, and HMBC) and by mass spectroscopy. The antioxidant activities of the isolated compounds (1-11) were evaluated using the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay. PMID:23422225

  7. Changes in the Chemical Composition and Decay Resistance of Thermally-Modified Hevea brasiliensis Wood.

    PubMed

    Severo, Elias Taylor Durgante; Calonego, Fred Willians; Sansígolo, Cláudio Angeli; Bond, Brian

    2016-01-01

    In this study the effect of thermal treatment on the equilibrium moisture content, chemical composition and biological resistance to decay fungi of juvenile and mature Hevea brasiliensis wood (rubber wood) was evaluated. Samples were taken from a 53-year-old rubber wood plantation located in Tabapuã, Sao Paulo, Brazil. The samples were thermally-modified at 180°C, 200°C and 220°C. Results indicate that the thermal modification caused: (1) a significant increase in the extractive content and proportional increase in the lignin content at 220°C; (2) a significant decrease in the equilibrium moisture content, holocelluloses, arabinose, galactose and xylose content, but no change in glucose content; and (3) a significant increase in wood decay resistance against both Pycnoporus sanguineus (L.) Murrill and Gloeophyllum trabeum (Pers.) Murrill decay fungi. The greatest decay resistance was achieved from treatment at 220°C which resulted in a change in wood decay resistance class from moderately resistant to resistant. Finally, this study also demonstrated that the influence of thermal treatment in mature wood was lower than in juvenile wood. PMID:26986200

  8. Changes in the Chemical Composition and Decay Resistance of Thermally-Modified Hevea brasiliensis Wood

    PubMed Central

    2016-01-01

    In this study the effect of thermal treatment on the equilibrium moisture content, chemical composition and biological resistance to decay fungi of juvenile and mature Hevea brasiliensis wood (rubber wood) was evaluated. Samples were taken from a 53-year-old rubber wood plantation located in Tabapuã, Sao Paulo, Brazil. The samples were thermally-modified at 180°C, 200°C and 220°C. Results indicate that the thermal modification caused: (1) a significant increase in the extractive content and proportional increase in the lignin content at 220°C; (2) a significant decrease in the equilibrium moisture content, holocelluloses, arabinose, galactose and xylose content, but no change in glucose content; and (3) a significant increase in wood decay resistance against both Pycnoporus sanguineus (L.) Murrill and Gloeophyllum trabeum (Pers.) Murrill decay fungi. The greatest decay resistance was achieved from treatment at 220°C which resulted in a change in wood decay resistance class from moderately resistant to resistant. Finally, this study also demonstrated that the influence of thermal treatment in mature wood was lower than in juvenile wood. PMID:26986200

  9. De novo Transcriptome Analysis Reveals Distinct Defense Mechanisms by Young and Mature Leaves of Hevea brasiliensis (Para Rubber Tree).

    PubMed

    Fang, Yongjun; Mei, Hailiang; Zhou, Binhui; Xiao, Xiaohu; Yang, Meng; Huang, Yacheng; Long, Xiangyu; Hu, Songnian; Tang, Chaorong

    2016-01-01

    Along with changes in morphology in the course of maturation, leaves of Hevea brasiliensis become more resistant to leaf diseases, including the South American Leaf Blight (SALB), a devastating fungal disease of this economically important tree species. To understand the underlying mechanisms of this defense, and to identify the candidate genes involved, we sequenced the Hevea leaf transcriptome at four developmental stages (I to IV) by Illumina sequencing. A total of 62.6 million high-quality reads were generated, and assembled into 98,796 unique transcripts. We identified 3,905 differentially expressed genes implicated in leaf development, 67.8% (2,651) of which were during the transition to leaf maturation. The genes involved in cyanogenic metabolism, lignin and anthocyanin biosynthesis were noteworthy for their distinct patterns of expression between developing leaves (stages I to III) and mature leaves (stage IV), and the correlation with the change in resistance to SALB and the Oidium/Colletotrichum leaf fall. The results provide a first profile of the molecular events that relate to the dynamics of leaf morphology and defense strategies during Hevea leaf development. This dataset is beneficial to devising strategies to engineer resistance to leaf diseases as well as other in-depth studies in Hevea tree. PMID:27619402

  10. Identification and Characterization of the Glucose-6-Phosphate Dehydrogenase Gene Family in the Para Rubber Tree, Hevea brasiliensis

    PubMed Central

    Long, Xiangyu; He, Bin; Fang, Yongjun; Tang, Chaorong

    2016-01-01

    As a key enzyme in the pentose phosphate pathway (PPP), glucose-6-phosphate dehydrogenase (G6PDH) provides nicotinamide adenine dinucleotide phosphate (NADPH) and intermediary metabolites for rubber biosynthesis, and plays an important role in plant development and stress responses. In this study, four Hevea brasiliensis (Para rubber tree) G6PDH genes (HbG6PDH1 to 4) were identified and cloned using a genome-wide scanning approach. All four HbG6PDH genes encode functional G6PDH enzymes as shown by heterologous expression in E. coli. Phylogeny analysis and subcellular localization prediction show that HbG6PDH3 is a cytosolic isoform, while the other three genes (HbG6PDH1, 2 and 4) are plastidic isoforms. The subcellular locations of HbG6PDH3 and 4, two latex-abundant isoforms were further verified by transient expression in rice protoplasts. Enzyme activity assay and expression analysis showed HbG6PDH3 and 4 were implicated in PPP during latex regeneration, and to influence rubber production positively in rubber tree. The cytosolic HbG6PDH3 is a predominant isoform in latex, implying a principal role for this isoform in controlling carbon flow and NADPH production in the PPP during latex regeneration. The expression pattern of plastidic HbG6PDH4 correlates well with the degree of tapping panel dryness, a physiological disorder that stops the flow of latex from affected rubber trees. In addition, the four HbG6PDHs responded to temperature and drought stresses in root, bark, and leaves, implicating their roles in maintaining redox balance and defending against oxidative stress. PMID:26941770

  11. Identification and Characterization of the Glucose-6-Phosphate Dehydrogenase Gene Family in the Para Rubber Tree, Hevea brasiliensis.

    PubMed

    Long, Xiangyu; He, Bin; Fang, Yongjun; Tang, Chaorong

    2016-01-01

    As a key enzyme in the pentose phosphate pathway (PPP), glucose-6-phosphate dehydrogenase (G6PDH) provides nicotinamide adenine dinucleotide phosphate (NADPH) and intermediary metabolites for rubber biosynthesis, and plays an important role in plant development and stress responses. In this study, four Hevea brasiliensis (Para rubber tree) G6PDH genes (HbG6PDH1 to 4) were identified and cloned using a genome-wide scanning approach. All four HbG6PDH genes encode functional G6PDH enzymes as shown by heterologous expression in E. coli. Phylogeny analysis and subcellular localization prediction show that HbG6PDH3 is a cytosolic isoform, while the other three genes (HbG6PDH1, 2 and 4) are plastidic isoforms. The subcellular locations of HbG6PDH3 and 4, two latex-abundant isoforms were further verified by transient expression in rice protoplasts. Enzyme activity assay and expression analysis showed HbG6PDH3 and 4 were implicated in PPP during latex regeneration, and to influence rubber production positively in rubber tree. The cytosolic HbG6PDH3 is a predominant isoform in latex, implying a principal role for this isoform in controlling carbon flow and NADPH production in the PPP during latex regeneration. The expression pattern of plastidic HbG6PDH4 correlates well with the degree of tapping panel dryness, a physiological disorder that stops the flow of latex from affected rubber trees. In addition, the four HbG6PDHs responded to temperature and drought stresses in root, bark, and leaves, implicating their roles in maintaining redox balance and defending against oxidative stress. PMID:26941770

  12. HbNIN2, a cytosolic alkaline/neutral-invertase, is responsible for sucrose catabolism in rubber-producing laticifers of Hevea brasiliensis (para rubber tree).

    PubMed

    Liu, Shujin; Lan, Jixian; Zhou, Binhui; Qin, Yunxia; Zhou, Yihua; Xiao, Xiaohu; Yang, Jianghua; Gou, Jiqing; Qi, Jiyan; Huang, Yacheng; Tang, Chaorong

    2015-04-01

    In Hevea brasiliensis, an alkaline/neutral invertase (A/N-Inv) is responsible for sucrose catabolism in latex (essentially the cytoplasm of rubber-producing laticifers, the source of natural rubber) and implicated in rubber yield. However, neither the gene encoding this enzyme nor its molecular and biochemical properties have been well documented. Three Hevea A/N-Inv genes, namely HbNIN1, 2 and 3, were first cloned and characterized in planta and in Escherichia coli. Cellular localizations of HbNIN2 mRNA and protein were probed. From latex, active A/N-Inv proteins were purified, identified, and explored for enzymatic properties. HbNIN2 was identified as the major A/N-Inv gene functioning in latex based on its functionality in E. coli, its latex-predominant expression, the conspicuous localization of its mRNA and protein in the laticifers, and its expressional correlation with rubber yield. An active A/N-Inv protein was partially purified from latex, and determined as HbNIN2. The enhancement of HbNIN2 enzymatic activity by pyridoxal is peculiar to A/N-Invs in other plants. We conclude that HbNIN2, a cytosolic A/N-Inv, is responsible for sucrose catabolism in rubber laticifers. The results contribute to the studies of sucrose catabolism in plants as a whole and natural rubber synthesis in particular. PMID:25581169

  13. Isolation and structure elucidaton of polyphenols from Loranthus micranthus Linn. parasitic on Hevea brasiliensis with antiinflammatory property

    PubMed Central

    Agbo, Matthias Onyebuchi; Nworu, Chukwuemeka Sylvester; Okoye, Festus Basden Chied; Osadebe, Patience Ogoamaka

    2014-01-01

    The present study was carried out to evaluate the anti-inflammatory activities of polyphenols isolated from the leaves of mistletoe (Loranthus micranthus Linn.) parasitic on Hevea brasiliensis. The anti-inflammatory properties of the isolated compounds were evaluated on the basis of their ability to inhibit the production of nitric oxide (NO) and tumuor necrosis factor-α (TNF-α) in lipopolysaccharide (LPS) activated RAW 264.7 mouse macrophages. Semi-preparative HPLC separation of the ethyl acetate (EtOAc) and butanol (n-BuOH) fractions of the leaves of mistletoe (Loranthus micranthus Linn) parasitic on Hevea brasiliensis led to the isolation of four polyphenols: 3-O-(3,4,5-trimethoxybenzoyl)-(-)-epicatechin (TMECG) (1); (-)-epicatechin-3-O-(3″-O-methyl)-gallate (ECG3″Me) (2); rutin (3) and peltatoside (4). Compounds 1-4 were isolated for the first time from this plant while 1 was isolated for the first time in nature. These compounds (1-4) were readily identified by comparison of their spectroscopic data with those reported in the literature. The polyphenols proved to have anti-inflammatory activity as evidenced by the suppression of inducible nitric oxide (iNO) and cytokine (TNF-α) levels in the culture supernatant of lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages. However, the study showed that the quercetin diglycosides showed stronger inhibition of proinflammatory mediators than the epicatechin derivates. These data provide evidence that polyphenolic compounds isolated from the mistletoe parasitic on Hevea brasiliensis may contribute to its anti-inflammatory properties by inhibiting the expression of inducible nitric oxide and proinflammatory cytokines such as tumour necrosis factor-α. PMID:26417309

  14. Base temperature and heat units for leaf flushing emission and growth of Hevea brasiliensis Muell. Arg.

    NASA Astrophysics Data System (ADS)

    de Lemos Filho, José Pires; Villa Nova, Nilson Augusto; Silveira Pinto, Hilton

    1993-06-01

    In this study we determined the base temperature and Heat units for leaf flushing initiation and growth of RRIM-600 and GT1 Hevea clones. The minimum base temperature was found to be approximately 16° C for leaf flushing emission and 19° C for shoot growth in terms of height. Approximately 420 degree days at 16° C as base temperature is required for successive leaf flushing initiation. Linear equations with correlation coefficients above 0.95 allow an estimation of the height increase from the accumulated degree days, corrected or otherwise for photoperiod. The highest correlation coefficients demonstrated a positive effect of the photoperiod among the factors influencing the shoot growth.

  15. De novo transcriptome analysis of Hevea brasiliensis tissues by RNA-seq and screening for molecular markers

    PubMed Central

    2014-01-01

    Background The rubber tree, Hevea brasiliensis, is a species native to the Brazilian Amazon region and it supplies almost all the world’s natural rubber, a strategic raw material for a variety of products. One of the major challenges for developing rubber tree plantations is adapting the plant to biotic and abiotic stress. Transcriptome analysis is one of the main approaches for identifying the complete set of active genes in a cell or tissue for a specific developmental stage or physiological condition. Results Here, we report on the sequencing, assembling, annotation and screening for molecular markers from a pool of H. brasiliensis tissues. A total of 17,166 contigs were successfully annotated. Then, 2,191 Single Nucleotide Variation (SNV) and 1.397 Simple Sequence Repeat (SSR) loci were discriminated from the sequences. From 306 putative, mainly non-synonymous SNVs located in CDS sequences, 191 were checked for their ability to characterize 23 Hevea genotypes by an allele-specific amplification technology. For 172 (90%), the nucleotide variation at the predicted genomic location was confirmed, thus validating the different steps from sequencing to the in silico detection of the SNVs. Conclusions This is the first study of the H. brasiliensis transcriptome, covering a wide range of tissues and organs, leading to the production of the first developed SNP markers. This process could be amplified to a larger set of in silico detected SNVs in expressed genes in order to increase the marker density in available and future genetic maps. The results obtained in this study will contribute to the H. brasiliensis genetic breeding program focused on improving of disease resistance and latex yield. PMID:24670056

  16. Genome-wide identification and expression analysis of the metacaspase gene family in Hevea brasiliensis.

    PubMed

    Liu, Hui; Deng, Zhi; Chen, Jiangshu; Wang, Sen; Hao, Lili; Li, Dejun

    2016-08-01

    Metacaspases, a family of cysteine proteases, have been suggested to play important roles in programmed cell death (PCD) during plant development and stress responses. To date, no systematic characterization of this gene family has been reported in rubber tree (Hevea brasiliensis). In the present study, nine metacaspase genes, designated as HbMC1 to HbMC9, were identified from whole-genome sequence of rubber tree. Multiple sequence alignment and phylogenetic analyses suggested that these genes were divided into two types: type I (HbMC1-HBMC7) and type II (HbMC8 and HbMC9). Gene structure analysis demonstrated that type I and type II HbMCs separately contained four and two introns, indicating the conserved exon-intron organization of HbMCs. Quantitative real-time PCR analysis revealed that HbMCs showed distinct expression patterns in different tissues, suggesting the functional diversity of HbMCs in various tissues during development. Most of the HbMCs were regulated by drought, cold, and salt stress, implying their possible functions in regulating abiotic stress-induced cell death. Of the nine HbMCs, HbMC1, HbMC2, HbMC5, and HbMC8 displayed a significantly higher relative transcript accumulation in barks of tapping panel dryness (TPD) trees compared with healthy trees. In addition, the four genes were up-regulated by ethephon (ET) and methyl jasmonate (MeJA), indicating their potential involvement in TPD resulting from ET- or JA-induced PCD. In summary, this work provides valuable information for further functional characterization of HbMC genes in rubber tree. PMID:27085600

  17. Evapotranspiration of rubber (Hevea brasiliensis) cultivated at two plantation sites in Southeast Asia

    NASA Astrophysics Data System (ADS)

    Giambelluca, Thomas W.; Mudd, Ryan G.; Liu, Wen; Ziegler, Alan D.; Kobayashi, Nakako; Kumagai, Tomo'omi; Miyazawa, Yoshiyuki; Lim, Tiva Khan; Huang, Maoyi; Fox, Jefferson; Yin, Song; Mak, Sophea Veasna; Kasemsap, Poonpipope

    2016-02-01

    To investigate the effects of expanding rubber (Hevea brasiliensis) cultivation on water cycling in Mainland Southeast Asia (MSEA), evapotranspiration (ET) was measured within rubber plantations at Bueng Kan, Thailand, and Kampong Cham, Cambodia. After energy closure adjustment, mean annual rubber ET was 1211 and 1459 mm yr-1 at the Thailand and Cambodia sites, respectively, higher than that of other tree-dominated land covers in the region, including tropical seasonal forest (812-1140 mm yr-1), and savanna (538-1060 mm yr-1). The mean proportion of net radiation used for ET by rubber (0.725) is similar to that of tropical rainforest (0.729) and much higher than that of tropical seasonal forest (0.595) and savanna (0.548). Plant area index (varies with leaf area changes), explains 88.2% and 73.1% of the variance in the ratio of latent energy flux (energy equivalent of ET) to potential latent energy flux (LE/LEpot) for midday rain-free periods at the Thailand and Cambodia sites, respectively. High annual rubber ET results from high late dry season water use, associated with rapid refoliation by this brevideciduous species, facilitated by tapping of deep soil water, and by very high wet season ET, a characteristic of deciduous trees. Spatially, mean annual rubber ET increases strongly with increasing net radiation (Rn) across the three available rubber plantation observation sites, unlike nonrubber tropical ecosystems, which reduce canopy conductance at high Rn sites. High water use by rubber raises concerns about potential effects of continued expansion of tree plantations on water and food security in MSEA.

  18. Catalytic mechanism of hydroxynitrile lyase from Hevea brasiliensis: a theoretical investigation.

    PubMed

    Cui, Feng-Chao; Pan, Xiao-Liang; Liu, Jing-Yao

    2010-07-29

    Density functional theory (DFT) calculations using the hybrid functional B3LYP have been performed to investigate the catalytic mechanism of hydroxynitrile lyase from Hevea brasiliensis (Hb-HNL). This enzyme catalyzes the cleavage of acetone cyanohydrin to hydrocyanic acid plus acetone. Two models (A and B) of the active site consisting of 105 and 155 atoms, respectively, were constructed on the basis of the crystal structure. Good consistency between the two models provides a verification of the proposed mechanism. Our calculations show that the catalytic reaction proceeds via three elementary steps: (1) deprotonation of the OH-Ser80 by His235 and concomitant abstraction of a proton from the substrate hydroxyl by Ser80; (2) the C-C bond cleavage of the acetone cyanohydrin; and (3) protonation of the cleaved cyanide by His235. The cleavage of the C-C bond is the rate-limiting step with the overall free energy barrier of 13.5 kcal/mol for relatively smaller model A (14.9 kcal/mol for a larger model B) in the protein environment, which is in good agreement with experimental rate. The present results give support to the previously proposed general acid/base catalytic mechanism, in which the catalytic triad acts as a general acid/base. Moreover, the calculated results for model C, with the positive charge of Lys236 removed from model A, show that Lys236 with the positive charge plays a vital role in lowering the reaction barrier of the rate-determining and helps in stabilizing the negatively charged CN(-) by forming a hydrogen bond with the substrate, consistent with the experimental analysis. PMID:20593768

  19. Responses of seedlings of tropical woody plants to environmental stresses with emphasis on Theobroma cacao and Hevea brasiliensis

    SciTech Connect

    Sena Gomes, A.R.

    1987-01-01

    Relative humidity, flooding, temperature, wind, and SO/sub 2/ variously influenced physiological processes and growth of tropical woody plants, with emphasis on three Theobroma cacao varieties and three Hevea brasiliensis families. Stomata were smaller and more numerous in Theobroma than in Hevea. In Theobroma, but not Heavea, stomatal frequency decreased from the leaf base to the apex and from the midrib outward. Stomata of Theobroma cacao var. Catongo opened in high relative humidity (RH) and closed in low RH. The more open stomata in high RH were associated with high rates of photosynthesis, low leaf water potential, high water use efficiency (WUE), and low transpiration rate (TR). Variations in TR and WUE were correlated with changes in vapor pressure deficit. Other responses included stomatal closure, decreased chlorophyll content, leaf epinasty, production of hypertrophied lenticels and adventitious roots, and acceleration of ethylene production. Responses to flooding varied with species, Theobroma varieties and Hevea families. Effects of temperature regimes on growth varied with species, varieties and families, plant parts, growth parameters, and time of harvesting. Optimal temperatures for dry weight increase of stems or roots of Theobroma cacao var. Comum were 22.2 C; and 33.3 C for dry weight increase or relative growth rates of leaves or seedlings. Optimal temperatures for growth varied for Hevea families. Wind injured leaves of Theobroma cacao, with more injury by wind of 6.0 than 3.0 m s/sup -1/. Stomata were more open on windy than on calm days, but tended to close at high wind speeds. Wind lowered transpiration rate but the reduction was not correlated with leaf dehydration. SO/sub 2/ at 0.5, 1.0, or 1.5 ppm for 24 h did not injure Theobroma leaves but reduced dry weight increment of leaves of var. Catongo but not Catongo/Sial.

  20. How Do Rubber (Hevea brasiliensis) Plantations Cope with Seasonal Drought in Northern Thailand and Central Cambodia?

    NASA Astrophysics Data System (ADS)

    Kumagai, T.; Giambelluca, T. W.

    2014-12-01

    Rubber (Hevea brasiliensis Müll. Arg.) plantaitons are rapidly expanding throughout mainland Southeast Asia, potentially changing the partitioning of water, energy, and carbon at multiple scales, compared with the traditional land covers they are replacing. We have conducted eddy flux measurements in two rubber plantation sites: Som Sanuk (SS), located northern Thailand; and Cambodian Rubber Research Institute (CRRI), central Cambodia. We used combination of actual evapotranspiration (ET) flux measurements and an inversed version of a simple 2-layer ET model for estimating the mean canopy stomatal conductances (gs), which is among the most effective measures for describing the exchange characteristics. It is demonstrated how each studied rubber plantation copes with each strong seasonal drought via tree water use strategies. Potential tree water use deficit (precipitation (P) - potential evaporation (ET_POT)) for each season (i.e., December-February: DJF, March-May: MAM, June-August: JJA, and September-November: SON) revealed in which season and how the water use should be controlled. We found that in the season when actual tree water use deficit (P - ET) was negative (DJF and MAM), the deficit was compensated with soil water from the previous season at a depth of 0-2 m at the Thailand site, and from a depth of 0-3 m at CRRI. Two ecophysiological parameters, the reference value of gs (gsref) and the sensitivity of gs to atmospheric demand (m), as well as their proportionality (m/gsref), were derived from the logarithmic response curve of gs to vapor pressure deficit (D) for each season and each site. In both sites, gsref and m appeared to be less in DJF and MAM than each in the other three month periods (seasons). On average in a whole year, m/gsref was less than 0.6 at SS and almost 0.6 at the CRRI site, suggesting that there was less sufficient stomatal regulation at SS, where there might be little risk of water stress-induced hydraulic failure because of much

  1. De novo assembly and transcriptome analysis of the rubber tree (Hevea brasiliensis) and SNP markers development for rubber biosynthesis pathways.

    PubMed

    Mantello, Camila Campos; Cardoso-Silva, Claudio Benicio; da Silva, Carla Cristina; de Souza, Livia Moura; Scaloppi Junior, Erivaldo José; de Souza Gonçalves, Paulo; Vicentini, Renato; de Souza, Anete Pereira

    2014-01-01

    Hevea brasiliensis (Willd. Ex Adr. Juss.) Muell.-Arg. is the primary source of natural rubber that is native to the Amazon rainforest. The singular properties of natural rubber make it superior to and competitive with synthetic rubber for use in several applications. Here, we performed RNA sequencing (RNA-seq) of H. brasiliensis bark on the Illumina GAIIx platform, which generated 179,326,804 raw reads on the Illumina GAIIx platform. A total of 50,384 contigs that were over 400 bp in size were obtained and subjected to further analyses. A similarity search against the non-redundant (nr) protein database returned 32,018 (63%) positive BLASTx hits. The transcriptome analysis was annotated using the clusters of orthologous groups (COG), gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Pfam databases. A search for putative molecular marker was performed to identify simple sequence repeats (SSRs) and single nucleotide polymorphisms (SNPs). In total, 17,927 SSRs and 404,114 SNPs were detected. Finally, we selected sequences that were identified as belonging to the mevalonate (MVA) and 2-C-methyl-D-erythritol 4-phosphate (MEP) pathways, which are involved in rubber biosynthesis, to validate the SNP markers. A total of 78 SNPs were validated in 36 genotypes of H. brasiliensis. This new dataset represents a powerful information source for rubber tree bark genes and will be an important tool for the development of microsatellites and SNP markers for use in future genetic analyses such as genetic linkage mapping, quantitative trait loci identification, investigations of linkage disequilibrium and marker-assisted selection. PMID:25048025

  2. De Novo Assembly and Transcriptome Analysis of the Rubber Tree (Hevea brasiliensis) and SNP Markers Development for Rubber Biosynthesis Pathways

    PubMed Central

    Mantello, Camila Campos; Cardoso-Silva, Claudio Benicio; da Silva, Carla Cristina; de Souza, Livia Moura; Scaloppi Junior, Erivaldo José; de Souza Gonçalves, Paulo; Vicentini, Renato; de Souza, Anete Pereira

    2014-01-01

    Hevea brasiliensis (Willd. Ex Adr. Juss.) Muell.-Arg. is the primary source of natural rubber that is native to the Amazon rainforest. The singular properties of natural rubber make it superior to and competitive with synthetic rubber for use in several applications. Here, we performed RNA sequencing (RNA-seq) of H. brasiliensis bark on the Illumina GAIIx platform, which generated 179,326,804 raw reads on the Illumina GAIIx platform. A total of 50,384 contigs that were over 400 bp in size were obtained and subjected to further analyses. A similarity search against the non-redundant (nr) protein database returned 32,018 (63%) positive BLASTx hits. The transcriptome analysis was annotated using the clusters of orthologous groups (COG), gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Pfam databases. A search for putative molecular marker was performed to identify simple sequence repeats (SSRs) and single nucleotide polymorphisms (SNPs). In total, 17,927 SSRs and 404,114 SNPs were detected. Finally, we selected sequences that were identified as belonging to the mevalonate (MVA) and 2-C-methyl-D-erythritol 4-phosphate (MEP) pathways, which are involved in rubber biosynthesis, to validate the SNP markers. A total of 78 SNPs were validated in 36 genotypes of H. brasiliensis. This new dataset represents a powerful information source for rubber tree bark genes and will be an important tool for the development of microsatellites and SNP markers for use in future genetic analyses such as genetic linkage mapping, quantitative trait loci identification, investigations of linkage disequilibrium and marker-assisted selection. PMID:25048025

  3. Mites (Acari) of rubber trees (Hevea brasiliensis Muell. Arg., Euphorbiaceae) in Piracicaba, state of São Paulo, Brazil.

    PubMed

    De Vis, Raf M J; de Moraes, Gilberto J; Bellini, Marcos R

    2006-01-01

    This study determined the main mite species on rubber trees (clone RRIM-600) in Piracicaba, southeast of São Paulo State, from June 2002 to May 2003 and evaluated the possible relation between them. It was conducted in a plantation of 5 ha on 11 year old trees, 15 m high, surrounded with crops as pearl millet, cotton, bean or corn. Samples were taken monthly and consisted of five leaflets, five petioles (only from October 2002 on) and five terminal sections of twigs (10 cm) from 15 rubber trees. All mites of one leaflet, one petiole and one twig section of each plant were mounted for identification to genera/species to estimate the proportional occurrence of each species. A total of 84,850 mites belonging to 38 species of 34 genera and 16 families were found. Tydeidae was the family with the highest number of species (11), followed by Phytoseiidae and Stigmaeidae (4 each). The most abundant families were Eriophyidae, Tenuipalpidae and Tydeidae (totals of 43,023, 26,390 and 13,644 individuals, respectively). The highest population levels of the pest mites Calacarus heveae Feres and Tenuipalpus heveae Baker occurred at the end of the rainy season. The most abundant predators were Metaseiulus camelliae (Chant & Yoshida-Shaul), Amblyseius compositus Denmark & Muma and Euseius citrifolius Denmark & Muma. The predators could not prevent the increase of C. heveae and T. heveae from March on. However, their presence might have prevented an earlier increase and even higher levels of those mites. PMID:17352076

  4. Trichoderma amazonicum, a new endophytic species on Hevea brasiliensis and H. guianensis from the Amazon basin

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A new species of Trichoderma (teleomorph Hypocrea, Ascomycota, Sordariomycetes, Hypocreales, Hypocreaceae), T. amazonicum, endophytic on the living sapwood and leaves of Hevea spp. trees is described. Trichoderma amazonicum is distinguished from closely related species in the Harzianum clade (e.g. ...

  5. Molecular characterization and expression analysis of the small GTPase ROP members expressed in laticifers of the rubber tree (Hevea brasiliensis).

    PubMed

    Qin, Yunxia; Huang, Yacheng; Fang, Yongjun; Qi, Jiyan; Tang, Chaorong

    2014-01-01

    ROP (Rho of plants) proteins are plant-specific Rho-type small GTPases which play important roles in cellular processes and stress responses. This study explores the characteristics and possible functions of ROPs that are expressed primarily in laticifers of the rubber tree (Hevea brasiliensis). The work serves as a preliminary step to determining their involvement in latex flow and regeneration, laticifers formation and tapping panel dryness (TPD, a physiological disorder in rubber trees that result in the stoppage of latex flow). In this connection, we (i) identified five HbROPs (HbROP1-HbROP5) by searching latex transcripts database and the genome databases, (ii) characterized molecular and phylogenic aspects of the HbROPs and examined the cis-regulatory elements in their promoter regions; (iii) analyzed by Real-time Quantitative PCR (QPCR) the tissue specificity of the HbROPs and their expression patterns in response to tapping, bark wounding and growth regulator treatments. All five HbROP genes were strongly expressed in the latex, with HbROP1, 3, 4 and 5 showing the highest expression among the six Hevea tissues examined, viz. latex, bud, mature leaf, bark, male flower and seed. When tapping was initiated on previously untapped trees, HbROP3 transcription was substantially down-regulated whereas HbROP5 expression was markedly up-regulated. Transcripts of HbROP3 rose gradually with the development of TPD. Except for the cytokinin 6-benzyl aminopurine that induced a rise in HbROP5 transcripts by more than 2-fold, the other growth regulators tested had little effect on HbROPs expression. The roles of HbROPs in rubber tree are discussed in relation to the diverse functions of ROP homologs reported in other plant species. PMID:24308989

  6. Responses of Hevea brasiliensis Muell. Arg. plants to low temperatures under controlled conditions

    NASA Astrophysics Data System (ADS)

    de Lemos Filho, José Pires; Pinto, Hilton Silveira; da Rocha Neto, Olinto Gomes

    1993-12-01

    In this study we have examined the responses of Hevea plants to low temperatures in growth chambers simulating winter conditions in the tropics. The low temperatures increased the leaf diffusive resistance even with a rise in the leaf temperatures toward the end of day. The total chlorophyll levels were reduced by chilling, with no changes in the chlorophyll a/b ratio. Neither organic solute nor electrolyte leakage were observed at low temperatures. These results explain the low growth rate of plants and the lack of serious chilling symptoms (necrosis) in mature leaves during winter conditions near latitude 22° S in Brazil.

  7. Screening of bacterial biocontrols against sapstain fungus (Lasiodiplodia theobromae Pat.) of rubberwood (Hevea brasiliensis Muell.Arg.).

    PubMed

    Sajitha, K L; Maria Florence, E J; Dev, Suma Arun

    2014-09-01

    Diverse bacterial biocontrol agents from various sources of aerobic composts against the sapstain fungus Lasiodiplodia theobromae in rubberwood (Hevea brasiliensis) were isolated, screened and identified by various morphological, biochemical and molecular techniques. The inhibitory effect of seventeen bacterial isolates was examined and seven exhibited inhibition towards the sapstain fungus. Among the seven antagonists, six were conclusively identified as Bacillus subtilis and one as Paenibacillus polymyxa using 16S rRNA-encoding gene sequencing. This is the first report on the occurrence of P. polymyxa, a potent biofertilizer and antagonist in vermicompost. HiCrome Bacillus agar was identified as an effective medium for differentiation of B. subtilis from other Bacillus species. The present work demonstrates the efficacy of the antagonistic property of B. subtilis strains against rubberwood sapstain fungus. Culture-based antagonistic inhibition displayed by B. subtilis can be extended to cater to the biocontrol requirements of wood-based industries against the stain fungus. The study showed the utility of an integrated approach, employing morphological, biochemical and molecular tools for conclusive identification of several bacterial isolates present in aerobic composts from diverse sources. PMID:25049165

  8. Hevea brasiliensis prohevein possesses a conserved C-terminal domain with amyloid-like properties in vitro.

    PubMed

    Berthelot, Karine; Lecomte, Sophie; Coulary-Salin, Bénédicte; Bentaleb, Ahmed; Peruch, Frédéric

    2016-04-01

    Prohevein is a wound-induced protein and a main allergen from latex of Hevea brasiliensis (rubber tree). This 187 amino-acid protein is cleaved in two fragments: a N-terminal 43 amino-acids called hevein, a lectin bearing a chitin-binding motif with antifungal properties and a C-terminal domain (C-ter) far less characterized. We provide here new insights on the characteristics of prohevein, hevein and C-terminal domain. Using complementary biochemical (ThT/CR/chitin binding, agglutination) and structural (modeling, ATR-FTIR, TEM, WAXS) approaches, we show that this domain clearly displays all the characteristics of an amyloid-like proteins in vitro, that could confer agglutination activity in synergy with its chitin-binding activity. Additionally, this C-ter domain is highly conserved and present in numerous plant prohevein-like proteins or pathogenesis-related (PR and WIN) proteins. This could be the hallmark of the eventual presence of proteins with amyloid properties in plants, that could potentially play a role in defense through aggregation properties. PMID:26805576

  9. High-level intracellular expression of hydroxynitrile lyase from the tropical rubber tree Hevea brasiliensis in microbial hosts.

    PubMed

    Hasslacher, M; Schall, M; Hayn, M; Bona, R; Rumbold, K; Lückl, J; Griengl, H; Kohlwein, S D; Schwab, H

    1997-10-01

    (S)-Hydroxynitrile lyase (Hnl) from the tropical rubber tree Hevea brasiliensis catalyzes the formation of (S)-cyanohydrins from hydrocyanic acid and aldehydes or ketones. This enzyme accepts aliphatic, aromatic, and heterocyclic carbonyl compounds as substrates and is therefore considered a potent biocatalyst for the industrial production of optically active chemicals. Limitations in enzyme supply from natural resources were overcome by production of the enzyme in the microbial host systems Escherichia coli, Saccharomyces cerevisiae, and Pichia pastoris. Expression of Hnl in the prokaryotic system led to the formation of inclusion bodies whereas in both yeast hosts high levels of soluble protein were obtained. Highest yields were obtained in a high cell density batch fermentation of a P. pastoris transformant that expressed heterologous Hnl to about 50% of the soluble cytosolic protein. At a cell density of 100 g/liter cell dry weight, a volume yield of 22 g/liter of heterologous product was obtained. Attempts to produce the Hnl protein extracellularly with the yeast hosts by applying different leader peptide strategies were not successful. Immunofluorescence microscopy studies indicated that the secretion-directed heterologous Hnl protein accumulated in the plasma membrane forming aggregated clusters of inactive protein. PMID:9325140

  10. Comparative study of two GH19 chitinase-like proteins from Hevea brasiliensis, one exhibiting a novel carbohydrate-binding domain.

    PubMed

    Martínez-Caballero, Siseth; Cano-Sánchez, Patricia; Mares-Mejía, Israel; Díaz-Sánchez, Angel G; Macías-Rubalcava, Martha L; Hermoso, Juan A; Rodríguez-Romero, Adela

    2014-10-01

    Plants express chitinase and chitinase-like proteins (CLPs) belonging to the glycosyl hydrolases of the GH18 and GH19 families, which exhibit varied functions. CLPs in the GH18 family have been structurally and functionally characterized; however, there are no structures available for any member of the GH19 family. In this study, two CLPs of the GH19 family from the rubber tree Hevea brasiliensis (HbCLP1 and HbCLP2) were cloned, expressed and characterized. HbCLP1 was identical to the allergen Hev b 11.0101 previously described by others, while HbCLP2 was a novel isoform exhibiting an unusual half chitin-binding domain before the catalytic domain. Sequence alignments showed that in the two proteins the catalytic residues Glu117 and Glu147 in HbCLP1 and HbCLP2, respectively, were mutated to Ala, accounting for the lack of activity. Nonetheless, both CLPs bound chitin and chitotriose (GlcNAc)3 with high affinities, as evaluated with chitin-affinity chromatography and tryptophan fluorescence experiments. The chitin-binding domains also bound chitotriose with even higher affinities. The crystal structures of the HbCLP1-isolated domains were determined at high resolution. The analysis of the crystallographic models and docking experiments using (GlcNAc)6 oligosaccharides provides evidence of the residues involved in sugar binding. Endochitinase activity was restored in both proteins by mutating residues A117E (HbCLP1) and A147E (HbCLP2); the distance between the catalytic proton donor and the catalytic nucleophile in the in silico mutated residues was 9.5 Å, as occurs in inverting enzymes. HbCLP1 and HbCLP2 were highly thermostable and exhibited antifungal activity against Alternaria alternata, suggesting their participation in plant defense mechanisms. PMID:25104038

  11. Identification of Powdery Mildew Responsive Genes in Hevea brasiliensis through mRNA Differential Display

    PubMed Central

    Li, Xiang; Bi, Zhenghong; Di, Rong; Liang, Peng; He, Qiguang; Liu, Wenbo; Miao, Weiguo; Zheng, Fucong

    2016-01-01

    Powdery mildew is an important disease of rubber trees caused by Oidium heveae B. A. Steinmann. As far as we know, none of the resistance genes related to powdery mildew have been isolated from the rubber tree. There is little information available at the molecular level regarding how a rubber tree develops defense mechanisms against this pathogen. We have studied rubber tree mRNA transcripts from the resistant RRIC52 cultivar by differential display analysis. Leaves inoculated with the spores of O. heveae were collected from 0 to 120 hpi in order to identify pathogen-regulated genes at different infection stages. We identified 78 rubber tree genes that were differentially expressed during the plant–pathogen interaction. BLAST analysis for these 78 ESTs classified them into seven functional groups: cell wall and membrane pathways, transcription factor and regulatory proteins, transporters, signal transduction, phytoalexin biosynthesis, other metabolism functions, and unknown functions. The gene expression for eight of these genes was validated by qRT-PCR in both RRIC52 and the partially susceptible Reyan 7-33-97 cultivars, revealing the similar or differential changes of gene expressions between these two cultivars. This study has improved our overall understanding of the molecular mechanisms of rubber tree resistance to powdery mildew. PMID:26840302

  12. Identification of Powdery Mildew Responsive Genes in Hevea brasiliensis through mRNA Differential Display.

    PubMed

    Li, Xiang; Bi, Zhenghong; Di, Rong; Liang, Peng; He, Qiguang; Liu, Wenbo; Miao, Weiguo; Zheng, Fucong

    2016-01-01

    Powdery mildew is an important disease of rubber trees caused by Oidium heveae B. A. Steinmann. As far as we know, none of the resistance genes related to powdery mildew have been isolated from the rubber tree. There is little information available at the molecular level regarding how a rubber tree develops defense mechanisms against this pathogen. We have studied rubber tree mRNA transcripts from the resistant RRIC52 cultivar by differential display analysis. Leaves inoculated with the spores of O. heveae were collected from 0 to 120 hpi in order to identify pathogen-regulated genes at different infection stages. We identified 78 rubber tree genes that were differentially expressed during the plant-pathogen interaction. BLAST analysis for these 78 ESTs classified them into seven functional groups: cell wall and membrane pathways, transcription factor and regulatory proteins, transporters, signal transduction, phytoalexin biosynthesis, other metabolism functions, and unknown functions. The gene expression for eight of these genes was validated by qRT-PCR in both RRIC52 and the partially susceptible Reyan 7-33-97 cultivars, revealing the similar or differential changes of gene expressions between these two cultivars. This study has improved our overall understanding of the molecular mechanisms of rubber tree resistance to powdery mildew. PMID:26840302

  13. Micro-organisms in latex and natural rubber coagula of Hevea brasiliensis and their impact on rubber composition, structure and properties.

    PubMed

    Salomez, M; Subileau, M; Intapun, J; Bonfils, F; Sainte-Beuve, J; Vaysse, L; Dubreucq, E

    2014-10-01

    Natural rubber, produced by coagulation of the latex from the tree Hevea brasiliensis, is an important biopolymer used in many applications for its outstanding properties. Besides polyisoprene, latex is rich in many nonisoprene components such as carbohydrates, proteins and lipids and thereby constitutes a favourable medium for the development of micro-organisms. The fresh rubber coagula obtained by latex coagulation are not immediately processed, allowing the development of various microbial communities. The time period between tree tapping and coagula processing is called maturation, during which an evolution of the properties of the corresponding dry natural rubber occurs. This evolution is partly related to the activity of micro-organisms and to the modification of the biochemical composition. This review synthesizes the current knowledge on microbial populations in latex and natural rubber coagula of H. brasiliensis and the changes they induce on the biochemistry and technical properties of natural rubber during maturation. PMID:24891014

  14. Carbon isotope composition of latex does not reflect temporal variations of photosynthetic carbon isotope discrimination in rubber trees (Hevea brasiliensis).

    PubMed

    Kanpanon, Nicha; Kasemsap, Poonpipope; Thaler, Philippe; Kositsup, Boonthida; Gay, Frédéric; Lacote, Régis; Epron, Daniel

    2015-11-01

    Latex, the cytoplasm of laticiferous cells localized in the inner bark of rubber trees (Hevea brasiliensis Müll. Arg.), is collected by tapping the bark. Following tapping, latex flows out of the trunk and is regenerated, whereas in untapped trees, there is no natural exudation. It is still unknown whether the carbohydrates used for latex regeneration in tapped trees is coming from recent photosynthates or from stored carbohydrates, and in the former case, it is expected that latex carbon isotope composition of tapped trees will vary seasonally, whereas latex isotope composition of untapped trees will be more stable. Temporal variations of carbon isotope composition of trunk latex (δ(13)C-L), leaf soluble compounds (δ(13)C-S) and bulk leaf material (δ(13)C-B) collected from tapped and untapped 20-year-old trees were compared. A marked difference in δ(13)C-L was observed between tapped and untapped trees whatever the season. Trunk latex from tapped trees was more depleted (1.6‰ on average) with more variable δ(13)C values than those of untapped trees. δ(13)C-L was higher and more stable across seasons than δ(13)C-S and δ(13)C-B, with a maximum seasonal difference of 0.7‰ for tapped trees and 0.3‰ for untapped trees. δ(13)C-B was lower in tapped than in untapped trees, increasing from August (middle of the rainy season) to April (end of the dry season). Differences in δ(13)C-L and δ(13)C-B between tapped and untapped trees indicated that tapping affects the metabolism of both laticiferous cells and leaves. The lack of correlation between δ(13)C-L and δ(13)C-S suggests that recent photosynthates are mixed in the large pool of stored carbohydrates that are involved in latex regeneration after tapping. PMID:26358051

  15. Identification, Functional Study, and Promoter Analysis of HbMFT1, a Homolog of MFT from Rubber Tree (Hevea brasiliensis).

    PubMed

    Bi, Zhenghong; Li, Xiang; Huang, Huasun; Hua, Yuwei

    2016-01-01

    A homolog of MOTHER OF FT AND TFL1 (MFT) was isolated from Hevea brasiliensis and its biological function was investigated. Protein multiple sequence alignment and phylogenetic analysis revealed that HbMFT1 conserved critical amino acid residues to distinguish MFT, FLOWERING LOCUS T (FT) and TERMINAL FLOWER1 (TFL1)-like proteins and showed a closer genetic relationship to the MFT-like group. The accumulation of HbMFT1 was generally detected in various tissues except pericarps, with the highest expression in embryos and relatively higher expression in roots and stems of seedlings, flowering inflorescences, and male and female flowers. HbMFT1 putative promoter analysis showed that tissue-specific, environmental change responsive and hormone-signaling responsive elements were generally present. HbMFT1 was strongly induced under a short-day condition at 28 °C, with the highest expression after the onset of a day. Overexpression of HbMFT1 inhibited seed germination, seedling growth, and flowering in transgenic Arabidopsis. The qRT-PCR further confirmed that APETALA1 (AP1) and FRUITFULL (FUL) were drastically down-regulated in 35S::HbMFT1 plants. A histochemical β-glucuronidase (GUS) assay showed that HbMFT1::GUS activity was mainly detected in stamens and mature seeds coinciding with its original expression and notably induced in rosette leaves and seedlings of transgenic Arabidopsis by exogenous abscisic acid (ABA) due to the presence of ABA cis-elements in HbMFT1 promoter. These results suggested that HbMFT1 was mainly involved in maintenance of seed maturation and stamen development, but negatively controlled germination, growth and development of seedlings and flowering. In addition, the HbMFT1 promoter can be utilized in controlling transgene expression in stamens and seeds of rubber tree or other plant species. PMID:26950112

  16. The Hevea brasiliensis XIP aquaporin subfamily: genomic, structural and functional characterizations with relevance to intensive latex harvesting.

    PubMed

    Lopez, David; Amira, Maroua Ben; Brown, Daniel; Muries, Beatriz; Brunel-Michac, Nicole; Bourgerie, Sylvain; Porcheron, Benoit; Lemoine, Remi; Chrestin, Hervé; Mollison, Ewan; Di Cola, Alessandra; Frigerio, Lorenzo; Julien, Jean-Louis; Gousset-Dupont, Aurélie; Fumanal, Boris; Label, Philippe; Pujade-Renaud, Valérie; Auguin, Daniel; Venisse, Jean-Stéphane

    2016-07-01

    X-Intrinsic Proteins (XIP) were recently identified in a narrow range of plants as a full clade within the aquaporins. These channels reportedly facilitate the transport of a wide range of hydrophobic solutes. The functional roles of XIP in planta remain poorly identified. In this study, we found three XIP genes (HbXIP1;1, HbXIP2;1 and HbXIP3;1) in the Hevea brasiliensis genome. Comprehensive bioinformatics, biochemical and structural analyses were used to acquire a better understanding of this AQP subfamily. Phylogenetic analysis revealed that HbXIPs clustered into two major groups, each distributed in a specific lineage of the order Malpighiales. Tissue-specific expression profiles showed that only HbXIP2;1 was expressed in all the vegetative tissues tested (leaves, stem, bark, xylem and latex), suggesting that HbXIP2;1 could take part in a wide range of cellular processes. This is particularly relevant to the rubber-producing laticiferous system, where this isoform was found to be up-regulated during tapping and ethylene treatments. Furthermore, the XIP transcriptional pattern is significantly correlated to latex production level. Structural comparison with SoPIP2;1 from Spinacia oleracea species provides new insights into the possible role of structural checkpoints by which HbXIP2;1 ensures glycerol transfer across the membrane. From these results, we discuss the physiological involvement of glycerol and HbXIP2;1 in water homeostasis and carbon stream of challenged laticifers. The characterization of HbXIP2;1 during rubber tree tapping lends new insights into molecular and physiological response processes of laticifer metabolism in the context of latex exploitation. PMID:27068521

  17. Identification, Functional Study, and Promoter Analysis of HbMFT1, a Homolog of MFT from Rubber Tree (Hevea brasiliensis)

    PubMed Central

    Bi, Zhenghong; Li, Xiang; Huang, Huasun; Hua, Yuwei

    2016-01-01

    A homolog of MOTHER OF FT AND TFL1 (MFT) was isolated from Hevea brasiliensis and its biological function was investigated. Protein multiple sequence alignment and phylogenetic analysis revealed that HbMFT1 conserved critical amino acid residues to distinguish MFT, FLOWERING LOCUS T (FT) and TERMINAL FLOWER1 (TFL1)-like proteins and showed a closer genetic relationship to the MFT-like group. The accumulation of HbMFT1 was generally detected in various tissues except pericarps, with the highest expression in embryos and relatively higher expression in roots and stems of seedlings, flowering inflorescences, and male and female flowers. HbMFT1 putative promoter analysis showed that tissue-specific, environmental change responsive and hormone-signaling responsive elements were generally present. HbMFT1 was strongly induced under a short-day condition at 28 °C, with the highest expression after the onset of a day. Overexpression of HbMFT1 inhibited seed germination, seedling growth, and flowering in transgenic Arabidopsis. The qRT-PCR further confirmed that APETALA1 (AP1) and FRUITFULL (FUL) were drastically down-regulated in 35S::HbMFT1 plants. A histochemical β-glucuronidase (GUS) assay showed that HbMFT1::GUS activity was mainly detected in stamens and mature seeds coinciding with its original expression and notably induced in rosette leaves and seedlings of transgenic Arabidopsis by exogenous abscisic acid (ABA) due to the presence of ABA cis-elements in HbMFT1 promoter. These results suggested that HbMFT1 was mainly involved in maintenance of seed maturation and stamen development, but negatively controlled germination, growth and development of seedlings and flowering. In addition, the HbMFT1 promoter can be utilized in controlling transgene expression in stamens and seeds of rubber tree or other plant species. PMID:26950112

  18. Quantitative expression analysis of drought responsive genes in clones of Hevea with varying levels of drought tolerance.

    PubMed

    Luke, Lisha P; Mohamed Sathik, M B; Thomas, Molly; Kuruvilla, Linu; Sumesh, K V; Annamalainathan, K

    2015-04-01

    In order to meet the ever rising global demand for natural rubber, cultivation of Hevea is being extended to non-traditional regions of India where extreme climatic conditions like drought and low temperature negatively influence the crop performance. In order to ensure maximum productivity, identification of drought tolerant clones of Hevea which can cope up with stress and give better crop yield is essential. Several attempts have been made previously to identify genes that are associated with drought tolerance in Hevea. In the present study, quantitative expression analysis was made using quantitative PCR for seven drought associated transcripts in four clones of Hevea with varying levels of drought tolerance. Among the seven genes studied, Mitogen Activated Protein (MAP) kinase, Myeloblastosis (Myb) transcription factor, C-repeat responsive element/Dehydration Responsive Element (CRT/DRE) binding factor and Nuclear Factor Y subunit A (NFYA) showed a positive association with drought tolerance. Transcripts of ascorbate peroxidase and heat shock protein 70 (HSP 70) did not show any correlation with drought tolerance. Interestingly, catalase gene was found down regulated in all the clones under drought condition. The possible role of these genes based on their level of gene expression in four different clones of Hevea with varying levels of drought tolerance is discussed. PMID:25964712

  19. Whole-transcriptome survey of the putative ATP-binding cassette (ABC) transporter family genes in the latex-producing laticifers of Hevea brasiliensis.

    PubMed

    Zhiyi, Nie; Guijuan, Kang; Yu, Li; Longjun, Dai; Rizhong, Zeng

    2015-01-01

    The ATP-binding cassette (ABC) proteins or transporters constitute a large protein family in plants and are involved in many different cellular functions and processes, including solute transportation, channel regulation and molecular switches, etc. Through transcriptome sequencing, a transcriptome-wide survey and expression analysis of the ABC protein genes were carried out using the laticiferous latex from Hevea brasiliensis (rubber tree). A total of 46 putative ABC family proteins were identified in the H. brasiliensis latex. These consisted of 12 'full-size', 21 'half-size' and 13 other putative ABC proteins, and all of them showed strong conservation with their Arabidopsis thaliana counterparts. This study indicated that all eight plant ABC protein paralog subfamilies were identified in the H. brasiliensis latex, of which ABCB, ABCG and ABCI were the most abundant. Real-time quantitative reverse transcription-polymerase chain reaction assays demonstrated that gene expression of several latex ABC proteins was regulated by ethylene, jasmonic acid or bark tapping (a wound stress) stimulation, and that HbABCB15, HbABCB19, HbABCD1 and HbABCG21 responded most significantly of all to the abiotic stresses. The identification and expression analysis of the latex ABC family proteins could facilitate further investigation into their physiological involvement in latex metabolism and rubber biosynthesis by H. brasiliensis. PMID:25615936

  20. Whole-Transcriptome Survey of the Putative ATP-Binding Cassette (ABC) Transporter Family Genes in the Latex-Producing Laticifers of Hevea brasiliensis

    PubMed Central

    Zhiyi, Nie; Guijuan, Kang; Yu, Li; Longjun, Dai; Rizhong, Zeng

    2015-01-01

    The ATP-binding cassette (ABC) proteins or transporters constitute a large protein family in plants and are involved in many different cellular functions and processes, including solute transportation, channel regulation and molecular switches, etc. Through transcriptome sequencing, a transcriptome-wide survey and expression analysis of the ABC protein genes were carried out using the laticiferous latex from Hevea brasiliensis (rubber tree). A total of 46 putative ABC family proteins were identified in the H. brasiliensis latex. These consisted of 12 ‘full-size’, 21 ‘half-size’ and 13 other putative ABC proteins, and all of them showed strong conservation with their Arabidopsis thaliana counterparts. This study indicated that all eight plant ABC protein paralog subfamilies were identified in the H. brasiliensis latex, of which ABCB, ABCG and ABCI were the most abundant. Real-time quantitative reverse transcription-polymerase chain reaction assays demonstrated that gene expression of several latex ABC proteins was regulated by ethylene, jasmonic acid or bark tapping (a wound stress) stimulation, and that HbABCB15, HbABCB19, HbABCD1 and HbABCG21 responded most significantly of all to the abiotic stresses. The identification and expression analysis of the latex ABC family proteins could facilitate further investigation into their physiological involvement in latex metabolism and rubber biosynthesis by H. brasiliensis. PMID:25615936

  1. Seasonal and clonal variations in technological and thermal properties of raw Hevea natural rubber

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study was undertaken over a ten-month period, under the environmental conditions within the state of Mato Grosso, Brazil, to evaluate the causes of variation in technological and thermal properties of raw natural rubber from different clones of Hevea brasiliensis (GT 1, PR 255, FX 3864 and RRIM...

  2. Evapotranspiration of rubber (Hevea brasiliensis) under the highly seasonal rainfall regime of the Asian monsoon in mainland Southeast Asia

    NASA Astrophysics Data System (ADS)

    Giambelluca, T. W.; Mudd, R. G.; Liu, W.; Kobayashi, N.; Ziegler, A. D.; Miyazawa, Y.; Kumagai, T.; Huang, M.

    2012-12-01

    The Asian Monsoon dominates the climate of the mainland Southeast Asia (MSEA) region, characterized by a highly seasonal rainfall regime in which 80-90% of annual rainfall occurs during the 6-month (May-October) wet season. The accompanying extremes in soil moisture, solar radiation, and vapor pressure deficit exert strong controls on ecosystem fluxes, including evapotranspiration (ET). Rubber (Hevea brasiliensis), the major commercial crop currently replacing traditional agriculture and secondary forests in MSEA is a native of the equatorial Amazon rainforest, and differs physiologically from the dominant native SE Asian forest tree species. It sheds its leaves in the middle of the dry season and flushes new leaves before the onset of the wet season. In some areas, rubber cultivation is suspected of having caused changes in local climate and watershed processes, including a dramatic downward trend in fog frequency and large increases in surface runoff and soil erosion (Wu et al., 2001, Int. J. Sust. Dev. World Ecol. 8:337-345). Guardiola-Claramonte et al. (2008, Ecohydrology 1:13-22; 2010, Ecohydrology 3:306-314) noted striking differences in the timing and rate of dry season root-water extraction under rubber as compared with other vegetation types. To investigate the environmental impacts of rubber, eddy covariance flux towers were installed to monitor energy, water, and carbon exchange at rubber plantation sites in northeastern Thailand and Cambodia. Results of the first two years of observations at the sites indicate that controls on ET differ between wet and dry seasons, with varying responses to energy, soil moisture, canopy wetness, and leaf area. Despite the long dry season and loss of leaves for several weeks, rubber accumulates exceptionally high annual ET totals, exceeding those of natural forest and other plant functional types in the region. The phenology of rubber represents a disruption of the land-atmosphere interactions of native and other non

  3. Identification of differentially expressed genes and signalling pathways in bark of Hevea brasiliensis seedlings associated with secondary laticifer differentiation using gene expression microarray.

    PubMed

    Loh, Swee Cheng; Thottathil, Gincy P; Othman, Ahmad Sofiman

    2016-10-01

    The natural rubber of Para rubber tree, Hevea brasiliensis, is the main crop involved in industrial rubber production due to its superior quality. The Hevea bark is commercially exploited to obtain latex, which is produced from the articulated secondary laticifer. The laticifer is well defined in the aspect of morphology; however, only some genes associated with its development have been reported. We successfully induced secondary laticifer in the jasmonic acid (JA)-treated and linolenic acid (LA)-treated Hevea bark but secondary laticifer is not observed in the ethephon (ET)-treated and untreated Hevea bark. In this study, we analysed 27,195 gene models using NimbleGen microarrays based on the Hevea draft genome. 491 filtered differentially expressed (FDE) transcripts that are common to both JA- and LA-treated bark samples but not ET-treated bark samples were identified. In the Eukaryotic Orthologous Group (KOG) analysis, 491 FDE transcripts belong to different functional categories that reflect the diverse processes and pathways involved in laticifer differentiation. In the Kyoto Encyclopedia of Genes and Genomes (KEGG) and KOG analysis, the profile of the FDE transcripts suggest that JA- and LA-treated bark samples have a sufficient molecular basis for secondary laticifer differentiation, especially regarding secondary metabolites metabolism. FDE genes in this category are from the cytochrome (CYP) P450 family, ATP-binding cassette (ABC) transporter family, short-chain dehydrogenase/reductase (SDR) family, or cinnamyl alcohol dehydrogenase (CAD) family. The data includes many genes involved in cell division, cell wall synthesis, and cell differentiation. The most abundant transcript in FDE list was SDR65C, reflecting its importance in laticifer differentiation. Using the Basic Local Alignment Search Tool (BLAST) as part of annotation and functional prediction, several characterised as well as uncharacterized transcription factors and genes were found in the

  4. Increasing the reaction rate of hydroxynitrile lyase from Hevea brasiliensis toward mandelonitrile by copying active site residues from an esterase that accepts aromatic esters.

    PubMed

    von Langermann, Jan; Nedrud, David M; Kazlauskas, Romas J

    2014-09-01

    The natural substrate of hydroxynitrile lyase from rubber tree (HbHNL, Hevea brasiliensis) is acetone cyanohydrin, but synthetic applications usually involve aromatic cyanohydrins such as mandelonitrile. To increase the activity of HbHNL toward this unnatural substrate, we replaced active site residues in HbHNL with the corresponding ones from esterase SABP2 (salicylic acid binding protein 2). Although this enzyme catalyzes a different reaction (hydrolysis of esters), its natural substrate (methyl salicylate) contains an aromatic ring. Three of the eleven single-amino-acid-substitution variants of HbHNL reacted more rapidly with mandelonitrile. The best was HbHNL-L121Y, with a kcat 4.2 times higher and high enantioselectivity. Site-saturation mutagenesis at position 121 identified three other improved variants. We hypothesize that the smaller active site orients the aromatic substrate more productively. PMID:25044660

  5. Expression Profiles, Characterization and Function of HbTCTP in Rubber Tree (Hevea brasiliensis)

    PubMed Central

    Deng, Zhi; Chen, Jiangshu; Leclercq, Julie; Zhou, Zhuangzhi; Liu, Changren; Liu, Hui; Yang, Hong; Montoro, Pascal; Xia, Zhihui; Li, Dejun

    2016-01-01

    As a highly conserved protein, the translationally controlled tumor protein (TCTP) carries out vital roles in various life processes. In rubber tree, two TCTP genes, HbTCTP and HbTCTP1, were cloned, but only HbTCTP1 was studied in details. In this study, cis-acting regulatory elements, expression patterns, subcellular localization, interacting proteins, and antioxidant activity of HbTCTP were systematically analyzed. Besides the common cis-acting regulatory elements, HbTCTP promoter also harbored various known cis-elements that respond to hormone/stresses. Being consistent with the aforementioned results, HbTCTP was regulated by drought, low temperature, high salt, ethylene (ET), wounding, H2O2, and methyl jasmonate (MeJA) treatments. HbTCTP was expressed throughout different tissues and developmental stages of leaves. In addition, HbTCTP was associated with tapping panel dryness (TPD). HbTCTP was localized in the membrane, cytoplasm and the nucleus, and interacted with four proteins rubber elongation factor (REF), 17.5 kDa heat shock family protein, annexin, and REF-like stress related protein 1. Being similar to HbTCTP1, HbTCTP also indicated antioxidant activity in metal-catalyzed oxidation (MCO) system. Our results are useful for further understanding the molecular characterization and expression profiles of HbTCTP, but also lay a solid foundation for elucidating the function of HbTCTP in rubber tree. PMID:27375647

  6. Comparative Analysis of Latex Transcriptome Reveals Putative Molecular Mechanisms Underlying Super Productivity of Hevea brasiliensis

    PubMed Central

    Li, Heping; Fan, Yujie; Yang, Jianghua; Qi, Jiyan; Li, Huibo

    2013-01-01

    Increasing demand for natural rubber prompts studies into the mechanisms governing the productivity of rubber tree (Heveabrasiliensis). It is very interesting to notice that a rubber tree of clone PR107 in Yunnan, China is reported to yield more than 20 times higher than the average rubber tree. This super-high-yielding (SHY) rubber tree (designated as SY107), produced 4.12 kg of latex (cytoplasm of rubber producing laticifers, containing about 30% of rubber) per tapping, more than 7-fold higher than that of the control. This rubber tree is therefore a good material to study how the rubber production is regulated at a molecular aspect. A comprehensive cDNA-AFLP transcript profiling was performed on the latex of SY107 and its average counterparts by using the 384 selective primer pairs for two restriction enzyme combinations (ApoI/MseI and TaqI/MseI). A total of 746 differentially expressed (DE) transcript-derived fragments (TDFs) were identified, of which the expression patterns of 453 TDFs were further confirmed by RT-PCR. These RT-PCR confirmed TDFs represented 352 non-redundant genes, of which 215 had known or partially known functions and were grouped into 10 functional categories. The top three largest categories were transcription and protein synthesis (representing 24.7% of the total genes), defense and stress (15.3%), and primary and secondary metabolism (14.0%). Detailed analysis of the DE-genes suggests notable characteristics of SHY phenotype in improved sucrose loading capability, rubber biosynthesis-preferred sugar utilization, enhanced general metabolism and timely stress alleviation. However, the SHY phenotype has little correlation with rubber-biosynthesis pathway genes. PMID:24066172

  7. Applicability of Landsat TM data for inventorying and monitoring of rubber (Hevea brasiliensis) plantations in Selangor, Malaysia: Linkages to policies

    NASA Astrophysics Data System (ADS)

    Suratman, Mohd Nazip

    2003-06-01

    Rubber tree (Hevea brasiliensis (Wild ex Adr. De Juss) Muell Arg.) plantations in Malaysia are important sources of natural rubber and wood products. Effective management and appropriate policy for these resources require reliable information on resource dynamics and forecasts of resource availability. The need for inventories and monitoring systems prompted this research into utilising ground information and satellite imagery for developing methods for forest plantation inventory. Monitoring procedures were developed using three dates of Landsat Thematic Mapper (TM) imagery. The specific objectives of the research were: (1) to develop an effective method for inventorying rubber tree plantations using an appropriate combination of satellite imagery and ground sampling in the State of Selangor, Malaysia; (2) to demonstrate the application of a Landsat TM-based rubber volume model in an extended area of rubber plantations south of Kuala Lumpur (KL), Malaysia; (3) to develop an operational methodology for monitoring land use/cover change, with a primary focus on rubber plantations; and (4) to identify relationships between the primary drivers of resource change and policies, and examine the evidence of policies---rubber area change linkages in the study area. Reasonably accurate predictions of the volume, age, and area of rubber plantations were obtained from Landsat TM data. The use of supervised image classification and an image segmentation approach for rubber volume model application showed better performance for volume prediction than a combined land use/cover and rubber volume classification technique, thus providing a useful tool for displaying rubber stand volume within segments or spatial units across the landscape. The combined use of a time series of Landsat TM imagery, modified postclassification change detection, and geographic information system (GIS) techniques made it possible to produce land use/cover change matrices and rubber area change information

  8. The cyanogenic syndrome in rubber tree Hevea brasiliensis: tissue-damage-dependent activation of linamarase and hydroxynitrile lyase accelerates hydrogen cyanide release

    PubMed Central

    Kadow, Daniel; Voß, Karsten; Selmar, Dirk; Lieberei, Reinhard

    2012-01-01

    Background and Aims The release of hydrogen cyanide (HCN) from injured plant tissue affects multiple ecological interactions. Plant-derived HCN can act as a defence against herbivores and also plays an important role in plant–pathogen interactions. Crucial for activity as a feeding deterrent is the amount of HCN generated per unit time, referred to as cyanogenic capacity (HCNc). Strong intraspecific variation in HCNc has been observed among cyanogenic plants. This variation, in addition to genotypic variability (e.g. in Trifolium repens), can result from modifications in the expression level of the enzymes involved in either cyanogenic precursor formation or HCN release (as seen in Sorghum bicolor and Phaseolus lunatus). Thus, a modification or modulation of HCNc in reaction to the environment can only be achieved from one to the next generation when under genetic control and within days or hours when transcriptional regulations are involved. In the present study, it is shown that in rubber tree (Hevea brasiliensis) HCNc is modulated by post-translational activity regulation of the key enzymes for cyanide release. Methods Linamarase (LIN) and hydroxynitrile lyase (HNL) activity was determined by colorimetric assays utilizing dissociation of the substrates p-nitrophenyl-β-d-glucopyranoside and acetone cyanohydrin, respectively. Key Results In rubber tree leaves, LIN and HNL show up to ten-fold increased activity in response to tissue damage. This enzyme activation occurs within seconds and results in accelerated HCN formation. It is restricted to the damaged leaf area and depends on the severity of tissue damage. Conclusions LIN and HNL activation (in contrast to genetic and transcriptional regulations) allows an immediate, local and damage type-dependent modulation of the cyanogenic response. Accordingly, this post-translational activation plays a decisive role in the defence of H. brasiliensis against herbivores as well as pathogens and may allow more flexible

  9. Structural analysis of the endogenous glycoallergen Hev b 2 (endo-β-1,3-glucanase) from Hevea brasiliensis and its recognition by human basophils

    PubMed Central

    Rodríguez-Romero, Adela; Hernández-Santoyo, Alejandra; Fuentes-Silva, Deyanira; Palomares, Laura A.; Muñoz-Cruz, Samira; Yépez-Mulia, Lilian; Orozco-Martínez, Socorro

    2014-01-01

    Endogenous glycosylated Hev b 2 (endo-β-1,3-glucanase) from Hevea brasiliensis is an important latex allergen that is recognized by IgE antibodies from patients who suffer from latex allergy. The carbohydrate moieties of Hev b 2 constitute a potentially important IgE-binding epitope that could be responsible for its cross-reactivity. Here, the structure of the endogenous isoform II of Hev b 2 that exhibits three post-translational modifications, including an N-terminal pyro­glutamate and two glycosylation sites at Asn27 and at Asn314, is reported from two crystal polymorphs. These modifications form a patch on the surface of the molecule that is proposed to be one of the binding sites for IgE. A structure is also proposed for the most important N-glycan present in this protein as determined by digestion with specific enzymes. To analyze the role of the carbohydrate moieties in IgE antibody binding and in human basophil activation, the glycoallergen was enzymatically deglycosylated and evaluated. Time-lapse automated video microscopy of basophils stimulated with glycosylated Hev b 2 revealed basophil activation and degranulation. Immunological studies suggested that carbohydrates on Hev b 2 represent an allergenic IgE epitope. In addition, a dimer was found in each asymmetric unit that may reflect a regulatory mechanism of this plant defence protein. PMID:24531467

  10. Functional Characterization of Hevea brasiliensis CRT/DRE Binding Factor 1 Gene Revealed Regulation Potential in the CBF Pathway of Tropical Perennial Tree.

    PubMed

    Cheng, Han; Cai, Haibin; Fu, Haitian; An, Zewei; Fang, Jialin; Hu, Yanshi; Guo, Dianjing; Huang, Huasun

    2015-01-01

    Rubber trees (Hevea brasiliensis) are susceptible to low temperature and therefore are only planted in the tropical regions. In the past few decades, although rubber trees have been successfully planted in the northern margin of tropical area in China, they suffered from cold injury during the winter. To understand the physiological response under cold stress, we isolated a C-repeat binding factor 1 (CBF1) gene from the rubber tree. This gene (HbCBF1) was found to respond to cold stress but not drought or ABA stress. The corresponding HbCBF1 protein showed CRT/DRE binding activity in gel shift experiment. To further characterize its molecular function, the HbCBF1 gene was overexpressed in Arabidopsis. The HbCBF1 over expression (OE) line showed enhanced cold resistance and relatively slow dehydration, and the expression of Arabidopsis CBF pathway downstream target genes, e.g. AtCOR15a and AtRD29a, were significantly activated under non-acclimation condition. These data suggest HbCBF1 gene is a functional member of the CBF gene family, and may play important regulation function in rubber tree. PMID:26361044

  11. A full picture of enzymatic catalysis by hydroxynitrile lyases from Hevea brasiliensis: protonation dependent reaction steps and residue-gated movement of the substrate and the product.

    PubMed

    Zhao, Yuan; Chen, Nanhao; Mo, Yirong; Cao, Zexing

    2014-12-28

    Hydroxynitrile lyases (HNLs) defend plants from herbivores and microbial attack by releasing cyanide from hydroxynitriles. The reverse process has been productively applied to bioorganic syntheses of pharmaceuticals and agrochemicals. To improve our understanding of the catalytic mechanism of HNLs, extensive ab initio QM/MM and classical MM molecular dynamics simulations have been performed to explore the catalytic conversion of cyanohydrins into aldehyde (or ketone) and HCN by hydroxynitrile lyases from Hevea brasiliensis (HbHNLs). It was found that the catalytic reaction approximately follows a two-stage mechanism. The first stage involves two fast processes including the proton abstraction of the substrate through a double-proton transfer and the C-CN bond cleavage, while the second stage concerns HCN formation and is rate-determining. The complete free energy profile exhibits a peak of ∼18 kcal mol(-1). Interestingly, the protonation state of Lys236 influences the efficiency of the enzyme only to some extent, but it changes the entire catalytic mechanism. The dynamical behaviors of substrate delivery and HCN release are basically modulated by the gate movement of Trp128. The remarkable exothermicity of substrate binding and the facile release of HCN may drive the enzyme-catalyzed reaction to proceed along the substrate decomposition efficiently. Computational mutagenesis reveals the key residues which play an important role in the catalytic process. PMID:25375265

  12. Isolation of a furan fatty acid from Hevea brasiliensis latex employing the combined use of pH-zone-refining and conventional countercurrent chromatography.

    PubMed

    Englert, Michael; Ulms, Kerstin; Wendlinger, Christine; Vetter, Walter

    2016-02-01

    Furan fatty acids are valuable and bioactive minor fatty acids that usually contribute <0.1% to the fatty acid content of food samples. Their biological role still remains unclear as authentic furan fatty acid standards are not readily available and thorough experimental studies verifying the relevance of furan fatty acids are thus virtually impossible. An efficient protocol for the isolation of the furan fatty acid 9-(3-methyl-5-pentylfuran-2-yl)-nonanoic acid from hydrolyzed and centrifuged latex of Hevea brasiliensis was developed using countercurrent chromatography. A first run using pH-zone-refining countercurrent chromatography provided 48.4 mg of 9-(3-methyl-5-pentylfuran-2-yl)-nonanoic acid from 210 mg latex extract in a purity of 95%. The purity was increased to 99% by means of one second run in conventional countercurrent chromatography mode. The Structure and purity of 9-(3-methyl-5-pentylfuran-2-yl)-nonanoic acid were determined by gas chromatography coupled to mass spectrometry and (1) H and (13) C NMR spectroscopy. PMID:26611161

  13. Simulation of Canopy CO2/H2O Fluxes for a Rubber (Hevea Brasiliensis) Plantation in Central Cambodia: The Effect of the Regular Spacing of Planted Trees

    SciTech Connect

    Kumagai, Tomo'omi; Mudd, Ryan; Miyazawa, Yoshiyuki; Liu, Wen; Giambelluca, Thomas; Kobayashi, N.; Lim, Tiva Khan; Jomura, Mayuko; Matsumoto, Kazuho; Huang, Maoyi; Chen, Qi; Ziegler, Alan; Yin, Song

    2013-09-10

    We developed a soil-vegetation-atmosphere transfer (SVAT) model applicable to simulating CO2 and H2O fluxes from the canopies of rubber plantations, which are characterized by distinct canopy clumping produced by regular spacing of plantation trees. Rubber (Hevea brasiliensis Müll. Arg.) plantations, which are rapidly expanding into both climatically optimal and sub-optimal environments throughout mainland Southeast Asia, potentially change the partitioning of water, energy, and carbon at multiple scales, compared with traditional land covers it is replacing. Describing the biosphere-atmosphere exchange in rubber plantations via SVAT modeling is therefore essential to understanding the impacts on environmental processes. The regular spacing of plantation trees creates a peculiar canopy structure that is not well represented in most SVAT models, which generally assumes a non-uniform spacing of vegetation. Herein we develop a SVAT model applicable to rubber plantation and an evaluation method for its canopy structure, and examine how the peculiar canopy structure of rubber plantations affects canopy CO2 and H2O exchanges. Model results are compared with measurements collected at a field site in central Cambodia. Our findings suggest that it is crucial to account for intensive canopy clumping in order to reproduce observed rubber plantation fluxes. These results suggest a potentially optimal spacing of rubber trees to produce high productivity and water use efficiency.

  14. Functional Characterization of Hevea brasiliensis CRT/DRE Binding Factor 1 Gene Revealed Regulation Potential in the CBF Pathway of Tropical Perennial Tree

    PubMed Central

    Cheng, Han; Cai, Haibin; Fu, Haitian; An, Zewei; Fang, Jialin; Hu, Yanshi; Guo, Dianjing; Huang, Huasun

    2015-01-01

    Rubber trees (Hevea brasiliensis) are susceptible to low temperature and therefore are only planted in the tropical regions. In the past few decades, although rubber trees have been successfully planted in the northern margin of tropical area in China, they suffered from cold injury during the winter. To understand the physiological response under cold stress, we isolated a C-repeat binding factor 1 (CBF1) gene from the rubber tree. This gene (HbCBF1) was found to respond to cold stress but not drought or ABA stress. The corresponding HbCBF1 protein showed CRT/DRE binding activity in gel shift experiment. To further characterize its molecular function, the HbCBF1 gene was overexpressed in Arabidopsis. The HbCBF1 over expression (OE) line showed enhanced cold resistance and relatively slow dehydration, and the expression of Arabidopsis CBF pathway downstream target genes, e.g. AtCOR15a and AtRD29a, were significantly activated under non-acclimation condition. These data suggest HbCBF1 gene is a functional member of the CBF gene family, and may play important regulation function in rubber tree. PMID:26361044

  15. [Mites (Acari) from rubber trees (Hevea brasiliensis Muell. Arg., Euphorbiaceae) and spontaneous euphorbiaceous in rubber trees cultivation].

    PubMed

    Bellini, Marcos R; Feres, Reinaldo J F; Buosi, Renato

    2008-01-01

    Quarterly samples were done in 2001 on three rubber tree plantation in the northwest of the state of São Paulo. Three rubber trees of each locality were sampled. Between the rows of rubber tree four species of spontaneous euphorbiaceous were collected: Chamaesyce hirta, C. hyssopifolia, Euphorbia heterophylla and Phyllanthus tenellus. A total of 8.954 mites of 38 species, belonging to 31 genera of 11 families were collected. Tydeidae and Phytoseiidae had the highest diversity of species, 9 and 7, respectively. The most abundant families were Eriophyidae (3.594), Tydeidae (2.825) and Tenuipalpidae (1.027). The most abundant species on the rubber trees were: phytophagous - Calacarus heveae Feres, Tenuipalpus heveae Baker, Lorryia sp.2, Lorryia formosa Cooreman and Lorryia sp.1; predators - Zetzellia quasagistemas Hernandes & Feres, Pronematus sp., Iphiseiodes zuluagai Denmark & Muma and Euseius citrifolius Denmark & Muma. Among the spontaneous euphorbiaceous, predatory mites were abundantly found on C. hirta and E. heterophylla, mainly Pronematus sp. and E. citrifolius, suggesting that these plants could be important in the maintenance of these predators in the rubber tree cultivation areas. However, plants that can shelter predators and at the same time exert strong competition (nutrients, water etc) to rubber trees, can not be recommended for pest management programs. Studies about competition between rubber trees and spontaneous plants need to be conducted for feasible efficient programs of environmental management, aiming at the control of pest mites of rubber tree. PMID:18813750

  16. Construction of a high-density integrated genetic linkage map of rubber tree (Hevea brasiliensis) using genotyping-by-sequencing (GBS)

    PubMed Central

    Pootakham, Wirulda; Ruang-Areerate, Panthita; Jomchai, Nukoon; Sonthirod, Chutima; Sangsrakru, Duangjai; Yoocha, Thippawan; Theerawattanasuk, Kanikar; Nirapathpongporn, Kanlaya; Romruensukharom, Phayao; Tragoonrung, Somvong; Tangphatsornruang, Sithichoke

    2015-01-01

    Construction of linkage maps is crucial for genetic studies and marker-assisted breeding programs. Recent advances in next generation sequencing technologies allow for the generation of high-density linkage maps, especially in non-model species lacking extensive genomic resources. Here, we constructed a high-density integrated genetic linkage map of rubber tree (Hevea brasiliensis), the sole commercial producer of high-quality natural rubber. We applied a genotyping-by-sequencing (GBS) technique to simultaneously discover and genotype single nucleotide polymorphism (SNP) markers in two rubber tree populations. A total of 21,353 single nucleotide substitutions were identified, 55% of which represented transition events. GBS-based genetic maps of populations P and C comprised 1704 and 1719 markers and encompassed 2041 cM and 1874 cM, respectively. The average marker densities of these two maps were one SNP in 1.23–1.25 cM. A total of 1114 shared SNP markers were used to merge the two component maps. An integrated linkage map consisted of 2321 markers and spanned the cumulative length of 2052 cM. The composite map showed a substantial improvement in marker density, with one SNP marker in every 0.89 cM. To our knowledge, this is the most saturated genetic map in rubber tree to date. This integrated map allowed us to anchor 28,965 contigs, covering 135 Mb or 12% of the published rubber tree genome. We demonstrated that GBS is a robust and cost-effective approach for generating a common set of genome-wide SNP data suitable for constructing integrated linkage maps from multiple populations in a highly heterozygous agricultural species. PMID:26074933

  17. Construction of a high-density integrated genetic linkage map of rubber tree (Hevea brasiliensis) using genotyping-by-sequencing (GBS).

    PubMed

    Pootakham, Wirulda; Ruang-Areerate, Panthita; Jomchai, Nukoon; Sonthirod, Chutima; Sangsrakru, Duangjai; Yoocha, Thippawan; Theerawattanasuk, Kanikar; Nirapathpongporn, Kanlaya; Romruensukharom, Phayao; Tragoonrung, Somvong; Tangphatsornruang, Sithichoke

    2015-01-01

    Construction of linkage maps is crucial for genetic studies and marker-assisted breeding programs. Recent advances in next generation sequencing technologies allow for the generation of high-density linkage maps, especially in non-model species lacking extensive genomic resources. Here, we constructed a high-density integrated genetic linkage map of rubber tree (Hevea brasiliensis), the sole commercial producer of high-quality natural rubber. We applied a genotyping-by-sequencing (GBS) technique to simultaneously discover and genotype single nucleotide polymorphism (SNP) markers in two rubber tree populations. A total of 21,353 single nucleotide substitutions were identified, 55% of which represented transition events. GBS-based genetic maps of populations P and C comprised 1704 and 1719 markers and encompassed 2041 cM and 1874 cM, respectively. The average marker densities of these two maps were one SNP in 1.23-1.25 cM. A total of 1114 shared SNP markers were used to merge the two component maps. An integrated linkage map consisted of 2321 markers and spanned the cumulative length of 2052 cM. The composite map showed a substantial improvement in marker density, with one SNP marker in every 0.89 cM. To our knowledge, this is the most saturated genetic map in rubber tree to date. This integrated map allowed us to anchor 28,965 contigs, covering 135 Mb or 12% of the published rubber tree genome. We demonstrated that GBS is a robust and cost-effective approach for generating a common set of genome-wide SNP data suitable for constructing integrated linkage maps from multiple populations in a highly heterozygous agricultural species. PMID:26074933

  18. Genome-Wide Identification of Jatropha curcas Aquaporin Genes and the Comparative Analysis Provides Insights into the Gene Family Expansion and Evolution in Hevea brasiliensis.

    PubMed

    Zou, Zhi; Yang, Lifu; Gong, Jun; Mo, Yeyong; Wang, Jikun; Cao, Jianhua; An, Feng; Xie, Guishui

    2016-01-01

    Aquaporins (AQPs) are channel-forming integral membrane proteins that transport water and other small solutes across biological membranes. Despite the vital role of AQPs, to date, little is known in physic nut (Jatropha curcas L., Euphorbiaceae), an important non-edible oilseed crop with great potential for the production of biodiesel. In this study, 32 AQP genes were identified from the physic nut genome and the family number is relatively small in comparison to 51 in another Euphorbiaceae plant, rubber tree (Hevea brasiliensis Muell. Arg.). Based on the phylogenetic analysis, the JcAQPs were assigned to five subfamilies, i.e., nine plasma membrane intrinsic proteins (PIPs), nine tonoplast intrinsic proteins (TIPs), eight NOD26-like intrinsic proteins (NIPs), two X intrinsic proteins (XIPs), and four small basic intrinsic proteins (SIPs). Like rubber tree and other plant species, functional prediction based on the aromatic/arginine selectivity filter, Froger's positions, and specificity-determining positions showed a remarkable difference in substrate specificity among subfamilies of JcAQPs. Genome-wide comparative analysis revealed the specific expansion of PIP and TIP subfamilies in rubber tree and the specific gene loss of the XIP subfamily in physic nut. Furthermore, by analyzing deep transcriptome sequencing data, the expression evolution especially the expression divergence of duplicated HbAQP genes was also investigated and discussed. Results obtained from this study not only provide valuable information for future functional analysis and utilization of Jc/HbAQP genes, but also provide a useful reference to survey the gene family expansion and evolution in Euphorbiaceae plants and other plant species. PMID:27066041

  19. Influence of low-level laser associated with osteogenic proteins recombinant human BMP-2 and Hevea brasiliensis on bone repair in Wistar rats.

    PubMed

    Iyomasa, Mamie Mizusaki; Issa, João Paulo Mardegan; de Queiróz Tavares, Mirella Lemos; Pereira, Yamba Carla Lara; Stuani, Maria Bernadete Sasso; Mishima, Fabíola; Coutinho-Netto, Joaquim; Sebald, Walter

    2012-02-01

    This study analyzed the newly formed bone tissue after application of recombinant human BMP-2 (rhBMP-2) and P-1 (extracted from Hevea brasiliensis) proteins, 2 weeks after the creation of a critical bone defect in male Wistar rats treated or not with a low-intensity laser (GaAlAs 780 nm, 60 mW of power, and energy density dose of 30 J/cm(2)). The animals were divided into two major groups: (1) bone defect plus low-intensity laser treatment and (2) bone defect without laser irradiation. The following subgroups were also analyzed: (a) 5 μg of pure rhBMP-2; (b) 5 μg of pure P-1 fraction; (c) 5 μg of rhBMP-2/monoolein gel; (d) 5 μg of P-1 fraction/monoolein gel; (e) pure monoolein gel. Comparisons of the groups receiving laser treatment with those that did not receive laser irradiation show differences in the areas of new bone tissue. The group treated with 5 μg of rhBMP-2 and laser irradiation was not significantly different (P >0.05) than the nonirradiated group that received the same treatment. The irradiated, rhBMP-2/monoolein gel treatment group showed a lower area of bone formation than the nonirradiated, rhBMP-2/gel monoolein treatment group (P < 0.001). The area of new bone tissue in the other nonirradiated and irradiated groups was not significantly different (P > 0.05). Furthermore, the group that received the 5 μg of rhBMP-2 application showed the greatest bone formation. We conclude that the laser treatment did not interfere with the area of new bone tissue growth and that the greatest stimulus for bone formation involved application of the rhBMP-2 protein. PMID:21761492

  20. Genome-Wide Identification of Jatropha curcas Aquaporin Genes and the Comparative Analysis Provides Insights into the Gene Family Expansion and Evolution in Hevea brasiliensis

    PubMed Central

    Zou, Zhi; Yang, Lifu; Gong, Jun; Mo, Yeyong; Wang, Jikun; Cao, Jianhua; An, Feng; Xie, Guishui

    2016-01-01

    Aquaporins (AQPs) are channel-forming integral membrane proteins that transport water and other small solutes across biological membranes. Despite the vital role of AQPs, to date, little is known in physic nut (Jatropha curcas L., Euphorbiaceae), an important non-edible oilseed crop with great potential for the production of biodiesel. In this study, 32 AQP genes were identified from the physic nut genome and the family number is relatively small in comparison to 51 in another Euphorbiaceae plant, rubber tree (Hevea brasiliensis Muell. Arg.). Based on the phylogenetic analysis, the JcAQPs were assigned to five subfamilies, i.e., nine plasma membrane intrinsic proteins (PIPs), nine tonoplast intrinsic proteins (TIPs), eight NOD26-like intrinsic proteins (NIPs), two X intrinsic proteins (XIPs), and four small basic intrinsic proteins (SIPs). Like rubber tree and other plant species, functional prediction based on the aromatic/arginine selectivity filter, Froger's positions, and specificity-determining positions showed a remarkable difference in substrate specificity among subfamilies of JcAQPs. Genome-wide comparative analysis revealed the specific expansion of PIP and TIP subfamilies in rubber tree and the specific gene loss of the XIP subfamily in physic nut. Furthermore, by analyzing deep transcriptome sequencing data, the expression evolution especially the expression divergence of duplicated HbAQP genes was also investigated and discussed. Results obtained from this study not only provide valuable information for future functional analysis and utilization of Jc/HbAQP genes, but also provide a useful reference to survey the gene family expansion and evolution in Euphorbiaceae plants and other plant species. PMID:27066041

  1. Structural analysis of the endogenous glycoallergen Hev b 2 (endo-β-1,3-glucanase) from Hevea brasiliensis and its recognition by human basophils

    SciTech Connect

    Rodríguez-Romero, Adela Hernández-Santoyo, Alejandra; Fuentes-Silva, Deyanira; Palomares, Laura A.; Muñoz-Cruz, Samira; Yépez-Mulia, Lilian; Orozco-Martínez, Socorro

    2014-02-01

    This study describes the three-dimensional structure of the endogenous glycosylated allergen Hev b 2 (endo-β-1,3-glucanase), which exhibits three post-translational modifications that form a patch on the surface of the molecule that is proposed to be an allergenic IgE epitope. Endogenous glycosylated Hev b 2 (endo-β-1,3-glucanase) from Hevea brasiliensis is an important latex allergen that is recognized by IgE antibodies from patients who suffer from latex allergy. The carbohydrate moieties of Hev b 2 constitute a potentially important IgE-binding epitope that could be responsible for its cross-reactivity. Here, the structure of the endogenous isoform II of Hev b 2 that exhibits three post-translational modifications, including an N-terminal pyroglutamate and two glycosylation sites at Asn27 and at Asn314, is reported from two crystal polymorphs. These modifications form a patch on the surface of the molecule that is proposed to be one of the binding sites for IgE. A structure is also proposed for the most important N-glycan present in this protein as determined by digestion with specific enzymes. To analyze the role of the carbohydrate moieties in IgE antibody binding and in human basophil activation, the glycoallergen was enzymatically deglycosylated and evaluated. Time-lapse automated video microscopy of basophils stimulated with glycosylated Hev b 2 revealed basophil activation and degranulation. Immunological studies suggested that carbohydrates on Hev b 2 represent an allergenic IgE epitope. In addition, a dimer was found in each asymmetric unit that may reflect a regulatory mechanism of this plant defence protein.

  2. Uncovering divergent evolution of α/β-hydrolases: a surprising residue substitution needed to convert Hevea brasiliensis hydroxynitrile lyase into an esterase.

    PubMed

    Nedrud, David M; Lin, Hui; Lopez, Gilsinia; Padhi, Santosh K; Legatt, Graig A; Kaz-Lauskas, Romas J

    2014-11-01

    Hevea brasiliensis hydroxynitrile lyase (HbHNL) and salicylic acid binding protein 2 (SABP2, an esterase) share 45% amino acid sequence identity, the same protein fold, and even the same catalytic triad of Ser-His-Asp. However, they catalyze different reactions: cleavage of hydroxynitriles and hydrolysis of esters, respectively. To understand how other active site differences in the two enzymes enable the same catalytic triad to catalyze different reactions, we substituted amino acid residues in HbHNL with the corresponding residues from SABP2, expecting hydroxynitrile lyase activity to decrease and esterase activity to increase. Previous mechanistic studies and x-ray crystallography suggested that esterase activity requires removal of an active site lysine and threonine from the hydroxynitrile lyase. The Thr11Gly Lys236Gly substitutions in HbHNL reduced hydroxynitrile lyase activity for cleavage of mandelonitrile 100-fold, but increased esterase activity only threefold to kcat ~ 0.1 min(-1) for hydrolysis of p-nitrophenyl acetate. Adding a third substitution - Glu79His - increased esterase activity more than tenfold to kcat ~ 1.6 min(-1). The specificity constant (kcat/KM) for this triple substitution variant versus wild type HbHNL shifted more than one million-fold from hydroxynitrile lyase activity (acetone cyanohydrin substrate) to esterase activity (p-nitrophenyl acetate substrate). The contribution of Glu79His to esterase activity was surprising since esterases and lipases contain many different amino acids at this position, including glutamate. Saturation mutagenesis at position 79 showed that 13 of 19 possible amino acid substitutions increased esterase activity, suggesting that removal of glutamate, not addition of histidine, increased esterase activity. Molecular modeling indicates that Glu79 disrupts esterase activity in HbHNL when its negatively charged side chain distorts the orientation of the catalytic histidine. Naturally occurring glutamate at the

  3. Population Dynamics of the Rubber Plantation Litter Beetle Luprops tristis, in Relation to Annual Cycle of Foliage Phenology of Its Host, the Para Rubber Tree, Hevea brasiliensis

    PubMed Central

    Sabu, Thomas K.; Vinod, K.V.

    2009-01-01

    The population dynamics of the rubber plantation litter beetle, Luprops tristis Fabricius 1801 (Coleoptera: Tenebrionidae) was assessed in relation to the phenology of leaf shedding and defoliation pattern of para rubber trees, Hevea brasiliensis Müll. Arg (Malpighiales: Euphorbiaceae), during a two year study period. The abundance of adults, larvae and pupae per 1m2 of litter sample was recorded. Post dormancy beetles appeared in leaf litter following annual leaf shedding, whereas larvae, pupae and teneral adults were present after leaf flush. No stages were recorded from plantations following the summer rains until the annual litter fall in the next season. Parental adults peaked at the time of leaf sprouting and tender leaf fall. Larvae and teneral adults peaked at the time of premature fall of green leaves and flowers. Teneral adults of six age classes were recorded and all entered dormancy irrespective of the feeding time available to each age class. Females outnumbered males in the parent generation, while the sex ratio of new generation adults was not biased towards either sex. The phenological stages of rubber trees included leaf fall in late December and early January, leaf sprouting and new leaf production in January and flowering in February. All feeding stages of L. tristis peaked in abundance when premature leaves are most abundant in the leaf litter. Prediction of the timing of appearance of various developmental stages of L. tristis in plantations, invasion into buildings and intensity of population build up in rubber belts is possible by tracking the phenology of leaf fall in rubber plantations, time of return of post dormancy adults and the onset of summer rainfall. Perfect synchrony was recorded between the field return of parental adults with annual leaf shedding, the oviposition phase of parental adults with tender leaf fall at the time of leaf sprouting, and larval and teneral adult stages with premature fall of leaves. Premature leaf

  4. Leaf ecophysiology of rubber (Hevea brasiliensis) and its implication to stand-level transpiration in the Asian monsoon in mainland Southeast Asia

    NASA Astrophysics Data System (ADS)

    Miyazawa, Y.; Kobayashi, N.; Mudd, R. G.; Yin, S.; LimKhan, T.; Giambelluca, T. W.; Kumagai, T.

    2012-12-01

    In Indochina Peninsula, rubber (Hevea brasiliensis) plantation is expanding rapidly as the major commercial crop. Rubber, which is a native of Amazon rainforests, are thought to have transpiration characteristics different from other native species in this region, which is characterized by high rainfall occurs in rainy season and severe drought in dry season. Studies suggested high stand transpiration rate (E) in rubber plantations, possibly due to the water use even in dry seasons or to high leaf photosynthetic capacity. Despite recent advances about the leaf-level measurements in leaf gas exchange traits, knowledge has been collected without clear linkage to the seasonal trends in stand-level gas exchange and its seasonal trends. In this study, we monitored the leaf-level photosynthetic capacity (Vcmax25) and a parameter for stomatal control (m, Ball et al. 1987) to show whether changes in atmospheric environments and leaf gas exchange traits explain the high E and its seasonal trends (annual peak in mid rainy season). Results showed that rubber had Vcmax25 and m similar to temperate broadleaved tree species and higher than other species in Indochina Peninsula, indicating that high Vcmax25 would be responsible for the higher E than other forests in Indochina Peninsula. Vcmax25 reached an annual maximum soon after leaf expansion before the beginning of rainy season, keep the level until soon before leaf fall in mid dry season and did not supported the hypothesis that these parameters increased in mid rainy season. The m also showed slight seasonal variations without clear trends. Multilayer model well modeled the diurnal trends in measured E using sap flow measurements, except soon after leaf expansion. Constant leaf gas exchange rates and good reproduction of finer time scale E suggested that increased E in mid rainy season was not due to the increased evaporative demand or changes in leaf gas exchange rates, but to the increased LAI. Poor modeling soon after

  5. How do rubber (Hevea brasiliensis) plantations behave under seasonal water stress in northeastern Thailand and central Cambodia?

    SciTech Connect

    Kumagai, Tomo'omi; Mudd, Ryan G.; Giambelluca, Thomas W.; Kobayashi, Nakako; Miyazawa, Yoshiyuki; Lim, Tiva Khan; Liu, Wen; Huang, Maoyi; Fox, Jefferson M.; Ziegler, Alan D.; Yin, Song; Mak, Sophea Veasna; Kasemsap, Poonpipope

    2015-11-01

    Plantation rubber (Hevea brasiliensis Müll. Arg.) is a viable economic resource for Southeast Asian countries. Consequently, rubber plantations are rapidly expanding into both climatically optimal and sub-optimal environments throughout mainland Southeast Asia, potentially changing the partitioning of water, energy, and carbon at multiple scales, compared with the traditional land covers they are replacing. Delineating the characteristics of biosphere-atmosphere exchange in rubber plantations is therefore important to understanding the impacts of such land use change on environmental processes. We have conducted eddy flux measurements in two rubber plantation sites: (1) Som Sanuk (SS), located northern Thailand; and (2) Cambodian Rubber Research Institute (CRRI), central Cambodia. Both sites have a distinct dry season. Measurements were made over a 3-year period. We used combination of actual evapotranspiration (ET) flux measurements and an inversed version of a simple 2-layer ET model for estimating the mean canopy stomatal conductances (gs), which is among the most effective measures for describing water and energy exchanges and tree water use characteristics. A main novelty in this analysis is that the rubber canopy conductance can be extracted from total surface conductance (including the canopy and the vegetation floor effects) and hence environmental and biological controls on rubber tree gs are explicitly compared at each site in different seasons and years. It is demonstrated how each studied rubber plantation copes with each strong seasonal drought via tree water use strategies. Potential tree water use deficit (precipitation (P) – potential evaporation (ET_POT)) for each season (i.e., December-February: DJF, March-May: MAM, June-August: JJA, and September-November: SON) revealed in which season and how the water use should be controlled. We found that in seasons when actual tree water use deficit (P

  6. Regulation of HbPIP2;3, a Latex-Abundant Water Transporter, Is Associated with Latex Dilution and Yield in the Rubber Tree (Hevea brasiliensis Muell. Arg.).

    PubMed

    An, Feng; Zou, Zhi; Cai, Xiuqing; Wang, Jin; Rookes, James; Lin, Weifu; Cahill, David; Kong, Lingxue

    2015-01-01

    Rubber tree (Hevea brasiliensis) latex, the source of natural rubber, is synthesised in the cytoplasm of laticifers. Efficient water inflow into laticifers is crucial for latex flow and production since it is the determinant of the total solid content of latex and its fluidity after tapping. As the mature laticifer vessel rings are devoid of plasmodesmata, water exchange between laticifers and surrounding cells is believed to be governed by plasma membrane intrinsic proteins (PIPs). To identify the most important PIP aquaporin in the water balance of laticifers, the transcriptional profiles of ten-latex-expressed PIPs were analysed. One of the most abundant transcripts, designated HbPIP2;3, was characterised in this study. When tested in Xenopus laevis oocytes HbPIP2;3 showed a high efficiency in increasing plasmalemma water conductance. Expression analysis indicated that the HbPIP2;3 gene was preferentially expressed in latex, and the transcripts were up-regulated by both wounding and exogenously applied Ethrel (a commonly-used ethylene releaser). Although regular tapping up-regulated the expression of HbPIP2;3 during the first few tappings of the virginal rubber trees, the transcriptional kinetics of HbPIP2;3 to Ethrel stimulation in the regularly tapped tree exhibited a similar pattern to that of the previously reported HbPIP2;1 in the virginal rubber trees. Furthermore, the mRNA level of HbPIP2;3 was associated with clonal yield potential and the Ethrel stimulation response. Together, these results have revealed the central regulatory role of HbPIP2;3 in laticifer water balance and ethylene stimulation of latex production in Hevea. PMID:25927524

  7. Regulation of HbPIP2;3, a Latex-Abundant Water Transporter, Is Associated with Latex Dilution and Yield in the Rubber Tree (Hevea brasiliensis Muell. Arg.)

    PubMed Central

    Cai, Xiuqing; Wang, Jin; Rookes, James; Lin, Weifu; Cahill, David; Kong, Lingxue

    2015-01-01

    Rubber tree (Hevea brasiliensis) latex, the source of natural rubber, is synthesised in the cytoplasm of laticifers. Efficient water inflow into laticifers is crucial for latex flow and production since it is the determinant of the total solid content of latex and its fluidity after tapping. As the mature laticifer vessel rings are devoid of plasmodesmata, water exchange between laticifers and surrounding cells is believed to be governed by plasma membrane intrinsic proteins (PIPs). To identify the most important PIP aquaporin in the water balance of laticifers, the transcriptional profiles of ten-latex-expressed PIPs were analysed. One of the most abundant transcripts, designated HbPIP2;3, was characterised in this study. When tested in Xenopus laevis oocytes HbPIP2;3 showed a high efficiency in increasing plasmalemma water conductance. Expression analysis indicated that the HbPIP2;3 gene was preferentially expressed in latex, and the transcripts were up-regulated by both wounding and exogenously applied Ethrel (a commonly-used ethylene releaser). Although regular tapping up-regulated the expression of HbPIP2;3 during the first few tappings of the virginal rubber trees, the transcriptional kinetics of HbPIP2;3 to Ethrel stimulation in the regularly tapped tree exhibited a similar pattern to that of the previously reported HbPIP2;1 in the virginal rubber trees. Furthermore, the mRNA level of HbPIP2;3 was associated with clonal yield potential and the Ethrel stimulation response. Together, these results have revealed the central regulatory role of HbPIP2;3 in laticifer water balance and ethylene stimulation of latex production in Hevea. PMID:25927524

  8. Mites (Arachnida: Acari) collected on rubber trees Hevea brasiliensis (Willd. ex A.Juss.) Müll.Arg. in Santana, Amapá state, Brazil.

    PubMed

    Deus, E G; Souza, M S M; Mineiro, J L C; Adaime, R; Santos, R S

    2012-11-01

    The purpose of this study was to elaborate a preliminary list of the mite species associated with rubber trees in the municipality of Santana, in the state of Amapá, Brazil. Two collections of rubber tree leaves were conducted on May 2nd and June 5th , 2010. Twenty-five plants were sampled at random. Three leaves were collected per plant, from the lower third of the crown. The samples were placed in paper bags, packed in an isothermal box chilled gel-based pulp plant (Gelo-X(®)), and transported to the Entomology Laboratory at Embrapa Amapá, in Macapá. The leaflets were examined under a stereomicroscope, and the mites found on the adaxial and abaxial surfaces of the leaves were collected with a stilet, mounted on microscope slides in Hoyer's medium, and later identified. We collected a total of 1,722 mites of 10 families: Acaridae, Cunaxidae, Eriophyidae, Iolinidae, Phytoseiidae, Stigmaeidae, Tarsonemidae, Tenuipalpidae, Tydeidae, and Winterschmidtiidae, in addition to unidentified species of the suborders Oribatida and Astigmatina. The family Phytoseiidae represented only 2.90% of specimens collected, but showed the highest species richness (5 species). The only representative of Tenuipalpidae was Tenuipalpus heveae Baker, 1945, but 81.13% of the mites collected in this study belonged to this species. PMID:23295522

  9. Hevea Linamarase—A Nonspecific β-Glycosidase 1

    PubMed Central

    Selmar, Dirk; Lieberei, Reinhard; Biehl, Böle; Voigt, Jürgen

    1987-01-01

    In the leaf tissue of the cyanogenic plant Hevea brasiliensis, which contains large amounts of linamarin, there is no specific linamarase. In Hevea leaves only one β-glucosidase is detectable. It is responsible for the cleavage of all β-glucosides and β-galactosides occurring in Hevea leaf tissue, including the cyanogenic glucoside linamarin. Therefore, the enzyme is referred to as a β-glycosidase instead of the term β-glucosidase. This β-glycosidase has a broad substrate spectrum and occurs in multiple forms. These homo-oligomeric forms are interconvertible by dissociation-association processes. The monomer is a single protein of 64 kilodaltons. PMID:16665288

  10. Hevea Linamarase-A Nonspecific beta-Glycosidase.

    PubMed

    Selmar, D; Lieberei, R; Biehl, B; Voigt, J

    1987-03-01

    In the leaf tissue of the cyanogenic plant Hevea brasiliensis, which contains large amounts of linamarin, there is no specific linamarase. In Hevea leaves only one beta-glucosidase is detectable. It is responsible for the cleavage of all beta-glucosides and beta-galactosides occurring in Hevea leaf tissue, including the cyanogenic glucoside linamarin. Therefore, the enzyme is referred to as a beta-glycosidase instead of the term beta-glucosidase. This beta-glycosidase has a broad substrate spectrum and occurs in multiple forms. These homo-oligomeric forms are interconvertible by dissociation-association processes. The monomer is a single protein of 64 kilodaltons. PMID:16665288

  11. QTL Mapping of Growth-Related Traits in a Full-Sib Family of Rubber Tree (Hevea brasiliensis) Evaluated in a Sub-Tropical Climate

    PubMed Central

    Mantello, Camila Campos; Silva, Carla Cristina; Garcia, Dominique; Le Guen, Vincent; Cardoso, Saulo Emilio Almeida; Garcia, Antonio Augusto Franco; Souza, Anete Pereira

    2013-01-01

    The rubber tree (Hevea spp.), cultivated in equatorial and tropical countries, is the primary plant used in natural rubber production. Due to genetic and physiological constraints, inbred lines of this species are not available. Therefore, alternative approaches are required for the characterization of this species, such as the genetic mapping of full-sib crosses derived from outbred parents. In the present study, an integrated genetic map was obtained for a full-sib cross family with simple sequence repeats (SSRs) and expressed sequence tag (EST-SSR) markers, which can display different segregation patterns. To study the genetic architecture of the traits related to growth in two different conditions (winter and summer), quantitative trait loci (QTL) mapping was also performed using the integrated map. Traits evaluated were height and girth growth, and the statistical model was based in an extension of composite interval mapping. The obtained molecular genetic map has 284 markers distributed among 23 linkage groups with a total length of 2688.8 cM. A total of 18 QTLs for growth traits during the summer and winter seasons were detected. A comparison between the different seasons was also conducted. For height, QTLs detected during the summer season were different from the ones detected during winter season. This type of difference was also observed for girth. Integrated maps are important for genetics studies in outbred species because they represent more accurately the polymorphisms observed in the genitors. QTL mapping revealed several interesting findings, such as a dominance effect and unique segregation patterns that each QTL could exhibit, which were independent of the flanking markers. The QTLs identified in this study, especially those related to phenotypic variation associated with winter could help studies of marker-assisted selection that are particularly important when the objective of a breeding program is to obtain phenotypes that are adapted to sub

  12. Seasonal Patterns of Fine Root Production and Turnover in a Mature Rubber Tree (Hevea brasiliensis Müll. Arg.) Stand- Differentiation with Soil Depth and Implications for Soil Carbon Stocks

    PubMed Central

    Maeght, Jean-Luc; Gonkhamdee, Santimaitree; Clément, Corentin; Isarangkool Na Ayutthaya, Supat; Stokes, Alexia; Pierret, Alain

    2015-01-01

    Fine root dynamics is a main driver of soil carbon stocks, particularly in tropical forests, yet major uncertainties still surround estimates of fine root production and turnover. This lack of knowledge is largely due to the fact that studying root dynamics in situ, particularly deep in the soil, remains highly challenging. We explored the interactions between fine root dynamics, soil depth, and rainfall in mature rubber trees (Hevea brasiliensis Müll. Arg.) exposed to sub-optimal edaphic and climatic conditions. A root observation access well was installed in northern Thailand to monitor root dynamics along a 4.5 m deep soil profile. Image-based measurements of root elongation and lifespan of individual roots were carried out at monthly intervals over 3 years. Soil depth was found to have a significant effect on root turnover. Surprisingly, root turnover increased with soil depth and root half-life was 16, 6–8, and only 4 months at 0.5, 1.0, 2.5, and 3.0 m deep, respectively (with the exception of roots at 4.5 m which had a half-life similar to that found between depths of 1.0 and 2.5 m). Within the first two meters of the soil profile, the highest rates of root emergence occurred about 3 months after the onset of the rainy season, while deeper in the soil, root emergence was not linked to the rainfall pattern. Root emergence was limited during leaf flushing (between March and May), particularly within the first two meters of the profile. Between soil depths of 0.5 and 2.0 m, root mortality appeared independent of variations in root emergence, but below 2.0 m, peaks in root emergence and death were synchronized. Shallow parts of the root system were more responsive to rainfall than their deeper counterparts. Increased root emergence in deep soil toward the onset of the dry season could correspond to a drought acclimation mechanism, with the relative importance of deep water capture increasing once rainfall ceased. The considerable soil depth regularly explored

  13. Seasonal Patterns of Fine Root Production and Turnover in a Mature Rubber Tree (Hevea brasiliensis Müll. Arg.) Stand- Differentiation with Soil Depth and Implications for Soil Carbon Stocks.

    PubMed

    Maeght, Jean-Luc; Gonkhamdee, Santimaitree; Clément, Corentin; Isarangkool Na Ayutthaya, Supat; Stokes, Alexia; Pierret, Alain

    2015-01-01

    Fine root dynamics is a main driver of soil carbon stocks, particularly in tropical forests, yet major uncertainties still surround estimates of fine root production and turnover. This lack of knowledge is largely due to the fact that studying root dynamics in situ, particularly deep in the soil, remains highly challenging. We explored the interactions between fine root dynamics, soil depth, and rainfall in mature rubber trees (Hevea brasiliensis Müll. Arg.) exposed to sub-optimal edaphic and climatic conditions. A root observation access well was installed in northern Thailand to monitor root dynamics along a 4.5 m deep soil profile. Image-based measurements of root elongation and lifespan of individual roots were carried out at monthly intervals over 3 years. Soil depth was found to have a significant effect on root turnover. Surprisingly, root turnover increased with soil depth and root half-life was 16, 6-8, and only 4 months at 0.5, 1.0, 2.5, and 3.0 m deep, respectively (with the exception of roots at 4.5 m which had a half-life similar to that found between depths of 1.0 and 2.5 m). Within the first two meters of the soil profile, the highest rates of root emergence occurred about 3 months after the onset of the rainy season, while deeper in the soil, root emergence was not linked to the rainfall pattern. Root emergence was limited during leaf flushing (between March and May), particularly within the first two meters of the profile. Between soil depths of 0.5 and 2.0 m, root mortality appeared independent of variations in root emergence, but below 2.0 m, peaks in root emergence and death were synchronized. Shallow parts of the root system were more responsive to rainfall than their deeper counterparts. Increased root emergence in deep soil toward the onset of the dry season could correspond to a drought acclimation mechanism, with the relative importance of deep water capture increasing once rainfall ceased. The considerable soil depth regularly explored by

  14. Paracoccidioides brasiliensis presents two different cDNAs encoding homologues of the fructose 1,6-biphosphate aldolase: protein isolation, cloning of the cDNAs and genes, structural, phylogenetic, and expression analysis.

    PubMed

    Carneiro, Lílian Carla; de Faria, Fabrícia P; Felipe, M Sueli S; Pereira, Maristela; de Almeida Soares, Célia M

    2005-01-01

    A proteomic approach was used to identify a 39 kDa antigen of Paracoccidioides brasiliensis. Amino acid sequences of the N-terminal and of endoproteinase Lys-C digested peptides revealed the protein to be a fructose 1,6-biphosphate aldolase (FBA) Class II of P. brasiliensis. Two cDNA homologues, Pbfba1 and Pbfba2, were cloned and characterized. Pbfba1 encoded a predicted polypeptide of 360 amino acids that was highly homologous in the primary structure to the same enzyme from fungi and bacteria. The other DNA, Pbfba2, encoded a polypeptide predicted to be 363 amino acids. The sequence of Pbfba2 differed significantly from Pbfba1. Phylogenetic and molecular analysis supports the concept of gene duplication for FBAs in P. brasiliensis, constituting a two-member family. Expression analysis demonstrated differential expression for both fbas genes in P. brasiliensis cells. PMID:15588996

  15. Evaluation of Reference Genes for Quantitative Real-Time PCR Analysis of the Gene Expression in Laticifers on the Basis of Latex Flow in Rubber Tree (Hevea brasiliensis Muell. Arg.)

    PubMed Central

    Chao, Jinquan; Yang, Shuguang; Chen, Yueyi; Tian, Wei-Min

    2016-01-01

    Latex exploitation-caused latex flow is effective in enhancing latex regeneration in laticifer cells of rubber tree. It should be suitable for screening appropriate reference gene for analysis of the expression of latex regeneration-related genes by quantitative real-time PCR (qRT-PCR). In the present study, the expression stability of 23 candidate reference genes was evaluated on the basis of latex flow by using geNorm and NormFinder algorithms. Ubiquitin-protein ligase 2a (UBC2a) and ubiquitin-protein ligase 2b (UBC2b) were the two most stable genes among the selected candidate references in rubber tree clones with differential duration of latex flow. The two genes were also high-ranked in previous reference gene screening across different tissues and experimental conditions. By contrast, the transcripts of latex regeneration-related genes fluctuated significantly during latex flow. The results suggest that screening reference gene during latex flow should be an efficient and effective clue for selection of reference genes in qRT-PCR. PMID:27524995

  16. Evaluation of Reference Genes for Quantitative Real-Time PCR Analysis of the Gene Expression in Laticifers on the Basis of Latex Flow in Rubber Tree (Hevea brasiliensis Muell. Arg.).

    PubMed

    Chao, Jinquan; Yang, Shuguang; Chen, Yueyi; Tian, Wei-Min

    2016-01-01

    Latex exploitation-caused latex flow is effective in enhancing latex regeneration in laticifer cells of rubber tree. It should be suitable for screening appropriate reference gene for analysis of the expression of latex regeneration-related genes by quantitative real-time PCR (qRT-PCR). In the present study, the expression stability of 23 candidate reference genes was evaluated on the basis of latex flow by using geNorm and NormFinder algorithms. Ubiquitin-protein ligase 2a (UBC2a) and ubiquitin-protein ligase 2b (UBC2b) were the two most stable genes among the selected candidate references in rubber tree clones with differential duration of latex flow. The two genes were also high-ranked in previous reference gene screening across different tissues and experimental conditions. By contrast, the transcripts of latex regeneration-related genes fluctuated significantly during latex flow. The results suggest that screening reference gene during latex flow should be an efficient and effective clue for selection of reference genes in qRT-PCR. PMID:27524995

  17. Cloning

    MedlinePlus

    Cloning describes the processes used to create an exact genetic replica of another cell, tissue or organism. ... named Dolly. There are three different types of cloning: Gene cloning, which creates copies of genes or ...

  18. Cloning

    MedlinePlus

    ... DNA Reproductive cloning, which creates copies of whole animals Therapeutic cloning, which creates embryonic stem cells. Researchers hope to use these cells to grow healthy tissue to replace injured or diseased tissues in the human body. NIH: National Human Genome Research Institute

  19. Cloning and functional characterization of β-1, 3-glucanase gene from Podophyllum hexandrum - a high altitude Himalayan plant.

    PubMed

    Dogra, Vivek; Sreenivasulu, Yelam

    2015-01-01

    Podophyllum hexandrum is a high-altitude medicinal plant exploited for its etoposides which are potential anticancer compounds. ß-1, 3-glucanase cDNA was cloned from the germinating seeds of Podophyllum (Ph-glucanase). Glucanases belong to pathogenesis related glycohydralase family of proteins, which also play an important role in endosperm weakening and testa rupture during seed germination. Analysis of cloned nucleotide sequence revealed Ph-glucanase with an open reading frame of 852bp encoding a protein of 283 amino acids with a molecular mass of 31kDa and pI of 4.39. In-silico structure prediction of Ph-glucanase showed homology with that of Hevea brasiliensis (3em5B). Structural stability and enhanced catalytic efficiency in harsh climatic conditions possibly due to the presence of glycosyl hydrolase motif (LGIVISESGWPSAG) and a connecting loop towards inner side and well exposed carbohydrate metabolism domain-COG5309, can readily hydrolyse cell wall sugar moieties. Seeds from the transgenic Arabidopsis plants over-expressing Ph-glucanase showed better germination performance against a wide range of temperatures and abscisic acid (ABA) stress. This can be attributed to the accumulation of Ph-glucanase at both transcript and protein levels during the seed germination in transgenic Arabidopsis. Results confirm that the cloned novel seed specific glucanase from a cold desert plant Podophyllum could be used for the manipulation of different plant species seeds against various harsh conditions. PMID:25303872

  20. [Spatial distribution of Tenuipalpus heveae Baker (Acari: Tenuipalpidae) on rubber tree plantations].

    PubMed

    Martins, Gustavo L M; Vieira, Marineide R; Barbosa, José C; Dini, Thiago A; Manzano, Anderson M; Alves, Bruno M S; Silva, Rodolfo M

    2010-01-01

    The objective of this work was to study the spatial distribution of Tenuipalpus heveae Baker in rubber tree plantations. The experimental area was located in Marinópolis, State of São Paulo, and corresponded to a total of 1,000 plants (clone RRIM 600) divided in 100 plots of ten plants each. A total of 16 samplings were conducted, approximately once every 10 days, between December 2007 and June 2008. On each date, samples were taken from two plants per plot, each sample corresponding to the top 30 cm of a branch randomly taken from the median region of the canopy of each plant. The number of T. heveae was evaluated on three leaflets randomly taken from each sample, using a 20x power pocket magnifying glass. The number of mites was evaluated in two areas of 1 cm² delimited on the lower surface of each leaflet, being one along the midrib and the other along a lateral vein. The calculated dispersion indexes were: variance/mean relationship (I), index of Morisita (I´), coefficient of Green (Cx) and k exponent of negative binomial distribution. Tenuipalpus heveae showed aggregate distribution. The negative binomial distribution model was the most appropriate to represent the spatial distribution of the mite in the rubber tree plantation. PMID:21120376

  1. Molecular Cloning, Expression Analysis, and Functional Characterization of the H(+)-Pyrophosphatase from Jatropha curcas.

    PubMed

    Yang, Yumei; Luo, Zhu; Zhang, Mengru; Liu, Chang; Gong, Ming; Zou, Zhurong

    2016-04-01

    H(+)-pyrophosphatase (H(+)-PPase) is a primary pyrophosphate (PPi)-energized proton pump to generate electrochemical H(+) gradient for ATP production and substance translocations across membranes. It plays an important role in stress adaptation that was intensively substantiated by numerous transgenic plants overexpressing H(+)-PPases yet devoid of any correlated studies pointing to the elite energy plant, Jatropha curcas. Herein, we cloned the full length of J. curcas H(+)-PPase (JcVP1) complementary DNA (cDNA) by reverse transcription PCR, based on the assembled sequence of its ESTs highly matched to Hevea brasiliensis H(+)-PPase. This gene encodes a polypeptide of 765 amino acids that was predicted as a K(+)-dependent H(+)-PPase evolutionarily closest to those of other Euphorbiaceae plants. Many cis-regulatory elements relevant to environmental stresses, molecular signals, or tissue-specificity were identified by promoter prediction within the 1.5-kb region upstream of JcVP1 coding sequence. Meanwhile, the responses of JcVP1 expression to several common abiotic stresses (salt, drought, heat, cold) were characterized with a considerable accordance with the inherent stress tolerance of J. curcas. Moreover, we found that the heterologous expression of JcVP1 could significantly improve the salt tolerance in both recombinant Escherichia coli and Saccharomyces cerevisiae, and this effect could be further fortified in yeast by N-terminal addition of a vacuole-targeting signal peptide from the H(+)-PPase of Trypanosoma cruzi. PMID:26643082

  2. Transcriptome analyses reveal molecular mechanism underlying tapping panel dryness of rubber tree (Hevea brasiliensis)

    PubMed Central

    Li, Dejun; Wang, Xuncheng; Deng, Zhi; Liu, Hui; Yang, Hong; He, Guangming

    2016-01-01

    Tapping panel dryness (TPD) is a serious threat to natural rubber yields from rubber trees, but the molecular mechanisms underlying TPD remain poorly understood. To identify TPD-related genes and reveal these molecular mechanisms, we sequenced and compared the transcriptomes of bark between healthy and TPD trees. In total, 57,760 assembled genes were obtained and analyzed in details. In contrast to healthy rubber trees, 5652 and 2485 genes were up- or downregulated, respectively, in TPD trees. The TPD-related genes were significantly enriched in eight GO terms and five KEGG pathways and were closely associated with ROS metabolism, programmed cell death and rubber biosynthesis. Our results suggest that rubber tree TPD is a complex process involving many genes. The observed lower rubber yield from TPD trees might result from lower isopentenyl diphosphate (IPP) available for rubber biosynthesis and from downregulation of the genes in post-IPP steps of rubber biosynthesis pathway. Our results not only extend our understanding of the complex molecular events involved in TPD but also will be useful for developing effective measures to control TPD of rubber trees. PMID:27005401

  3. A simple framework to analyze water constraints on seasonal transpiration in rubber tree (Hevea brasiliensis) plantations

    PubMed Central

    Sopharat, Jessada; Gay, Frederic; Thaler, Philippe; Sdoodee, Sayan; Isarangkool Na Ayutthaya, Supat; Tanavud, Charlchai; Hammecker, Claude; Do, Frederic C.

    2015-01-01

    Climate change and fast extension in climatically suboptimal areas threaten the sustainability of rubber tree cultivation. A simple framework based on reduction factors of potential transpiration was tested to evaluate the water constraints on seasonal transpiration in tropical sub-humid climates, according pedoclimatic conditions. We selected a representative, mature stand in a drought-prone area. Tree transpiration, evaporative demand and soil water availability were measured every day over 15 months. The results showed that basic relationships with evaporative demand, leaf area index and soil water availability were globally supported. However, the implementation of a regulation of transpiration at high evaporative demand whatever soil water availability was necessary to avoid large overestimates of transpiration. The details of regulation were confirmed by the analysis of canopy conductance response to vapor pressure deficit. The final objective of providing hierarchy between the main regulation factors of seasonal and annual transpiration was achieved. In the tested environmental conditions, the impact of atmospheric drought appeared larger importance than soil drought contrary to expectations. Our results support the interest in simple models to provide a first diagnosis of water constraints on transpiration with limited data, and to help decision making toward more sustainable rubber plantations. PMID:25610443

  4. Soil Carbon Stocks Decrease following Conversion of Secondary Forests to Rubber (Hevea brasiliensis) Plantations

    PubMed Central

    de Blécourt, Marleen; Brumme, Rainer; Xu, Jianchu; Corre, Marife D.; Veldkamp, Edzo

    2013-01-01

    Forest-to-rubber plantation conversion is an important land-use change in the tropical region, for which the impacts on soil carbon stocks have hardly been studied. In montane mainland southeast Asia, monoculture rubber plantations cover 1.5 million ha and the conversion from secondary forests to rubber plantations is predicted to cause a fourfold expansion by 2050. Our study, conducted in southern Yunnan province, China, aimed to quantify the changes in soil carbon stocks following the conversion from secondary forests to rubber plantations. We sampled 11 rubber plantations ranging in age from 5 to 46 years and seven secondary forest plots using a space-for-time substitution approach. We found that forest-to-rubber plantation conversion resulted in losses of soil carbon stocks by an average of 37.4±4.7 (SE) Mg C ha−1 in the entire 1.2-m depth over a time period of 46 years, which was equal to 19.3±2.7% of the initial soil carbon stocks in the secondary forests. This decline in soil carbon stocks was much larger than differences between published aboveground carbon stocks of rubber plantations and secondary forests, which range from a loss of 18 Mg C ha−1 to an increase of 8 Mg C ha−1. In the topsoil, carbon stocks declined exponentially with years since deforestation and reached a steady state at around 20 years. Although the IPCC tier 1 method assumes that soil carbon changes from forest-to-rubber plantation conversions are zero, our findings show that they need to be included to avoid errors in estimating overall ecosystem carbon fluxes. PMID:23894456

  5. Soil carbon stocks decrease following conversion of secondary forests to rubber (Hevea brasiliensis) plantations.

    PubMed

    de Blécourt, Marleen; Brumme, Rainer; Xu, Jianchu; Corre, Marife D; Veldkamp, Edzo

    2013-01-01

    Forest-to-rubber plantation conversion is an important land-use change in the tropical region, for which the impacts on soil carbon stocks have hardly been studied. In montane mainland southeast Asia, monoculture rubber plantations cover 1.5 million ha and the conversion from secondary forests to rubber plantations is predicted to cause a fourfold expansion by 2050. Our study, conducted in southern Yunnan province, China, aimed to quantify the changes in soil carbon stocks following the conversion from secondary forests to rubber plantations. We sampled 11 rubber plantations ranging in age from 5 to 46 years and seven secondary forest plots using a space-for-time substitution approach. We found that forest-to-rubber plantation conversion resulted in losses of soil carbon stocks by an average of 37.4±4.7 (SE) Mg C ha(-1) in the entire 1.2-m depth over a time period of 46 years, which was equal to 19.3±2.7% of the initial soil carbon stocks in the secondary forests. This decline in soil carbon stocks was much larger than differences between published aboveground carbon stocks of rubber plantations and secondary forests, which range from a loss of 18 Mg C ha(-1) to an increase of 8 Mg C ha(-1). In the topsoil, carbon stocks declined exponentially with years since deforestation and reached a steady state at around 20 years. Although the IPCC tier 1 method assumes that soil carbon changes from forest-to-rubber plantation conversions are zero, our findings show that they need to be included to avoid errors in estimating overall ecosystem carbon fluxes. PMID:23894456

  6. Physiological and molecular responses to drought stress in rubber tree (Hevea brasiliensis Muell. Arg.).

    PubMed

    Wang, Li-feng

    2014-10-01

    Plant drought stress response and tolerance are complex biological processes. In order to reveal the drought tolerance mechanism in rubber tree, physiological responses and expressions of genes involved in energy biosynthesis and reactive oxygen species (ROS) scavenging were systematically analyzed following drought stress treatment. Results showed that relative water content (RWC) in leaves was continuously decreased with the severity of drought stress. Wilting leaves were observed at 7 day without water (dww). Total chlorophyll content was increased at 1 dww, but decreased from 3 dww. However, the contents of malondialdehyde (MDA) and proline were significantly increased under drought stress. Peroxidase (POD) and superoxide dismutase (SOD) activities were markedly enhanced at 1 and 3 dww, respectively. Meanwhile, the soluble sugar content was constant under drought stress. These indicated that photosynthetic activity and membrane lipid integrity were quickly attenuated by drought stress in rubber tree, and osmoregulation participated in drought tolerance mechanism in rubber tree. Expressions of energy biosynthesis and ROS scavenging systems related genes, including HbCuZnSOD, HbMnSOD, HbAPX, HbCAT, HbCOA, HbATP, and HbACAT demonstrated that these genes were significantly up-regulated by drought stress, and reached a maximum at 3 dww, then followed by a decrease from 5 dww. These results suggested that drought stress adaption in rubber tree was governed by energy biosynthesis, antioxidative enzymes, and osmoregulation. PMID:25194774

  7. Genome-wide identification and characterization of WRKY gene family in Hevea brasiliensis.

    PubMed

    Li, Hui-Liang; Guo, Dong; Yang, Zi-Ping; Tang, Xiao; Peng, Shi-Qing

    2014-07-01

    WRKY proteins constitute a large family of transcription factors. In this study, we identified 81 WRKY genes (named HbWRKY1 to HbWRKY81) in the latest rubber tree genome. Tissue-specific expression profiles showed that 74 HbWRKYs were expressed in at least one of the tissues and the other 7 genes showed very low expression in all tissues tested, which suggested that HbWRKYs took part in many cellular processes. The responses of 20 selected HbWRKYs to jasmonic acid (JA) and ethylene (ET) were analyzed in the latex. 17 HbWRKYs responded to at least one treatment, which included 15 HbWRKYs responding to JA treatment, 15 HbWRKYs to ET, which suggested that these HbWRKYs were regulated by JA and ET. We also observed that HbWRKY3, 14, and 55 bind HbSRPP promoter and activate the transcription in yeast. This study suggests that HbWRKY proteins maybe involved in the transcriptional regulation of nature rubber biosynthesis. PMID:24793160

  8. Transcriptome analyses reveal molecular mechanism underlying tapping panel dryness of rubber tree (Hevea brasiliensis).

    PubMed

    Li, Dejun; Wang, Xuncheng; Deng, Zhi; Liu, Hui; Yang, Hong; He, Guangming

    2016-01-01

    Tapping panel dryness (TPD) is a serious threat to natural rubber yields from rubber trees, but the molecular mechanisms underlying TPD remain poorly understood. To identify TPD-related genes and reveal these molecular mechanisms, we sequenced and compared the transcriptomes of bark between healthy and TPD trees. In total, 57,760 assembled genes were obtained and analyzed in details. In contrast to healthy rubber trees, 5652 and 2485 genes were up- or downregulated, respectively, in TPD trees. The TPD-related genes were significantly enriched in eight GO terms and five KEGG pathways and were closely associated with ROS metabolism, programmed cell death and rubber biosynthesis. Our results suggest that rubber tree TPD is a complex process involving many genes. The observed lower rubber yield from TPD trees might result from lower isopentenyl diphosphate (IPP) available for rubber biosynthesis and from downregulation of the genes in post-IPP steps of rubber biosynthesis pathway. Our results not only extend our understanding of the complex molecular events involved in TPD but also will be useful for developing effective measures to control TPD of rubber trees. PMID:27005401

  9. Microdistribution of chromated copper arsenate preservative in rubberwood (Hevea brasiliensis Muell. Arg.)

    NASA Astrophysics Data System (ADS)

    Jusoh, Ismail Bin

    2000-08-01

    Rubberwood is popular for making indoor furniture since rubberwood is relatively abundant and sustainable. Currently more than 60% of the total annual rubberwood produced by rubber plantation is used as fuelwood. Rubberwood has the potential for both indoor and outdoor application. For exterior applications, preservative treatment is needed to extend the service life of rubberwood. The objectives of this study are to (1) assess treatability of rubberwood with chromated copper arsenate (CCA) preservative, (2) evaluate the natural decay resistance and efficacy of CCA on rubberwood, and (3) study the microdistribution of CCA components in rubberwood cells. The treatability of rubberwood was determined by measuring the penetration and retention of CCA type C preservative after a full-cell treatment. Natural decay resistance and efficacy of CCA treatment on rubberwood was estimated using a laboratory soilblock test according to AWPA E 10-91. The microdistribution of chromium, copper and arsenic in CCA-treated rubberwood was studied using scanning electron microscope in conjunction with energy dispersive X-ray analyzer (SEM-EDXA). As expected, longitudinal permeability was found to be better than the radial and the tangential permeability. The penetration and retention in the radial direction was about 3 times better than in the tangential direction. Longer pressure period increased penetration and retention of CCA type C in rubberwood. Complete penetration was achieved after 4 hours of pressure (1240 kPa) treatment. A pre-treatment steaming improved the treatability of rubberwood regardless of the anatomical direction. The average weight loss by white rot and brown rot was about 1.5 times higher than that of soft rot. A linear relationship was found between the weight loss and the incubation period for all the six test fungi. A CCA retention of 4.1 kg/m3 reduced weight loss to about 10% and retention of 14.5 kg/m3 reduced the weight loss of all test fungi at less than 2%. Vessels contained high level of chromium, copper, and arsenic compared to fibers. Chromium level was the highest, followed by arsenic and then copper in rubberwood cells. After the full cell treatment, fibers contained about 0.42%, 0.63%, and 1.02% of copper after treatment with 4.1 kg/m3, 10.5 kg/m 3, and 14.5 kg/m3 of CCA, respectively. Highest levels of Cr, Cu, and As were recorded in fiber-to-vessel cell corner (FVCC ) and Fiber-to-vessel middle lamella (FVML) and the lowest was recorded in S2 layer of fiber. Linescan analyses showed that higher count rates of carbon, oxygen, chromium, copper, and arsenic were found in the middle lamella compared to the fiber S2 layer in CCA-treated rubberwood. The increase of the solution strength in chromium, copper, and arsenic corresponds to an increase in Cr, Cu, and As level in wood cells.

  10. Histochemical and immunohistochemical identification of laticifer cells in callus cultures derived from anthers of Hevea brasiliensis.

    PubMed

    Tan, Deguan; Sun, Xuepiao; Zhang, Jiaming

    2011-06-01

    Laticifers are highly specialized cells present in over 20 plant families. They are well defined in planta. In vitro development of laticifers was also observed in some plants, but uncertain in the callus cultures of rubber tree, one of the most economically important latex producing plants. In the present study, we provide evidence that laticifer cells present in the callus cultures of rubber tree by histochemical and immunohistochemical studies. They present in the callus mainly as separate non-elongated form, a novel morphology different from the morphology of laticifer cells in planta, excluding their origin from explants. The occurring frequency of laticifer cells in the callus was genotype-dependent and negatively correlated with the somatic embryogenetic ability, suggesting that the presence of laticifer cells in the callus inhibit somatic embryogenesis in tissue culture of rubber tree. The genotypes PR107, RRIM600, Reyan8-79, and Reyan7-33-97 with lower embryogenetic ability compared to Haiken 2 had more laticifer cells, and laticifer clusters were only observed in these genotypes. PMID:21301851

  11. Phylogenetic Analysis Reveals a High Prevalence of Sporothrix brasiliensis in Feline Sporotrichosis Outbreaks

    PubMed Central

    Rodrigues, Anderson Messias; de Melo Teixeira, Marcus; de Hoog, G. Sybren; Schubach, Tânia Maria Pacheco; Pereira, Sandro Antonio; Fernandes, Geisa Ferreira; Bezerra, Leila Maria Lopes; Felipe, Maria Sueli; de Camargo, Zoilo Pires

    2013-01-01

    Sporothrix schenckii, previously assumed to be the sole agent of human and animal sporotrichosis, is in fact a species complex. Recently recognized taxa include S. brasiliensis, S. globosa, S. mexicana, and S. luriei, in addition to S. schenckii sensu stricto. Over the last decades, large epidemics of sporotrichosis occurred in Brazil due to zoonotic transmission, and cats were pointed out as key susceptible hosts. In order to understand the eco-epidemiology of feline sporotrichosis and its role in human sporotrichosis a survey was conducted among symptomatic cats. Prevalence and phylogenetic relationships among feline Sporothrix species were investigated by reconstructing their phylogenetic origin using the calmodulin (CAL) and the translation elongation factor-1 alpha (EF1α) loci in strains originated from Rio de Janeiro (RJ, n = 15), Rio Grande do Sul (RS, n = 10), Paraná (PR, n = 4), São Paulo (SP, n = 3) and Minas Gerais (MG, n = 1). Our results showed that S. brasiliensis is highly prevalent among cats (96.9%) with sporotrichosis, while S. schenckii was identified only once. The genotype of Sporothrix from cats was found identical to S. brasiliensis from human sources confirming that the disease is transmitted by cats. Sporothrix brasiliensis presented low genetic diversity compared to its sister taxon S. schenckii. No evidence of recombination in S. brasiliensis was found by split decomposition or PHI-test analysis, suggesting that S. brasiliensis is a clonal species. Strains recovered in states SP, MG and PR share the genotype of the RJ outbreak, different from the RS clone. The occurrence of separate genotypes among strains indicated that the Brazilian S. brasiliensis epidemic has at least two distinct sources. We suggest that cats represent a major host and the main source of cat and human S. brasiliensis infections in Brazil. PMID:23818999

  12. Sequential sampling plan for Tenuipalpus heveae Baker (Acari: Tenuipalpidae) on rubber tree.

    PubMed

    Martins, G L M; Vieira, M R; Barbosa, J C

    2013-04-01

    The objective of the present study was to develop a sequential sampling plan for the decision-making process to control Tenuipalpus heveae Baker (Acari: Tenuipalpidae), an important pest of the rubber tree crop. The experimental area was represented by 1,000 plants of the RRIM 600 clone divided in 100 plots with 10 plants each. Leaves were collected and the number of mites determined under laboratory conditions. The sequential sampling plan was developed in accordance with the Sequential Test Likelihood Ratio. The value 0.10 was pre-established for α and β representing type I and type II errors, respectively. The level of control adopted was six mites per 12 cm(2). The operating characteristic curve and the curve of maximum expected sample were determined. Two lines were generated: the upper one, when the condition for chemical control is recommended (S1 = 23.3080 + 2.1972); and the lower, when chemical control is not recommended (S0 = -23.3080 + 2.1972). Sample size for the decision-making process to control T. heveae requires 6 to 18 plants. PMID:23949755

  13. Elemental Analysis and Protein Quantification of Raw Natural Rubber of IAC Series 400 Clones

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Protein, nitrogen and sulfur contents were investigated for natural rubber from commercial Hevea, synthetic polyisoprene, and new IAC clones from Mococa city (IAC 405, 406, 410, 413, and 420), Jaú city (IAC 400, 401, 402, and 417), and from RRIM 600 clone (used as a control in both cases). IAC 405 a...

  14. Cytotaxonomy of the Brasiliensis subcomplex and the Triatoma brasiliensis complex (Hemiptera: Reduviidae: Triatominae).

    PubMed

    Alevi, Kaio C C; Rosa, João A; Azeredo-Oliveira, Maria Tercília V

    2014-01-01

    We analyzed the classical cytotaxonomy of the Brasiliensis subcomplex (Triatoma brasiliensis Neiva, T. juazeirensis Costa & Felix, T. melanica Costa, Argolo & Felix, T. melanocephala Neiva & Pinto, T. petrochiae Pinto & Barreto, T. lenti Sherlock & Serafim, T. sherlocki Papa, Jurberg, Carcavallo, Cerqueira & Barata, T. tibiamaculata Pinto and T. vitticeps Stal) and the T. brasiliensis complex (T. b. brasiliensis, T. b. macromelasoma Neiva & Lent, T. juazeirensis, T. melanica and T. sherlocki). The five members of the T. brasiliensis complex share the same cytogenetic characteristics. Merely T. sherlocki show differences in spermatids, which confirms the status of more differentiated member of the complex. T. lenti also presented the same cytogenetic characteristics described for the species of the T. brasiliensis complex, which supports possible grouping of the species as sixth member of the complex, although further analysis as molecular and experimental crosses are needed to corroborate this hypothesis. T. petrochiae, T. vitticeps, T. tibiamaculata and T. melanocephala presented one or more characteristics that allow questioning grouping in the proposed Brasiliensis subcomplex. Thus, we suggested that Brasiliensis subcomplex and T. brasiliensis complex should be constituted by the same triatomines (T. b. brasiliensis, T. b. macromelasoma, T. juazeirensis, T. melanica and T. sherlocki). However, we draw attention to T. lenti and suggest that although new analyzes should be performed, possibly this species is the sixth member of the T. brasiliensis complex.  PMID:25081800

  15. Nocardia brasiliensis: mycetoma induction and growth cycle.

    PubMed Central

    Conde, C; Melendro, E I; Fresán, M; Ortiz-Ortiz, L

    1982-01-01

    The capacity of Nocardia brasiliensis to induce mycetoma in BALB/c mice in the absence of adjuvants was studied. Whether the virulence of N. brasiliensis varied in the different phases of its growth cycle was also determined. The results showed that N. brasiliensis suspended in 0.15 M NaCl and injected into the footpads of mice were able to induce mycetoma after only 14 days of infection, as evidenced by histological studies. Data are also presented indicating that the virulence of N. brasiliensis did not vary during the different phases of its growth curve. The differences in virulence reported between N. brasiliensis and other nocardiae are discussed and explained in terms of several variables in the experimental designs used: among these variables are mouse genotype, route of inoculation, and model for determining virulence. Images PMID:7152671

  16. Identification of Paracoccidioides brasiliensis by gold nanoprobes

    NASA Astrophysics Data System (ADS)

    Martins, Jaciara F. S.; Castilho, Maiara L.; Cardoso, Maria A. G.; Carreiro, Andrea P.; Martin, Airton A.; Raniero, Leandro

    2012-01-01

    Paracoccidioides brasiliensis (P. brasiliensis) is a thermal dimorphic fungus and causal agent of paracoccidioidomycosis. Epidemiological data shows that it is mainly concentrated in Central and South America countries, with most registered cases in Colombia, Brazil, and Venezuela. The histopathological similarity with others fungal infection makes the diagnosis of P. brasiliensis more complicated. Therefore, the aim of this work was to find a positive and negative test for P. brasiliensis using gold nanoprobes as a new tool for P. brasiliensis detection. Gold nanoparticles were synthesized by reduction of gold chloride with sodium citrate. The results of this procedure is a wine-red solution with a maximum absorption in the range of ~520-530nm. A specific P. brasiliensis sequence of oligonucleotide was bonded to the nanoparticles, which maintained the wine-red color. The color changes from red to blue for negative diagnostic and is unchanged for a positive test. The H-bond interaction of DNA with the complementary DNA keeps strands together and forms double helical structure, maintaining the colloid stability. However, for non-complimentary DNA sequence the nanoprobes merge into a cluster, changing the light absorption.

  17. Optimisation on pretreatment of rubber seed (Hevea brasiliensis) oil via esterification reaction in a hydrodynamic cavitation reactor.

    PubMed

    Bokhari, Awais; Chuah, Lai Fatt; Yusup, Suzana; Klemeš, Jiří Jaromír; Kamil, Ruzaimah Nik M

    2016-01-01

    Pretreatment of the high free fatty acid rubber seed oil (RSO) via esterification reaction has been investigated by using a pilot scale hydrodynamic cavitation (HC) reactor. Four newly designed orifice plate geometries are studied. Cavities are induced by assisted double diaphragm pump in the range of 1-3.5 bar inlet pressure. An optimised plate with 21 holes of 1mm diameter and inlet pressure of 3 bar resulted in RSO acid value reduction from 72.36 to 2.64 mg KOH/g within 30 min of reaction time. Reaction parameters have been optimised by using response surface methodology and found as methanol to oil ratio of 6:1, catalyst concentration of 8 wt%, reaction time of 30 min and reaction temperature of 55°C. The reaction time and esterified efficiency of HC was three fold shorter and four fold higher than mechanical stirring. This makes the HC process more environmental friendly. PMID:26298387

  18. Characteristics of environmental Paracoccidioides brasiliensis isolates.

    PubMed

    Costa, Patricia Fagundes; Fernandes, Geisa Ferreira; dos Santos, Priscila Oliveira; Amaral, Cristiane Candida; Camargo, Zoilo Pires

    2010-01-01

    The ecological niche or exact habitat of the fungus Paracoccidioides brasiliensis is not known, and few isolates have been obtained from the environment. In this study, ten isolates were analyzed with respect to antigenic composition, serology, pathogenicity, and molecular aspects. Gp43 is considered to be the molecular basis for the serodiagnosis of paracoccidioidomycosis; however, in this study only six of the environmental isolates secreted this molecule (four in great amounts and two in small amounts). Other molecules were also produced. When exoantigens from these isolates were tested using immunodiffusion, only four preparations were positive by ID tests. However, when these exoantigens were tested by ELISA, all of them except one were able to detect anti-P. brasiliensis antibodies. In Western blot assays, these exoantigens showed different reactivities. Isolates that secreted gp43 presented positive reactions for this molecule, and isolates that did not secrete gp43 gave positive reactions for other minor molecules. RAPD analysis revealed that there is great genetic variation between these environmental isolates. These isolates were non-pathogenic: no mortality was observed among the inoculated mice during an 18-month follow-up period. PMID:19653119

  19. Potent Schistosomicidal Constituents from Garcinia brasiliensis.

    PubMed

    Castro, Aline Pereira; de Mattos, Ana Carolina Alves; Pereira, Neusa Araújo; Anchieta, Naira Ferreira; Silva, Matheus Siqueira; Dias, Danielle Ferreira; Silva, Claudinei Alves; Barros, Giulliano Vilela; Souza, Raquel Lopes Martins; Dos Santos, Marcelo Henrique; Marques, Marcos José

    2015-06-01

    Praziquantel is the drug of choice for the treatment of schistosomiasis. However, several strains of Schistosoma mansoni are resistant to praziquantel, making it necessary to discover new drugs that might be used for its treatment. With this in mind, the properties of a schistosomicidal ethanolic extract of Garcinia brasiliensis Mart. epicarp, the fractions obtained by partitioning this extract, including the hexane fractions, ethyl acetate fraction, and the aqueous fraction, and the isolated compounds 7-epiclusianone, a major component from these fractions, and fukugetin were tested in vitro on adult worms of S. mansoni. Mortality, damage to membranes, and excretory system activity were observed at 100.0, 50.0, 75.0, and 14.0 µg/mL for the ethanolic extract of G. brasiliensis Mart. epicarp, its hexane fractions, the ethyl acetate fraction, and 7-epiclusianone, respectively. For 7-epiclusianone, these data were confirmed by fluorescent probe Hoechst 33 258 and resorufin. Additionally, the biocidal effect of 7-epiclusianone was even higher than the hexane fractions. Moreover, an inhibitory effect of 7-epiclusianone on the egg laying of female adult S. mansoni worms was observed in cercariae and schistossomula. Thus, 7-epiclusianone is a promising schistosomicidal compound; however, more studies are needed to elucidate its mechanism of toxicity and to evaluate the in vivo activity of this compound. PMID:25905590

  20. Involvement of an Alternative Oxidase in Oxidative Stress and Mycelium-to-Yeast Differentiation in Paracoccidioides brasiliensis ▿ †

    PubMed Central

    Martins, Vicente P.; Dinamarco, Taisa M.; Soriani, Frederico M.; Tudella, Valéria G.; Oliveira, Sergio C.; Goldman, Gustavo H.; Curti, Carlos; Uyemura, Sérgio A.

    2011-01-01

    Paracoccidioides brasiliensis is a thermodimorphic human pathogenic fungus that causes paracoccidioidomycosis (PCM), which is the most prevalent systemic mycosis in Latin America. Differentiation from the mycelial to the yeast form (M-to-Y) is an essential step for the establishment of PCM. We evaluated the involvement of mitochondria and intracellular oxidative stress in M-to-Y differentiation. M-to-Y transition was delayed by the inhibition of mitochondrial complexes III and IV or alternative oxidase (AOX) and was blocked by the association of AOX with complex III or IV inhibitors. The expression of P. brasiliensis aox (Pbaox) was developmentally regulated through M-to-Y differentiation, wherein the highest levels were achieved in the first 24 h and during the yeast exponential growth phase; Pbaox was upregulated by oxidative stress. Pbaox was cloned, and its heterologous expression conferred cyanide-resistant respiration in Saccharomyces cerevisiae and Escherichia coli and reduced oxidative stress in S. cerevisiae cells. These results reinforce the role of PbAOX in intracellular redox balancing and demonstrate its involvement, as well as that of other components of the mitochondrial respiratory chain complexes, in the early stages of the M-to-Y differentiation of P. brasiliensis. PMID:21183691

  1. Protein influences on guayule and Hevea natural rubber sol and gel

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Guayule (Parthenium argentatum) is under cultivation in the southwestern United States as an alternative source of natural rubber free from proteins that cause Type I latex allergies. However, since guayule lacks the protein-polymer interactions present in Hevea latex, its physical and chemical prop...

  2. ASTM D1076 CATEGORY 4 LATEX AND QUANTIFYING GUAYULE (NRG) AND HEVEA (NR) LATEX PROTEIN

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Guayule latex is commercially available as a low protein natural rubber latex (Yulex®) which does not contain any protein that can be detected by the ASTM D6499 antigenic protein standard developed to quantify Hevea natural rubber latex (NRL) antigenic protein. In this paper, we discuss how best to...

  3. Immunohistochemical detection of a novel 22- to 25-kilodalton glycoprotein of Paracoccidioides brasiliensis in biopsy material and partial characterization by using species-specific monoclonal antibodies.

    PubMed Central

    Figueroa, J I; Hamilton, A; Allen, M; Hay, R

    1994-01-01

    Two murine monoclonal antibodies (MAbs) specific to Paracoccidioides brasiliensis (as determined by enzyme-linked immunosorbent assay [ELISA] and Western blot [immunoblot]) were produced by using a modification of standard hybridization protocols, with cyclophosphamide included as an immunomodulator to abolish responses to highly cross-reactive immunodominant epitopes. MAbs PS14 and PS15 are two different clones which exhibit similar characteristics by ELISA and Western blot. They are directed against a 22- to 25-kDa antigen which is present in P. brasiliensis and which could not be identified in other dimorphic fungi by ELISA or Western blot. Partial purification of the antigen was accomplished by isoelectric focusing, and deglycosylation studies suggested that the 22- to 25-kDa antigen is a glycoprotein with a pI of between 4.5 and 5 and that O-linked sugars may be part of the recognized epitope. The MAbs stained the cytoplasm of P. brasiliensis yeast and hyphal cells in cryostat sections of fresh cultures of the fungus. In addition, the MAbs stained the wall of paracoccidioidomycotic granulomas, as well as the cytoplasm of the fungus, as determined by the use of immunofluorescence, immunoperoxidase, and immuno-alkaline phosphatase staining techniques in paraffin-embedded sections of human biopsy material, and they failed to stain granulomas resulting from other clinical conditions. These findings suggest that these MAbs have potential use in the immunohistochemical identification of P. brasiliensis. Images PMID:8077405

  4. Why Clone?

    MedlinePlus

    ... How might cloning be used in medicine? Cloning animal models of disease Much of what researchers learn about human disease comes from studying animal models such as mice. Often, animal models are ...

  5. [Microclimatic properties of the Triatoma brasiliensis habitat].

    PubMed

    Lorenzo, M G; Guarneri, A A; Pires, H H; Diotaiuti, L; Lazzari, C R

    2000-01-01

    Vector-borne transmission of Chagas disease in Northeast Brazil is basically by Triatoma brasiliensis. It is thus crucial to determine this species' microclimatic preferences as limiting factors for its distribution and ability to infest domestic environments. We analyze the microclimatic properties of the shelters in which these insects are found in wild, domestic, and peridomiciliary environments in the State of Ceará, at Brazil. We measure temperature and relative humidity (RH) every 15 minutes for 3 days. Thermal variation was greatly dampened inside both domiciliary refuges and the more protected internal places in wild stony sites. For RH, we observed a similar dampening pattern, but mean RH was lower in both domiciliary refuges and wild ones inside stony sites as compared to reference levels in the surrounding environment. The results are discussed with regard to this species' microclimatic preferences in the laboratory and its potential as determinants of its geographical distribution. PMID:11119321

  6. cDNA encoding a polypeptide including a hevein sequence

    DOEpatents

    Raikhel, N.V.; Broekaert, W.F.; Namhai Chua; Kush, A.

    1993-02-16

    A cDNA clone (HEV1) encoding hevein was isolated via polymerase chain reaction (PCR) using mixed oligonucleotides corresponding to two regions of hevein as primers and a Hevea brasiliensis latex cDNA library as a template. HEV1 is 1,018 nucleotides long and includes an open reading frame of 204 amino acids.

  7. Ultrastructure of Dimorphic Transformation in Paracoccidioides brasiliensis

    PubMed Central

    Carbonell, Luis M.

    1969-01-01

    The fine structure of Paracoccidioides brasiliensis undergoing temperature-dependent transformation from mycelium to yeast and vice versa (M ⇌ Y) was studied. The transitional form to mycelium from the yeast appears as an elongated bud that extends from the yeast and which has a mixture of characteristics from both the yeast and the mycelium. The transitional form to yeast from the mycelium starts with enlargement of the interseptal spaces and cracking of the outer electron-dense layer of the cell wall of the hypha. Later the interseptal spaces tend to become round and separate. In M → Y only few interseptal spaces seem to transform. The yeast is produced by self-transformation of the hypha. In Y → M a new structure is formed and the yeast dies. Intrahyphal hyphae are observed during the transformation from M → Y, and intrayeast hyphae during the Y → M. Due to the high mortality and breakage observed in both types of transformations, we believe that wound of the yeast or the mycelium could elicit this phenomenon. Images PMID:5359610

  8. Characterization of the cellular antigens of Paracoccidioides brasiliensis yeast form.

    PubMed Central

    Casotto, M

    1990-01-01

    Antigenic components of the yeast extract of Paracoccidioides brasiliensis Linder 2511 cultured for 3, 8, 20, 30, and 60 days were examined by the Western blot (immunoblot) technique. The 3-day extract was chosen for characterization of the antigenic components because its stability did not vary with time and it contained all antigens identified by patient sera. Antibodies to cross-reacting antigens of P. brasiliensis extracts were detected in sera from patients with histoplasmosis, candidiasis, and aspergillosis. The 58-, 57-, 21-, and 16-kilodalton (kDa) antigens were specific for P. brasiliensis, while the 48- and 45-kDa antigens were specific for paracoccidioidomycosis. The Western blot technique is a useful tool for the diagnosis of disease and revealed heterogeneity in the responses of patient sera. The combination of the 58-, 57-, and 45-kDa proteins confirmed a diagnosis of paracoccidioidomycosis (87% of the cases). Images PMID:2380351

  9. Academic Cloning.

    ERIC Educational Resources Information Center

    Sikula, John P.; Sikula, Andrew F.

    1980-01-01

    The authors define "cloning" as an integral feature of all educational systems, citing teaching practices which reward students for closely reproducing the teacher's thoughts and/or behaviors and administrative systems which tend to promote like-minded subordinates. They insist, however, that "academic cloning" is not a totally negative practice.…

  10. Monoclonal antibodies to Nocardia asteroides and Nocardia brasiliensis antigens.

    PubMed Central

    Jiménez, T; Díaz, A M; Zlotnik, H

    1990-01-01

    Nocardia asteroides and Nocardia brasiliensis whole-cell extracts were used as antigens to generate monoclonal antibodies (MAbs). Six stable hybrid cell lines secreting anti-Nocardia spp. MAbs were obtained. These were characterized by enzyme-linked immunosorbent assay, Western blot (immunoblot), and immunofluorescence assay. Although all the MAbs exhibited different degrees of cross-reactivity with N. asteroides and N. brasiliensis antigens as well as with culture-filtrate antigens from Mycobacteria spp., they have the potential for use as reagents in the purification of Nocardia antigens. Images PMID:2405017

  11. Kalanchosine dimalate, an anti-inflammatory salt from Kalanchoe brasiliensis.

    PubMed

    Costa, Sônia Soares; de Souza, Maria de Lourdes Mendes; Ibrahim, Tereza; de Melo, Giany Oliveira; de Almeida, Ana Paula; Guette, Catherine; Férézou, Jean-Pierre; Koatz, Vera Lucia G

    2006-05-01

    This report describes the isolation and characterization of kalanchosine dimalate (KMC), an anti-inflammatory salt from the fresh juice of the aerial parts of Kalanchoe brasiliensis. KMC comprises the new metabolite kalanchosine (1) and malic acid (2) in a 1:2 stoichiometric ratio. Kalanchosine (1), 3,6-diamino-4,5-dihydroxyoctanedioic acid, is the first naturally occurring dimeric bis(gamma-hydroxy-beta-amino acid) and is at least partially responsible for the anti-inflammatory properties of K. brasiliensis. PMID:16724848

  12. Larva of Palaemnema brasiliensis Machado (Odonata: Platystictidae), from Amazonas, Brazil.

    PubMed

    Neiss, Ulisses Gaspar; Hamada, Neusa

    2016-01-01

    The larva of Palaemnema brasiliensis Machado, 2009 is described and illustrated based on last-instar larvae and exuviae of reared larvae collected in a blackwater stream in Barcelos and Presidente Figueiredo municipalities, Amazonas state, Brazil. The larva of P. brasiliensis can be distinguished from the two South American species of the genus with described larvae (P. clementia Selys and P. mutans Calvert), mainly by presence of a single obtuse cusp on the labial palp, the presence and configuration of setae in the caudal lamellae, and the proportional length of terminal filaments of the caudal lamellae. The family is recorded here for the first time in Brazilian state of Amazonas. PMID:27395963

  13. Effects of methanol-to-oil ratio, catalyst amount and reaction time on the FAME yield by in situ transesterification of rubber seeds (Hevea brasiliensis)

    NASA Astrophysics Data System (ADS)

    Abdulkadir, Bashir Abubakar; Uemura, Yoshimitsu; Ramli, Anita; Osman, Noridah B.; Kusakabe, Katsuki; Kai, Takami

    2014-10-01

    In this research, biodiesel is produced by in situ transesterification (direct transesterification) method from the rubber seeds using KOH as a catalyst. The influence of methanol to seeds mass ratio, duration of reaction, and catalyst loading was investigated. The result shows that, the best ratio of seeds to methanol is 1:6 (10 g seeds with 60 g methanol), 120 minutes reaction time and catalyst loading of 3.0 g. The maximum FAME yield obtain was 70 %. This findings support FAME production from the seeds of rubber tree using direct transesterifcation method from the seeds of rubber tree as an alternative to diesel fuel. Also, significant properties of biodiesel such as cloud point, density, pour point, specific gravity, and viscosity were investigated.

  14. ANIMAL MODEL OF NIPPOSTRONGYLUS BRASILIENSIS AND HELIGMOSOMOIDES POLYGYRUS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Animal models of Nippostrongylus brasiliensis and Heligmosomoides polygyrus infection are powerful tools for the investigation of the basic biology of immune responses and protective immunity. In particular they model the induction and maintenance of Th2 type immune responses and exhibiting all the ...

  15. Temporal dynamics of reproduction in Hemiramphus brasiliensis (Osteichthyes: Hemiramphidae).

    PubMed

    de Oliveira, Mônica Rocha; Chellappa, Sathyabama

    2014-01-01

    The reproductive aspects of Hemiramphus brasiliensis were analyzed with a view to verify the temporal dynamics of reproduction. This paper presents data on sex ratio, length at first sexual maturity, macroscopic and histological aspects of gonad development, gonadosomatic index (GSI), reproductive period, and fecundity of H. brasiliensis. The fishes were captured from the coastal waters of Rio Grande do Norte, northeastern Brazil. Females of this species predominated in the sampled population and were larger in size than the males. The length at the first sexual maturation of males was 20.8 cm and that of females was 21.5 cm. The macroscopic characteristics of the gonads indicated four maturation stages. Histological studies of gonads of H. brasiliensis showed six phases of oocyte development and four phases of spermatocyte development. The batch fecundity of this species was 1153 (±258.22) mature oocytes for 50 g body weight of female. The microscopic characteristics of gonad development indicate that H. brasiliensis is a multiple spawner, presenting a prolonged reproductive period during the whole year, with a peak in the month of April, and is considered as an opportunistic strategist. PMID:25512946

  16. 5-Lipoxygenase Activity Increases Susceptibility to Experimental Paracoccidioides brasiliensis Infection

    PubMed Central

    Tristão, Fabrine Sales Massafera; Rocha, Fernanda Agostini; Moreira, Ana Paula; Cunha, Fernando Queiroz; Rossi, Marcos Antonio

    2013-01-01

    Paracoccidioidomycosis (PCM) is a systemic mycosis caused by the thermodimorphic fungus Paracoccidioides brasiliensis. Leukotrienes and lipoxins are lipid mediators produced after 5-lipoxygenase (5-LO) activation that exhibit pro- and anti-inflammatory roles, respectively. Here, we have investigated the contribution of 5-LO enzymatic activity in PCM using an experimental model of P. brasiliensis infection. B6.129 wild-type (B6.129) and 5-LO-deficient (5-LO−/−) mice were intravenously inoculated with a virulent strain of P. brasiliensis (Pb18), and the survival rate of the infected mice was investigated on different days after yeast infection. 5-LO−/− mice exhibited an increased survival rate associated with a decreased number of CFU. The resistance of 5-LO−/− during PCM was associated with augmented nitric oxide (NO) production and the formation of compact granulomas. In addition, the absence of 5-LO was associated with a diminished number of CD4+ CD25+ regulatory T cells, higher levels of gamma interferon and interleukin-12, and increased T-bet (a T-box transcription factor that directs Th1 lineage commitment) mRNA levels in the lungs. Taken together, our results show for the first time that 5-LO enzymatic activity increases susceptibility to P. brasiliensis, suggesting that this pathway may be a potential target for therapeutic intervention during PCM. PMID:23381993

  17. Temporal Dynamics of Reproduction in Hemiramphus brasiliensis (Osteichthyes: Hemiramphidae)

    PubMed Central

    de Oliveira, Mônica Rocha

    2014-01-01

    The reproductive aspects of Hemiramphus brasiliensis were analyzed with a view to verify the temporal dynamics of reproduction. This paper presents data on sex ratio, length at first sexual maturity, macroscopic and histological aspects of gonad development, gonadosomatic index (GSI), reproductive period, and fecundity of H. brasiliensis. The fishes were captured from the coastal waters of Rio Grande do Norte, northeastern Brazil. Females of this species predominated in the sampled population and were larger in size than the males. The length at the first sexual maturation of males was 20.8 cm and that of females was 21.5 cm. The macroscopic characteristics of the gonads indicated four maturation stages. Histological studies of gonads of H. brasiliensis showed six phases of oocyte development and four phases of spermatocyte development. The batch fecundity of this species was 1153 (±258.22) mature oocytes for 50 g body weight of female. The microscopic characteristics of gonad development indicate that H. brasiliensis is a multiple spawner, presenting a prolonged reproductive period during the whole year, with a peak in the month of April, and is considered as an opportunistic strategist. PMID:25512946

  18. Paracoccidioides brasiliensis infection in domestic rabbits (Oryctolagus cuniculus).

    PubMed

    Belitardo, Donizeti Rodrigues; Calefi, Atilio Sersun; Sbeghen, Mônica Raquel; de Oliveira, Gabriela Gonçalves; Watanabe, Maria Angelica Ehara; de Camargo, Zoilo Pires; Ono, Mario Augusto

    2014-04-01

    The objective of this study was to evaluate the infection of domestic rabbits by Paracoccidioides brasiliensis. Initially two rabbits were experimentally infected with P. brasiliensis and the humoral immune response was evaluated by ELISA using gp43 as antigen. The two animals showed IgG response against gp43 although no signs of disease were observed. The seroepidemiological study was carried out in 170 rabbits (free range n = 81 and caged n = 89) living in an endemic area for human paracoccidioidomycosis and a positivity of 27% was observed in the ELISA using gp43 as antigen. The free-range rabbits showed a significantly higher positivity (34.6-51.7%) than the caged animals (11.1%). Sentinel rabbits exposed to natural infection with P. brasiliensis were followed up for 6 months and a seroconversion rate of 83.3% was observed. This is the first report of paracoccidioidomycosis in rabbits and suggests that this species can be useful sentinels for P. brasiliensis presence in the environment. PMID:24125519

  19. Recombinant Paracoccin Reproduces the Biological Properties of the Native Protein and Induces Protective Th1 Immunity against Paracoccidioides brasiliensis Infection

    PubMed Central

    Alegre, Ana Claudia Paiva; Oliveira, Aline Ferreira; Dos Reis Almeida, Fausto Bruno; Roque-Barreira, Maria Cristina; Hanna, Ebert Seixas

    2014-01-01

    Background Paracoccin is a dual-function protein of the yeast Paracoccidioides brasiliensis that has lectin properties and N-acetylglucosaminidase activities. Proteomic analysis of a paracoccin preparation from P. brasiliensis revealed that the sequence matched that of the hypothetical protein encoded by PADG-3347 of isolate Pb-18, with a polypeptide sequence similar to the family 18 endochitinases. These endochitinases are multi-functional proteins, with distinct lectin and enzymatic domains. Methodology/principal findings The multi-exon assembly and the largest exon of the predicted ORF (PADG-3347), was cloned and expressed in Escherichia coli cells, and the features of the recombinant proteins were compared to those of the native paracoccin. The multi-exon protein was also used for protection assays in a mouse model of paracoccidioidomycosis. Conclusions/Significance Our results showed that the recombinant protein reproduced the biological properties described for the native protein—including binding to laminin in a manner that is dependent on carbohydrate recognition—showed N-acetylglucosaminidase activity, and stimulated murine peritoneal macrophages to produce high levels of TNF-α and nitric oxide. Considering the immunomodulatory potential of glycan-binding proteins, we also investigated whether prophylactic administration of recombinant paracoccin affected the course of experimental paracoccidioidomycosis in mice. In comparison to animals injected with vehicle (controls), mice treated with recombinant paracoccin displayed lower pulmonary fungal burdens and reduced pulmonary granulomas. These protective effects were associated with augmented pulmonary levels of IL-12 and IFN-γ. We also observed that injection of paracoccin three days before challenge was the most efficient administration protocol, as the induced Th1 immunity was balanced by high levels of pulmonary IL-10, which may prevent the tissue damage caused by exacerbated inflammation. The

  20. Paracoccidioides brasiliensis AND Paracoccidioides lutzii, A SECRET LOVE AFFAIR.

    PubMed

    Arantes, Thales Domingos; Bagagli, Eduardo; Niño-Vega, Gustavo; San-Blas, Gioconda; Theodoro, Raquel Cordeiro

    2015-09-01

    To commemorate Prof. Carlos da Silva Lacaz's centennial anniversary, the authors have written a brief account of a few, out of hundreds, biological, ecological, molecular and phylogenetic studies that led to the arrival of Paracoccidioides lutzii, hidden for more than a century within Paracoccidioides brasiliensis. Lacaz's permanent interest in this fungus, and particularly his conviction on the benefits that research on paracoccidioidomycosis would bring to patients, were pivotal in the development of the field. PMID:26465366

  1. Extracellular Paracoccidioides brasiliensis phospholipase B involvement in alveolar macrophage interaction

    PubMed Central

    2010-01-01

    Background Phospholipase B (PLB) has been reported to be one of the virulence factors for human pathogenic fungi and has also been described as necessary for the early events in infection. Based on these data, we investigated the role of PLB in virulence and modulation of the alveolar pulmonary immune response during infection using an in-vitro model of host-pathogen interaction, i.e. Paracoccidioides brasiliensis yeast cells infecting alveolar macrophage (MH-S) cells. Results The effect of PLB was analyzed using the specific inhibitor alexidine dihydrochloride (0.25 μM), and pulmonary surfactant (100 μg mL-1), during 6 hours of co-cultivation of P. brasiliensis and MH-S cells. Alexidine dihydrochloride inhibited PLB activity by 66% and significantly decreased the adhesion and internalization of yeast cells by MH-S cells. Genes involved in phagocytosis (trl2, cd14) and the inflammatory response (nfkb, tnf-α, il-1β) were down-regulated in the presence of this PLB inhibitor. In contrast, PLB activity and internalization of yeast cells significantly increased in the presence of pulmonary surfactant; under this condition, genes such as clec2 and the pro-inflammatory inhibitor (nkrf) were up-regulated. Also, the pulmonary surfactant did not alter cytokine production, while alexidine dihydrochloride decreased the levels of interleukin-10 (IL-10) and increased the levels of IL-12 and tumor necrosis factor-α (TNF-α). In addition, gene expression analysis of plb1, sod3 and icl1 suggests that P. brasiliensis gene re-programming is effective in facilitating adaptation to this inhospitable environment, which mimics the lung-environment interaction. Conclusion P. brasiliensis PLB activity is involved in the process of adhesion and internalization of yeast cells at the MH-S cell surface and may enhance virulence and subsequent down-regulation of macrophage activation. PMID:20843362

  2. Paracoccidioides brasiliensis AND Paracoccidioides lutzii, A SECRET LOVE AFFAIR

    PubMed Central

    ARANTES, Thales Domingos; BAGAGLI, Eduardo; NIÑO-VEGA, Gustavo; SAN-BLAS, Gioconda; THEODORO, Raquel Cordeiro

    2015-01-01

    SUMMARY To commemorate Prof. Carlos da Silva Lacaz's centennial anniversary, the authors have written a brief account of a few, out of hundreds, biological, ecological, molecular and phylogenetic studies that led to the arrival of Paracoccidioides lutzii, hidden for more than a century within Paracoccidioides brasiliensis. Lacaz's permanent interest in this fungus, and particularly his conviction on the benefits that research on paracoccidioidomycosis would bring to patients, were pivotal in the development of the field. PMID:26465366

  3. Ecotopes, natural infection and trophic resources of Triatoma brasiliensis (Hemiptera, Reduviidae, Triatominae).

    PubMed

    Costa, J; de Almeida, J R; Britto, C; Duarte, R; Marchon-Silva, V; Pacheco, R da S

    1998-01-01

    Triatoma brasiliensis is considered as one of the most important Chagas disease vectors in the northeastern Brazil. This species presents chromatic variations which led to descriptions of subspecies, synonymized by Lent and Wygodzinsky (1979). In order to broaden bionomic knowledge of these distinct colour patterns of T. brasiliensis, captures were performed at different sites, where the chromatic patterns were described: Caicó, Rio Grande do Norte (T. brasiliensis brasiliensis Neiva, 1911), it will be called the "brasiliensis population"; Espinosa, Minas Gerais (T. brasiliensis melanica Neiva & Lent 1941), the "melanica population" and Petrolina, Pernambuco (T. brasiliensis macromelasoma, Galvão 1956), the "macromelasoma population". A fourth chromatic pattern was collected in Juazeiro, Bahia the darker one in overall cuticle coloration, the "Juazeiro population". At the sites of Caicó, Petrolina and Juazeiro, specimens were captured in peridomiciliar ecotopes and in wilderness. In Espinosa the specimens were collected only in wilderness, even though several exhaustive captures have been performed in peridomicile at different sites of this municipality. A total of 298 specimens were captured. The average registered infection rate was 15% for "brasiliensis population" and of 6.6% for "melanica population". Specimens of "macromelasoma" and of "Juazeiro populations" did not present natural infection. Concerning trophic resources, evaluated by the precipitin test, feeding eclecticism for the different colour patterns studied was observed, with dominance of goat blood in household surroundings as well as in wilderness. PMID:9698835

  4. Revalidation and redescription of Triatoma brasiliensis macromelasoma Galvão, 1956 and an identification key for the Triatoma brasiliensis complex (Hemiptera: Reduviidae: Triatominae)

    PubMed Central

    Costa, Jane; Correia, Nathália Cordeiro; Neiva, Vanessa Lima; Gonçalves, Teresa Cristina Monte; Felix, Márcio

    2013-01-01

    Triatoma brasiliensis macromelasoma is revalidated based on the results of previous multidisciplinary studies on the Triatoma brasiliensis complex, consisting of crossing experiments and morphological, biological, ecological and molecular analyses. These taxonomic tools showed the closest relationship between T. b. macromelasoma and Triatoma brasiliensis brasiliensis. T. b. macromelasoma is redescribed based on specimens collected in the type locality and specimens from a F1 colony. The complex now comprises T. b. brasiliensis, T. b. macromelasoma, Triatoma melanica, Triatoma juazeirensis and Triatoma sherlocki. An identification key for all members of the complex is presented. This detailed comparative study of the morphological features of T. b. macromelasoma and the remaining members of the complex corroborates results from multidisciplinary analyses, suggesting that the subspecific status is applicable. This subspecies can be distinguished by the following combination of features: a pronotum with 1+1 narrow brownish-yellow stripes on the submedian carinae, not attaining its apex, hemelytra with membrane cells darkened on the central portion and legs with an incomplete brownish-yellow ring on the apical half of the femora. Because the T. brasiliensis complex is of distinct epidemiological importance throughout its geographic distribution, a precise identification of its five members is important for monitoring and controlling actions against Chagas disease transmission. PMID:24037202

  5. In vitro susceptibility testing of Paracoccidioides brasiliensis to sulfonamides.

    PubMed Central

    Restrepo, A; Arango, M D

    1980-01-01

    A total of 60 clinical isolates of Paracoccidioides brasiliensis were tested for susceptibility to sulfadiazine and sulfadimethoxyne by the agar dilution technique. A modification of the Mueller-Hinton medium was devised which gave good growth of the yeast form. The minimum inhibitory concentrations for only 51.6% of the isolates were in the range of the recommended blood serum concentration (50 micrograms/ml). For 6 to 8% of the isolates, the minimum inhibitory concentrations were above 200 micrograms of both sulfadiazine and sulfadimethoxyne per ml. A significant decreases in susceptibility was demonstrated for one isolate obtained from a patient relapsing during sulfonamide therapy. Images PMID:7416744

  6. Molecular cloning and expression of a novel MYB transcription factor gene in rubber tree.

    PubMed

    Qin, Bi; Zhang, Yu; Wang, Meng

    2014-12-01

    MYB family proteins regulate a variety of cellular processes in plants. Tapping panel dryness (TPD) in rubber tree (Hevea brasiliensis Muell. Arg.) affects latex biosynthesis and causes serious losses to rubber producers. In this study, a novel SANT/MYB transcription factor gene down-regulated in TPD rubber tree, named as HbSM1, was isolated from rubber tree. The complete HbSM1 open reading frame (ORF) was 948 bp in length. The deduced HbSM1 protein is 315 amino acids. HbSM1 belonged to 1RMYB subfamily with a single SANT domain. Sequence alignment revealed that HbSM1 had high homology with MYB members from Ricinus communis and Manihot esculenta, with 72 and 78 % identity, respectively. Moreover, HbSM1 shared 56 % identity with Glycine max GmMYB176. Phylogenetic analysis revealed that HbSM1, GmMYB176, rice OsMYBS2, and OsMYBS3 fell into the same cluster with 93 % bootstrap support value. Comparing expression among different tissues demonstrated that HbSM1 was ubiquitously expressed in all tissues, but it appeared to be preferentially expressed in leaf and latex. Furthermore, HbSM1 transcripts were significantly induced by various phytohormones (including gibberellic acid, ethephon, methyl jasmonate, salicylic acid, and abscisic acid) and wounding treatments. These results suggested that HbSM1 might play multiple roles in plant development via different phytohormones signaling pathways. PMID:25195053

  7. Isolation and characterization of microsatellite markers in the Serra Spanish mackerel, Scomberomorus brasiliensis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Thirteen nuclear-encoded microsatellites from a genomic DNA library of Serra Spanish mackerel, Scomberomorus brasiliensis, were isolated and characterized. The microsatellites include 10 perfect repeats (8 tetranucleotide and 2 dinucleotide) and 3 imperfect repeats (2 tetranucleotide and 1 dinucleo...

  8. New records of pyrrolizidine alkaloid-feeding insects. Hemiptera and Coleoptera on Senecio brasiliensis.

    PubMed

    Klitzke; Trigo

    2000-04-01

    New records are reported for a Hemipteran (Largus rufipennis) and a Coleopteran (Chauliognathus fallax) feeding on Senecio brasiliensis (Asteraceae) and sequestering pyrrolizidine alkaloids (PAs). Both insects are warningly colored and rejected by predators. PMID:10725590

  9. Isolation and characterization of microsatellite markers in the Serra Spanish mackerel, Scomberomorus brasiliensis.

    PubMed

    Renshaw, Mark A; Douglas, Kory C; Rexroad Iii, Caird E; Jobity, Ann Marie C; Gold, John R

    2009-05-01

    Thirteen nuclear-encoded microsatellites from a genomic DNA library of Serra Spanish mackerel, Scomberomorus brasiliensis, were isolated and characterized. The microsatellites include 10 perfect repeats (eight tetranucleotide and two dinucleotide) and three imperfect repeats (two tetranucleotide and one dinucleotide). An additional five microsatellites, isolated originally from two congeneric species (S. cavalla and S. niphonius), were characterized in S. brasiliensis. Serra Spanish mackerel support artisanal fisheries along the Caribbean and Atlantic coasts of Central and South America, from Belize to Brazil. PMID:21564761

  10. In Vitro Activities of the New Antitubercular Agents PA-824 and BTZ043 against Nocardia brasiliensis

    PubMed Central

    Campos-Rivera, Mayra Paola; Gonzalez-Martinez, Norma Alejandra; Ocampo-Candiani, Jorge; Cole, Stewart T.

    2012-01-01

    The in vitro activity of PA-824 and BTZ043 against 30 Nocardia brasiliensis isolates was tested. The MIC50 and MIC90 values for PA-824 were both >64 μg/ml. The same values for BTZ043 were 0.125 and 0.250 μg/ml. Given the MIC values for benzothiazinone (BTZ) compounds, we consider them good candidates to be tested in vivo against N. brasiliensis. PMID:22526312

  11. In vitro activities of the new antitubercular agents PA-824 and BTZ043 against Nocardia brasiliensis.

    PubMed

    Vera-Cabrera, Lucio; Campos-Rivera, Mayra Paola; Gonzalez-Martinez, Norma Alejandra; Ocampo-Candiani, Jorge; Cole, Stewart T

    2012-07-01

    The in vitro activity of PA-824 and BTZ043 against 30 Nocardia brasiliensis isolates was tested. The MIC(50) and MIC(90) values for PA-824 were both >64 μg/ml. The same values for BTZ043 were 0.125 and 0.250 μg/ml. Given the MIC values for benzothiazinone (BTZ) compounds, we consider them good candidates to be tested in vivo against N. brasiliensis. PMID:22526312

  12. The Clone Factory

    ERIC Educational Resources Information Center

    Stoddard, Beryl

    2005-01-01

    Have humans been cloned? Is it possible? Immediate interest is sparked when students are asked these questions. In response to their curiosity, the clone factory activity was developed to help them understand the process of cloning. In this activity, students reenact the cloning process, in a very simplified simulation. After completing the…

  13. Complete mitochondrial genome of Salminus brasiliensis (Characiformes, Characidae).

    PubMed

    Brandão-Dias, Pedro Ferreira Pinto; Carmo, Anderson Oliveira do; Martins, Ana Paula Vimieiro; Pimenta, Ricardo José Gonzaga; Alves, Carlos Bernardo Mascarenhas; Kalapothakis, Evanguedes

    2016-05-01

    We report the complete mitochondrial genome of the fish Salminus brasiliensis, popularly known as dourado. It is a circular, 17,721 bp long DNA molecule, containing 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and a non-coding Control Region of 2128 bp, a relatively large molecule when compared to other closely related fishes. All protein-coding genes are on the heavy strand, except for Nd6, and all have ATG as the start codon, except for Cox1 gene which has a GTG start codon. Seven protein-coding genes have incomplete stop codons (Nd2, Cox2, Atp6, Nd4, and Cytb have T- -, and Cox3 and Nd3 have TA-). TAG is the stop codon for Nd6 and AGG is the stop codon for Cox1. PMID:25208163

  14. Drimanes from Drimys brasiliensis with leishmanicidal and antimalarial activity

    PubMed Central

    Claudino, Vanessa Duarte; da Silva, Kesia Caroline; Cechinel, Valdir; Yunes, Rosendo Augusto; Monache, Franco Delle; Giménez, Alberto; Salamanca, Efrain; Gutierrez-Yapu, David; Malheiros, Angela

    2013-01-01

    This paper evaluates CHCl3 and CH3OH extracts of the stem bark, branches and leaves of Drimys brasiliensis and drimane sesquiterpenes isolated from the stem bark against strains of Leishmania amazonensis and Leishmania braziliensis promastigotes and Plasmodium falciparum trophozoites. All of the extracts and compounds were tested in cell lines in comparison with reference standards and cell viability was determined by the XTT method. The CHCl3 and CH3OH extracts from the stem bark and branches yielded promising results against two strains of Leishmania, with 50% inhibitory concentrations (IC50 ) values ranging from 39-100 µg/mL. The CHCl3 extract of the stem bark returned IC50 values of 39 and 40.6 µg/mL for L. amazonensis and L. braziliensis, respectively. The drimanes were relatively effective: 1-β-(p-coumaroyloxy)-polygodial produced IC50 values of 5.55 and 2.52 µM for L. amazonensis and L. braziliensis, respectively, compared with 1-β-(p-methoxycinnamoyl)-polygodial, which produced respective IC50 values of 15.85 and 17.80 µM. The CHCl3 extract demonstrated activity (IC50 of 3.0 µg/mL) against P. falciparum. The IC50 values of 1-β-(p-cumaroyloxyl)-polygodial and 1-β-(p-methoxycinnamoyl)-polygodial were 1.01 and 4.87 µM, respectively, for the trophozoite strain. Therefore, the results suggest that D. brasiliensis is a promising plant from which to obtain new and effective antiparasitic agents. PMID:23579790

  15. Antimicrobial effect of farnesol, a Candida albicans quorum sensing molecule, on Paracoccidioides brasiliensis growth and morphogenesis

    PubMed Central

    Derengowski, Lorena S; De-Souza-Silva, Calliandra; Braz, Shélida V; Mello-De-Sousa, Thiago M; Báo, Sônia N; Kyaw, Cynthia M; Silva-Pereira, Ildinete

    2009-01-01

    Background Farnesol is a sesquiterpene alcohol produced by many organisms, and also found in several essential oils. Its role as a quorum sensing molecule and as a virulence factor of Candida albicans has been well described. Studies revealed that farnesol affect the growth of a number of bacteria and fungi, pointing to a potential role as an antimicrobial agent. Methods Growth assays of Paracoccidioides brasiliensis cells incubated in the presence of different concentrations of farnesol were performed by measuring the optical density of the cultures. The viability of fungal cells was determined by MTT assay and by counting the colony forming units, after each farnesol treatment. The effects of farnesol on P. brasiliensis dimorphism were also evaluated by optical microscopy. The ultrastructural morphology of farnesol-treated P. brasiliensis yeast cells was evaluated by transmission and scanning electron microscopy. Results In this study, the effects of farnesol on Paracoccidioides brasiliensis growth and dimorphism were described. Concentrations of this isoprenoid ranging from 25 to 300 μM strongly inhibited P. brasiliensis growth. We have estimated that the MIC of farnesol for P. brasiliensis is 25 μM, while the MLC is around 30 μM. When employing levels which don't compromise cell viability (5 to 15 μM), it was shown that farnesol also affected the morphogenesis of this fungus. We observed about 60% of inhibition in hyphal development following P. brasiliensis yeast cells treatment with 15 μM of farnesol for 48 h. At these farnesol concentrations we also observed a significant hyphal shortening. Electron microscopy experiments showed that, despite of a remaining intact cell wall, P. brasiliensis cells treated with farnesol concentrations above 25 μM exhibited a fully cytoplasmic degeneration. Conclusion Our data indicate that farnesol acts as a potent antimicrobial agent against P. brasiliensis. The fungicide activity of farnesol against this pathogen is

  16. Antagonistic Activity of Nocardia brasiliensis PTCC 1422 Against Isolated Enterobacteriaceae from Urinary Tract Infections.

    PubMed

    Jalali, Hossnieh Kafshdar; Salamatzadeh, Abdolreza; Jalali, Arezou Kafshdar; Kashani, Hamed Haddad; Asbchin, Salman Ahmadi; Issazadeh, Khosro

    2016-03-01

    The main drawback of current antibiotic therapies is the emergence and rapid increase in antibiotic resistance. Nocardiae are aerobic, Gram-positive, catalase-positive, non-motile actinomycetes. Nocardia brasiliensis was reported as antibiotic producer. The purpose of the study was to determine antibacterial activity of N. brasiliensis PTCC 1422 against isolated Enterobacteriaceae from urinary tract infections (UTIs). The common bacteria from UTIs were isolated from hospital samples. Antimicrobial susceptibility test was performed for the isolated pathogens using Kirby-Bauer disk diffusion method according to clinical and Laboratory Standards Institute guideline. Antagonistic activity of N. brasiliensis PTCC 1422 was examined with well diffusion methods. Supernatant of N. brasiliensis PTCC 1422 by submerged culture was analyzed with gas chromatography-mass spectrometry. Isolated strains included Escherichia coli, Klebsiella pneumoniae, Serratia marcescens and Proteus mirabilis. The most common pathogen isolated was E. coli (72.5%). Bacterial isolates revealed the presence of high levels of antimicrobial resistances to ceftriaxone and low levels of resistance to cephalexin. Supernatant of N. brasiliensis PTCC 1422 showed antibacterial activity against all of the isolated microorganisms in well diffusion method. The antibiotic resistance among the uropathogens is an evolving process, so a routine surveillance to monitor the etiologic agents of UTI and the resistance pattern should be carried out timely to choose the most effective empirical treatment by the physicians. Our present investigation indicates that the substances present in the N. brasiliensis PTCC 1422 could be used to inhibit the growth of human pathogen. Antibacterial resistance among bacterial uropathogen is an evolving process. Therefore, in the field on the need of re-evaluation of empirical treatment of UTIs, our present. The study has demonstrated that N. brasiliensis PTCC 1422 has a high potential

  17. Draft Genome Sequence of Pseudozyma brasiliensis sp. nov. Strain GHG001, a High Producer of Endo-1,4-Xylanase Isolated from an Insect Pest of Sugarcane

    PubMed Central

    Oliveira, Juliana Velasco de Castro; dos Santos, Renato Augusto Corrêa; Borges, Thuanny A.

    2013-01-01

    Here, we present the nuclear and mitochondrial genome sequences of Pseudozyma brasiliensis sp. nov. strain GHG001. P. brasiliensis sp. nov. is the closest relative of Pseudozyma vetiver. P. brasiliensis sp. nov. is capable of growing on xylose or xylan as a sole carbon source and has great biotechnological potential. PMID:24356824

  18. Draft Genome Sequence of Pseudozyma brasiliensis sp. nov. Strain GHG001, a High Producer of Endo-1,4-Xylanase Isolated from an Insect Pest of Sugarcane.

    PubMed

    Oliveira, Juliana Velasco de Castro; Dos Santos, Renato Augusto Corrêa; Borges, Thuanny A; Riaño-Pachón, Diego Mauricio; Goldman, Gustavo Henrique

    2013-01-01

    Here, we present the nuclear and mitochondrial genome sequences of Pseudozyma brasiliensis sp. nov. strain GHG001. P. brasiliensis sp. nov. is the closest relative of Pseudozyma vetiver. P. brasiliensis sp. nov. is capable of growing on xylose or xylan as a sole carbon source and has great biotechnological potential. PMID:24356824

  19. Multipartite asymmetric quantum cloning

    SciTech Connect

    Iblisdir, S.; Gisin, N.; Acin, A.; Cerf, N.J.; Filip, R.; Fiurasek, J.

    2005-10-15

    We investigate the optimal distribution of quantum information over multipartite systems in asymmetric settings. We introduce cloning transformations that take N identical replicas of a pure state in any dimension as input and yield a collection of clones with nonidentical fidelities. As an example, if the clones are partitioned into a set of M{sub A} clones with fidelity F{sup A} and another set of M{sub B} clones with fidelity F{sup B}, the trade-off between these fidelities is analyzed, and particular cases of optimal N{yields}M{sub A}+M{sub B} cloning machines are exhibited. We also present an optimal 1{yields}1+1+1 cloning machine, which is an example of a tripartite fully asymmetric cloner. Finally, it is shown how these cloning machines can be optically realized.

  20. Aristotle and headless clones.

    PubMed

    Mosteller, Timothy

    2005-01-01

    Cloned organisms can be genetically altered so that they do not exhibit higher brain functioning. This form of therapeutic cloning allows for genetically identical organs and tissues to be harvested from the clone for the use of the organism that is cloned. "Spare parts" cloning promises many opportunities for future medical advances. What is the ontological and ethical status of spare parts, headless clones? This paper attempts to answer this question from the perspective of Aristotle's view of the soul. Aristotle's metaphysics as applied to his view of biological essences generates an ethic that can contribute to moral reasoning regarding the use of headless spare parts clones. The task of this paper is to show the implications that Aristotle's view of the soul, if it is true, would have on the ethics of headless, spare parts cloning. PMID:16180113

  1. Exocellular components of Paracoccidioides brasiliensis: identification of a specific antigen.

    PubMed Central

    Puccia, R; Schenkman, S; Gorin, P A; Travassos, L R

    1986-01-01

    Yeast forms of Paracoccidioides brasiliensis grown in liquid medium produced exocellular components. Immunodiffusion reactions and immunoprecipitations of 131I-radiolabeled antigenic components with sera from patients having paracoccidioidomycosis (PCM) were used to monitor the isolation of specific constituents. Components having the main antigenic activity (fCon A) were isolated by exclusion from a Bio-Gel P30 column, followed by successive binding of eluted material to a Sepharose-concanavalin A column, and elution. The product contained, from sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, a minor 43,000-molecular-weight (MW) component (gp43), a polydisperse high-MW glycoconjugate, and a diffusely migrating 55,000-MW glycoprotein (gp55). Other components, including a 72,000-MW glycoprotein, were irregularly expressed. The high-MW glycoconjugate complex contained, on the basis of methylation and 13C nuclear magnetic resonance data, a branched structure of mainly mannopyranosyl units. These were nonreducing ends, 6-O-, 2-O-, and 2,6-di-O-substituted, and the specific rotation of +16 degrees indicated that the glycosidic configurations of the units were alpha and beta in a ratio of ca. 1:1 (concanavalin A binding indicated that nonreducing ends or 2-O-substituted units or both of alpha-D-mannopyranose were present). A small proportion of nonreducing end units of D-galactopyranose were also present in this polysaccharide. gp55 is a glycoprotein containing a complex carbohydrate moiety with fucose, mannose, galactose, and glucose, either as terminal nonreducing units or substituted in positions indicated by methylation data. Both PCM and normal human sera precipitated the high-MW glycoconjugate from 131I-labeled fCon A preparations, whereas gp55 was unreactive with human sera. gp43 was a specific antigenic component of P. brasiliensis culture filtrates which could be isolated in a pure form by gel filtration column chromatography (Sephadex G150

  2. Phenotypic variability confirmed by nuclear ribosomal DNA suggests a possible natural hybrid zone of Triatoma brasiliensis species complex.

    PubMed

    Costa, Jane; Bargues, Maria Dolores; Neiva, Vanessa Lima; Lawrence, Gena G; Gumiel, Marcia; Oliveira, Genova; Cabello, Pedro; Lima, Marli Maria; Dotson, Ellen; Provance, David William; Almeida, Carlos Eduardo; Mateo, Lucia; Mas-Coma, Santiago; Dujardin, Jean Pierre

    2016-01-01

    Triatoma brasiliensis macromelasoma occurs in Pernambuco state, Brazil, which is situated between the distribution areas of Triatoma brasiliensis brasiliensis (north) and Triatoma juazeirensis (south). T. b. macromelasoma displays greater variations in its chromatic phenotype than either T. b. brasiliensis or T. juazeirensis, and patterns reminiscent of one or the other. Experimental crosses from each of these members of the T. brasiliensis species complex generated fertile offspring suggesting that viable hybrids could be present in nature, despite their significant genetic distances. Considering the geographical position of occurrence of the T. b. macromelasoma (in Pernambuco) it was proposed to be an area capable of supporting natural hybridization between T. b. brasiliensis and T. juazeirensis. Since phenotypic variability is expected, this study investigated the existence of intermediate chromatic phenotypes for T. b. macromelasoma in various locations in areas between the T. b. brasiliensis and T. juazeirensis occurrences. Thirteen different color patterns were for the first time characterized and nine of those displayed intermediate phenotypes. Molecular analysis performed using ribosomal DNA intergenic region, grouped all within the T. brasiliensis complex. The intermediate chromatic phenotypes, molecular analysis and experimental crosses all support the distinction of a zone of hybridization that gave rise to the T. b. macromelasoma through homoploidal evolution. PMID:26520796

  3. Seroepidemiology of Paracoccidioides brasiliensis infection in horses from Rio Grande do Sul, Brazil.

    PubMed

    Albano, Ana Paula Neuschrank; Klafke, Gabriel Baracy; Brandolt, Tchana Martinez; Da Hora, Vanusa Pousada; Nogueira, Carlos Eduardo Wayne; Xavier, Melissa Orzechowski; Meireles, Mário Carlos Araújo

    2015-06-01

    Paracoccidioides brasiliensis is the etiological agent of the major systemic mycosis in Brazil, called paracoccidioidomycosis. Although the Rio Grande do Sul is considered an endemic area of the disease, there are few studies on the ecology of P. brasiliensis in the state. Therefore, this study aimed to evaluate the infection of P. brasiliensis in horses from the mesoregion of Southwest Riograndense, using these animals as sentinels. Serological techniques, such as double immunodiffusion in agar gel (AGID) and indirect ELISA, were performed to detect the anti-gp43 P. brasiliensis antibody in horses from five different farms in the region of Bagé, RS, Brazil. Serology was performed in 200 Pure Blood English horses up to two years of age that were born and raised exclusively at the farms. Of these horses, 12% had anti-gp43 antibodies according to the ELISA results, with rates ranging from 0 to 30% according to the farm of origin (p < 0.001). Based on the immunodiffusion results, all equine serum samples were negative. These results indicate the presence of the fungus P. brasiliensis in the middle region of the southwestern state of Rio Grande do Sul, Brazil. PMID:26273267

  4. Sensitive immunoradiometric assay for the detection of Paracoccidioides brasiliensis antigens in human sera.

    PubMed Central

    Ferreira-da-Cruz, M F; Galvão-Castro, B; Daniel-Ribeiro, C T

    1991-01-01

    In the present study we report the standardization of an immunoradiometric assay (IRMA) for detection of Paracoccidioides brasiliensis circulating antigens that could be useful in the diagnosis and prognosis of paracoccidioidomycosis. For this purpose we studied the reactivities of P. brasiliensis and other mycotic antigens with rabbit polyclonal anti-P. brasiliensis antibodies (immunoglobulin G) in order to evaluate the sensitivity and specificity of an IRMA for detecting P. brasiliensis antigens. The results were compared with those obtained by the double immunodiffusion test, the standard technique for the serodiagnosis of paracoccidioidomycosis. By using the immunoglobulin G fraction of rabbit antisera (900 ng per well), it was possible to detect up to 3.6 ng (0.12 micrograms/ml) of cellular antigen and 360 ng (12 micrograms/ml) of metabolic antigen in contrast to the double immunodiffusion test that could detect only 12 micrograms (1.2 mg/ml) of both antigens. IRMA was shown to be feasible and very sensitive and may therefore help, together with clinical data, in establishing early diagnosis and assessing disease activity. It could also allow the study of relationships between P. brasiliensis circulating antigens and host defense mechanisms during the disease. PMID:1907608

  5. Lymphocutaneous Nocardia brasiliensis infection acquired from a cat scratch: case report and review.

    PubMed

    Sachs, M K

    1992-10-01

    Nocardia brasiliensis is a bacterium that is most commonly found in the soil. Traumatic inoculation of N. brasiliensis into the skin is the most typical mode of acquisition of infection due to this organism. To the best of my knowledge, I report the first case of lymphocutaneous N. brasiliensis disease from a penetrating cat scratch of the skin, thereby establishing cats as vehicles for the transmission of this infection. Treatment with penicillin produced a rapid resolution of all signs and symptoms of infection. The efficacy of penicillin against the N. brasiliensis isolate recovered from this patient was highly unusual. In general, penicillin has limited therapeutic value since these organisms elaborate beta-lactamase. Sulfonamides remain the drugs of choice for the treatment of these infections. These soil-borne organisms are most likely carried on the claws of cats and may establish infection after percutaneous inoculation. A high index of suspicion for N. brasiliensis soft-tissue infection is required since a delayed or missed diagnosis may be associated with progressive local disease and/or widespread disseminated infection. PMID:1420688

  6. Natural products from Garcinia brasiliensis as Leishmania protease inhibitors.

    PubMed

    Pereira, Ivan O; Assis, Diego M; Juliano, Maria A; Cunha, Rodrigo L O R; Barbieri, Clara L; do Sacramento, Luis V S; Marques, Marcos J; dos Santos, Marcelo H

    2011-06-01

    The infections by protozoans of the genus Leishmania are a major worldwide health problem, with high endemicity in developing countries. The drugs of choice for the treatment of leishmaniasis are the pentavalent antimonials, which cause renal and cardiac toxicity. As part of a search for new drugs against leishmaniasis, we evaluated the in vitro Leishmania protease inhibition activity of extracts (hexanic, ethyl-acetate, and ethanolic) and fukugetin, a bioflavonoid purified from the ethyl-acetate extract of the pericarp of the fruit of Garcinia brasiliensis, a tree native to Brazilian forests. The isolated compound was characterized by using spectral analyses with nuclear magnetic resonance, mass spectroscopy, ultraviolet, and infrared techniques. The ethyl-acetate extract and the compound fukugetin showed significant activity as inhibitors of Leishmania's proteases, with mean (±SD) IC(50) (50% inhibition concentration of protease activity) values of 15.0±1.3 μg/mL and 3.2±0.5 μM/mL, respectively, characterizing a bioguided assay. In addition, this isolated compound showed no activity against promastigote and amastigote forms of L. (L.) amazonensis and mammalian cells. These results suggest that fukugetin is a potent protease inhibitor of L. (L.) amazonensis and does not cause toxicity in mammalian or Leishmania cells in vitro. This study provides new perspectives on the development of novel drugs that have leishmanicidal activity obtained from natural products and that target the parasite's proteases. PMID:21554130

  7. New iridoid glucosides from the aerial parts of Verbena brasiliensis.

    PubMed

    Ono, Masateru; Oishi, Kanna; Abe, Hiroaki; Masuoka, Chikako; Okawa, Masafumi; Ikeda, Tsuyoshi; Nohara, Toshihiro

    2006-10-01

    Two new iridoid glucosides, verbenabraside A (1) and verbenabraside B (2), were isolated from the aerial parts of Verbena brasiliensis VELL., along with six known iridoid glucosides, gelsemiol 3-O-beta-D-glucoside (3), verbraside (4), 9-hydroxysemperoside (5), griselinoside (6), aralidioside (7), and 6alpha-hydroxyforsythide dimethyl ester (8), three known phenylethanoid glycosides, 2-phenylethyl O-beta-D-xylopyranosyl-(1-->2)-beta-D-glucopyranoside (9), acteoside (10), and leucosceptoside A (11), two known lignan glucosides, dihydroxymethyl-bis(3,5-dimethoxy-4-hydroxyphenyl) tetrahydrofuran-9 (or 9')-O-beta-glucopyranoside (12) and (+)-lyoniresinol 3alpha-O-beta-D-glucopyranoside (13), a known methyl salicylate glucoside, methyl 2-O-beta-D-glucopyranosylbenzoate (14), and two known sterols, beta-sitosterol 3-O-beta-D-glucopyranoside (15) and beta-sitosterol (16). Their chemical structures were determined on the basis of spectroscopic data. Compound 1 exhibited stronger scavenging effect on the stable free radical 1,1-diphenyl-2-picrylhydrazyl than that of alpha-tocopherol. PMID:17015981

  8. Cryptic species of Paracoccidioides brasiliensis: impact on paracoccidioidomycosis immunodiagnosis

    PubMed Central

    Machado, Gabriel Capella; Moris, Daniela Vanessa; Arantes, Thales Domingos; Silva, Luciane Regina Franciscone; Theodoro, Raquel Cordeiro; Mendes, Rinaldo Pôncio; Vicentini, Adriana Pardini; Bagagli, Eduardo

    2013-01-01

    We aimed to evaluate whether the occurrence of cryptic species of Paracoccidioides brasiliensis, S1, PS2, PS3 and Paracoccidioides lutzii, has implications in the immunodiagnosis of paracoccidioidomycosis (PCM). Small quantities of the antigen gp43 were found in culture filtrates of P. lutzii strains and this molecule appeared to be more variable within P. lutzii because the synonymous-nonsynonymous mutation rate was lower, indicating an evolutionary process different from that of the remaining genotypes. The production of gp43 also varied between isolates belonging to the same species, indicating that speciation events are important, but not sufficient to fully explain the diversity in the production of this antigen. The culture filtrate antigen AgEpm83, which was obtained from a PS3 isolate, showed large quantities of gp43 and reactivity by immunodiffusion assays, similar to the standard antigen (AgB-339) from an S1 isolate. Furthermore, AgEpm83 was capable of serologically differentiating five serum samples from patients from the Botucatu and Jundiaí regions. These patients had confirmed PCM but, were non-reactive to the standard antigen, thus demonstrating an alternative for serological diagnosis in regions in which S1 and PS2 occur. We also emphasise that it is not advisable to use a single antigen preparation to diagnose PCM, a disease that is caused by highly diverse pathogens. PMID:23903981

  9. Reconstructing the locomotor repertoire of Protopithecus brasiliensis. I. Body size.

    PubMed

    Halenar, Lauren B

    2011-12-01

    An accurate body size estimate is essential for reconstructing and interpreting many aspects of the paleobiology of an extinct taxon. With this in mind, the purpose of this study is two-fold: first, to create statistically robust predictive regression equations for body mass, total body length, and head and body length from postcranial elements using a platyrrhine reference sample, data that do not exist elsewhere in the literature; and, second, to apply those regression equations to the "giant" subfossil platyrrhine Protopithecus brasiliensis, a little-studied taxon represented by a nearly complete skeleton. Building on results of previous work with other primate groups, different skeletal elements, subgroups of the reference sample, and regression models lead to different body size estimates with different standard errors and prediction errors. However, relatively tight clusters of estimates around 20 kg, total length of 1,675 mm, and head and body length of 710 mm are obtained, placing the fossil in the size range of a large male baboon. While not quite as large as the original 25 kg body mass estimate for the fossil, this new estimate is still approximately 150% larger than the largest living New World monkey. Confirmation of its place in a large-bodied size class of platyrrhines has a profound effect on reconstructing the locomotor repertoire of Protopithecus and the evolutionary trajectory of the alouattin lineage. PMID:22042663

  10. Sporothrix brasiliensis outbreaks and the rapid emergence of feline sporotrichosis.

    PubMed

    Sanchotene, Karine Ortiz; Madrid, Isabel Martins; Klafke, Gabriel Baracy; Bergamashi, Mariana; Della Terra, Paula Portella; Rodrigues, Anderson Messias; de Camargo, Zoilo Pires; Xavier, Melissa Orzechowski

    2015-11-01

    Sporotrichosis is the main subcutaneous mycosis in Brazil, and is caused by Sporothrix schenckii and allied species. Sporothrix propagules present on soil and plant debris may be traumatically inoculated into the cutaneous/ subcutaneous tissues of the warm-blooded host. An alternative route involves direct animal-animal and animal-human transmissions through deep scratches and bites of diseased cats. Sporotrichosis is much more common than previously appreciated with several cases emerging over the years especially in South and Southeast Brazil. We conducted an epidemiological surveillance in endemic areas of feline sporotrichosis in the southern region of Rio Grande do Sul state, Brazil. Over the last 5-year period the number of feline sporotrichosis in Rio Grande increased from 0.75 new cases per month in 2010 to 3.33 cases per month in 2014. The wide geographic distribution of diagnosed cases highlights the dynamics of Sporothrix transmission across urban areas with high population density. Molecular identification down to species level by PCR-RFLP of cat-transmitted Sporothrix revealed the emergence of the clonal offshoot S. brasiliensis during feline outbreaks; this scenario is similar to the epidemics taking place in the metropolitan areas of Rio de Janeiro and São Paulo. Controlling and preventing sporotrichosis outbreaks are essential steps to managing the disease among humans and animals. PMID:26404561

  11. Preparation of species-specific murine monoclonal antibodies against the yeast phase of Paracoccidioides brasiliensis.

    PubMed Central

    Figueroa, J I; Hamilton, A J; Bartholomew, M A; Harada, T; Fenelon, L; Hay, R J

    1990-01-01

    A panel of four murine monoclonal antibodies showing species specificity for the yeast phase of the pathogenic dimorphic fungus Paracoccidioides brasiliensis was produced by using a modification of the standard monoclonal antibody technology. This involved the use of the immunosuppressive drug cyclophosphamide to suppress the immune response of test animals to fungi showing cross-reactivity, i.e., to Histoplasma capsulatum. One monoclonal antibody, P4, which had a high titer by enzyme-linked immunosorbent assay, was shown to recognize a linear antigenic epitope of P. brasiliensis at a molecular size of 70,000 to 75,000 daltons by Western blot (immunoblot) analysis. The potential use of these monoclonal antibodies, which are the first species-specific probes to P. brasiliensis that have been produced, in the field of serodiagnosis is discussed. Images PMID:2394802

  12. Increase in virulence of Sporothrix brasiliensis over five years in a patient with chronic disseminated sporotrichosis

    PubMed Central

    Freitas, Dayvison FS; Santos, Suelen S; Almeida-Paes, Rodrigo; de Oliveira, Manoel ME; do Valle, Antonio CF; Gutierrez-Galhardo, Maria Clara; Zancopé-Oliveira, Rosely M; Nosanchuk, Joshua d

    2015-01-01

    Abstract The metropolitan region of Rio de Janeiro is hyperendemic for cat-associated sporotrichosis. This study aimed to assess the virulence of serial Sporothrix isolates from a 61-year-old male patient with chronic, destructive disseminated sporotrichosis. Five Sporothrix isolates were cultured from skin exudates and bone samples over a 5-year period, and all were molecularly identified as Sporothrix brasiliensis. The final isolate was significantly more virulent in Galleria mellonella larvae compared to earlier isolates. We conclude that S. brasiliensis has the capacity to increase in virulence in vivo. This finding is significant to clinicians caring for individuals with S. brasiliensis disease and it suggests that further studies are needed to identify the mechanisms underlying pathogenicity enhancement during chronic disease. PMID:25668479

  13. Diversity and antimicrobial potential of culturable heterotrophic bacteria associated with the endemic marine sponge Arenosclera brasiliensis

    PubMed Central

    Rua, Cintia P.J.; Trindade-Silva, Amaro E.; Appolinario, Luciana R.; Venas, Tainá M.; Garcia, Gizele D.; Carvalho, Lucas S.; Lima, Alinne; Kruger, Ricardo; Pereira, Renato C.; Berlinck, Roberto G.S.; Valle, Rogério A.B.; Thompson, Cristiane C.

    2014-01-01

    Marine sponges are the oldest Metazoa, very often presenting a complex microbial consortium. Such is the case of the marine sponge Arenosclera brasiliensis, endemic to Rio de Janeiro State, Brazil. In this investigation we characterized the diversity of some of the culturable heterotrophic bacteria living in association with A. brasiliensis and determined their antimicrobial activity. The genera Endozoicomonas (N = 32), Bacillus (N = 26), Shewanella (N = 17), Pseudovibrio (N = 12), and Ruegeria (N = 8) were dominant among the recovered isolates, corresponding to 97% of all isolates. Approximately one third of the isolates living in association with A. brasiliensis produced antibiotics that inhibited the growth of Bacillus subtilis, suggesting that bacteria associated with this sponge play a role in its health. PMID:25024903

  14. Reproductive Biology of Triatoma brasiliensis (Hemiptera, Triatominae) During the Imaginal Molt.

    PubMed

    Alevi, Kaio Cesar Chaboli; Guerra, Ana Letícia; Imperador, Carlos Henrique Lima; da Rosa, João Aristeu; de Azeredo-Oliveira, Maria Tercília Vilela

    2016-03-01

    The triatomines are vectors of the protozoan Trypanosoma cruzi, etiologic agent of Chagas disease. These insects are sexually active after the imaginal molt. Some aspects have been studied in Triatoma brasiliensis during the imaginal molt, such as autogeny in virgin females and the relationship between blood ingestion by fifth instar nymph and the realization of the imaginal molt. Thus, to aid in the understanding of reproductive biology and developmental physiology of these vectors, this article analyzes the spermatogenesis of T. brasiliensis during the imaginal molt. The analysis of the seminiferous tubules from males in the fifth instar during imaginal molt has demonstrated that T. brasiliensis has only a few spermatids and a plentiful quantity of sperm. Thus, we suggest that during imaginal molt the cell division is disrupted aiming to reduce energy costs and the differentiation into sperm is stimulated to ensure the paternity of the adult male. PMID:26787143

  15. Proteomic Analysis Reveals That Iron Availability Alters the Metabolic Status of the Pathogenic Fungus Paracoccidioides brasiliensis

    PubMed Central

    Parente, Ana F. A.; Bailão, Alexandre M.; Borges, Clayton L.; Parente, Juliana A.; Magalhães, Adriana D.; Ricart, Carlos A. O.; Soares, Célia M. A.

    2011-01-01

    Paracoccidioides brasiliensis is a thermodimorphic fungus and the causative agent of paracoccidioidomycosis (PCM). The ability of P. brasiliensis to uptake nutrients is fundamental for growth, but a reduction in the availability of iron and other nutrients is a host defense mechanism many pathogenic fungi must overcome. Thus, fungal mechanisms that scavenge iron from host may contribute to P. brasiliensis virulence. In order to better understand how P. brasiliensis adapts to iron starvation in the host we compared the two-dimensional (2D) gel protein profile of yeast cells during iron starvation to that of iron rich condition. Protein spots were selected for comparative analysis based on the protein staining intensity as determined by image analysis. A total of 1752 protein spots were selected for comparison, and a total of 274 out of the 1752 protein spots were determined to have changed significantly in abundance due to iron depletion. Ninety six of the 274 proteins were grouped into the following functional categories; energy, metabolism, cell rescue, virulence, cell cycle, protein synthesis, protein fate, transcription, cellular communication, and cell fate. A correlation between protein and transcript levels was also discovered using quantitative RT-PCR analysis from RNA obtained from P. brasiliensis under iron restricting conditions and from yeast cells isolated from infected mouse spleens. In addition, western blot analysis and enzyme activity assays validated the differential regulation of proteins identified by 2-D gel analysis. We observed an increase in glycolytic pathway protein regulation while tricarboxylic acid cycle, glyoxylate and methylcitrate cycles, and electron transport chain proteins decreased in abundance under iron limiting conditions. These data suggest a remodeling of P. brasiliensis metabolism by prioritizing iron independent pathways. PMID:21829521

  16. Conservation genetics of the giant otter (Pteronura brasiliensis (Zimmerman, 1780)) (Carnivora, Mustelidae).

    PubMed

    Garcia, D M; Marmontel, M; Rosas, F W; Santos, F R

    2007-12-01

    The giant otter (Pteronura brasiliensis) is an aquatic mammal of the Mustelidae family, endemic to South America. Its original distribution corresponds to the region from the Guyanas to Central-North Argentina, but it is extinct or on the verge of extinction in most of its historical range. Currently, the species is considered endangered by the World Conservation Union (IUCN). Based on its geographic distribution in the South American continent and on some morphological characters, two subspecies were suggested: P. brasiliensis brasiliensis, occurring in the Amazon and Orinoco River Basins, and P. brasiliensis paranensis, in the Paraná and Paraguai River Basins. However, there is no consensus on assuming this subspecies division and no detailed studies have been carried out to elucidate this question. This study aims to evaluate the genetic diversity and population structure of Pteronura brasiliensis along its range in Brazil to check the possibility of the existence of two distinct subspecies using also a reciprocal monophyly criterion. We analyzed the control region, and the Cytochrome b and Cytochrome c Oxidase subunit I genes of the mitochondrial DNA in several giant otter populations from the Amazon and Paraguai River Basins. Analyses have indicated some degree of geographic correlation and a high level of inter-population divergence, although the subspecies division is not highly supported. As we observed strong population structure, we cannot rule out the existence of further divisions shaping the species distribution. The results suggest that a more complex population structure occurs in P. brasiliensis, and the conservation practice should concentrate on preserving all remaining local populations. PMID:18278348

  17. TLR9 Activation Dampens the Early Inflammatory Response to Paracoccidioides brasiliensis, Impacting Host Survival

    PubMed Central

    Menino, João Filipe; Saraiva, Margarida; Gomes-Alves, Ana G.; Lobo-Silva, Diogo; Sturme, Mark; Gomes-Rezende, Jéssica; Saraiva, Ana Laura; Goldman, Gustavo H.; Cunha, Cristina; Carvalho, Agostinho; Romani, Luigina; Pedrosa, Jorge; Castro, António Gil; Rodrigues, Fernando

    2013-01-01

    Background Paracoccidioides brasiliensis causes paracoccidioidomycosis, one of the most prevalent systemic mycosis in Latin America. Thus, understanding the characteristics of the protective immune response to P. brasiliensis is of interest, as it may reveal targets for disease control. The initiation of the immune response relies on the activation of pattern recognition receptors, among which are TLRs. Both TLR2 and TLR4 have been implicated in the recognition of P. brasiliensis and regulation of the immune response. However, the role of TLR9 during the infection by this fungus remains unclear. Methodology/Principal findings We used in vitro and in vivo models of infection by P. brasiliensis, comparing wild type and TLR9 deficient (−/−) mice, to assess the contribution of TLR9 on cytokine induction, phagocytosis and outcome of infection. We show that TLR9 recognizes either the yeast form or DNA from P. brasiliensis by stimulating the expression/production of pro-inflammatory cytokines by bone marrow derived macrophages, also increasing their phagocytic ability. We further show that TLR9 plays a protective role early after intravenous infection with P. brasiliensis, as infected TLR9−/− mice died at higher rate during the first 48 hours post infection than wild type mice. Moreover, TLR9−/− mice presented tissue damage and increased expression of several cytokines, such as TNF-α and IL-6. The increased pattern of cytokine expression was also observed during intraperitoneal infection of TLR9−/− mice, with enhanced recruitment of neutrophils. The phenotype of TLR9−/− hosts observed during the early stages of P. brasiliensis infection was reverted upon a transient, 48 hours post-infection, neutrophil depletion. Conclusions/Significance Our results suggest that TLR9 activation plays an early protective role against P. brasiliensis, by avoiding a deregulated type of inflammatory response associated to neutrophils that may lead to tissue damage. Thus

  18. Two-dimensional electrophoresis and characterization of antigens from Paracoccidioides brasiliensis.

    PubMed

    da Fonseca, C A; Jesuino, R S; Felipe, M S; Cunha, D A; Brito, W A; Soares, C M

    2001-06-01

    Paracoccidioides brasiliensis is a fungal pathogen of humans. To identify antigens from P. brasiliensis we fractionated a crude preparation of proteins from the fungus and detected the IgG reactive proteins by immunoblot assays of yeast cellular extracts with sera of patients with paracoccidioidomycosis (PCM). We identified and characterized six new antigens by amino acid sequencing and homology search analyses with other proteins deposited in a database. The newly characterized antigens were highly homologous to catalase, fructose-1,6-biphosphate aldolase (aldolase), glyceraldehyde-3-phosphate dehydrogenase, malate dehydrogenase and triosephosphate isomerase from several sources. The characterized antigens presented preferential synthesis in yeast cells, the host fungus phase. PMID:11418327

  19. Immunomodulating Activity of Agaricus brasiliensis KA21 in Mice and in Human Volunteers

    PubMed Central

    Fukuwatari, Yasushi; Okumura, Ko; Takeda, Kazuyoshi; Ishibashi, Ken-ichi; Furukawa, Mai; Ohno, Naohito; Mori, Kazu; Gao, Ming; Motoi, Masuro

    2008-01-01

    We performed studies on murine models and human volunteers to examine the immunoenhancing effects of the naturally outdoor-cultivated fruit body of Agaricus brasiliensis KA21 (i.e. Agaricus blazei). Antitumor, leukocyte-enhancing, hepatopathy-alleviating and endotoxin shock-alleviating effects were found in mice. In the human study, percentage body fat, percentage visceral fat, blood cholesterol level and blood glucose level were decreased, and natural killer cell activity was increased. Taken together, the results strongly suggest that the A. brasiliensis fruit body is useful as a health-promoting food. PMID:18604247

  20. Therapeutic Effect of Agaricus brasiliensis on Phenylhydrazine-Induced Neonatal Jaundice in Rats

    PubMed Central

    Zhang, Lan; Yuan, Bo; Wang, HuiPing; Gao, Ya

    2015-01-01

    The present study was designed to investigate the effect of Agaricus brasiliensis extract (ABE) on phenylhydrazine-induced neonatal jaundice in rats. Administration of ABE dose-dependently reduced the elevated bilirubin level induced by phenylhydrazine. It can be somewhat supported from the results of in vitro bilirubin degradation experiment. ABE treatment also reduced the total antioxidant status (TAOS), cascade O2−/SOD, level of NF-κB protein, and adrenomedullin (AM). Overall, the results of this study demonstrated that Agaricus brasiliensis extract may be beneficial to reducing bilirubin level without causing hepatotoxicity in neonatal jaundice. PMID:25883968

  1. Therapeutic effect of Agaricus brasiliensis on phenylhydrazine-induced neonatal jaundice in rats.

    PubMed

    Zhang, Lan; Yuan, Bo; Wang, HuiPing; Gao, Ya

    2015-01-01

    The present study was designed to investigate the effect of Agaricus brasiliensis extract (ABE) on phenylhydrazine-induced neonatal jaundice in rats. Administration of ABE dose-dependently reduced the elevated bilirubin level induced by phenylhydrazine. It can be somewhat supported from the results of in vitro bilirubin degradation experiment. ABE treatment also reduced the total antioxidant status (TAOS), cascade O2(-)/SOD, level of NF-κB protein, and adrenomedullin (AM). Overall, the results of this study demonstrated that Agaricus brasiliensis extract may be beneficial to reducing bilirubin level without causing hepatotoxicity in neonatal jaundice. PMID:25883968

  2. Production of Paracoccidioides brasiliensis exoantigens for immunodiffusion tests.

    PubMed Central

    De Camargo, Z; Unterkircher, C; Campoy, S P; Travassos, L R

    1988-01-01

    Growth curves of the yeast form of Paracoccidioides brasiliensis B-339 based on total and viable cell counts were determined. Crude culture filtrate antigens were obtained after 7, 10, 15, 20, 25, and 30 days of incubation. Different patterns of proteins were obtained by affinity chromatography on Sepharose 4B-immunoglobulin G complex made with immunoglobulin G from patients with paracoccidioidomycosis, with subsequent analyses by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and scanning densitometry. Three major proteins were excreted during the time course of a 30-day culture: a doublet at 20 to 21 kilodaltons (kDa) and molecules of 43 and 52 kDa. The 43-kDa antigen was present throughout the growth period, and its level reached a peak on days 15 to 20 and then decreased considerably toward day 30. The antigenic preparations collected on days 7, 10, 15, and 20 gave better reactions in immunodiffusion tests than those collected on days 25 and 30. The 7-day exoantigen gave a sensitivity of 97.1% and specificity of 100% on immunodiffusion. The main line of precipitation had a very high intensity, showing a total identity with that of a previously purified glycoprotein of 43 kDa. A 7-day crude exoantigen displayed a high level of sensitivity and specificity, being reproducible from batch to batch and retaining its activity for years when kept lyophilized. A protocol is recommended for the production of a stable diagnostic antigen to be used in immunodiffusion tests for paracoccidioidomycosis. Images PMID:3141460

  3. A Morphological and Cytochemical Study of the Interaction between Paracoccidiodes brasiliensis and Neutrophils

    NASA Astrophysics Data System (ADS)

    Dias, Maria Fernanda R. G.; Filgueira, Absalom L.; de Souza, Wanderley

    2004-04-01

    Paracoccidioidomycosis is a systemic granulomatous disease caused by the dimorphic fungus Paracoccidioides brasiliensis. It is the most prevalent systemic mycosis of Latin America and 80% of the reported cases are from Brazil. Because of the great number of neutrophils found in the P. brasiliensis granuloma, studies have been done to evaluate the role of these cells during the development of the infection. Scanning and transmission electron microscopy of thin sections showed that the neutrophils ingest yeast cells through a typical phagocytic process with the formation of pseudopodes. The pseudopodes even disrupt the connection established between the mother and the bud cells. Neutrophils also associate to each other, forming a kind of extracellular vacuole where large yeast cells are encapsulated. Cytochemical studies showed that once P. brasiliensis attaches to the neutrophil surface, it triggers a respiratory burst with release of oxygen-derived products. Attachment also triggers neutrophils' degranulation, with release of endogenous peroxidase localized in cytoplasmic granules. Together, these processes lead to killing of both ingested and extracellular P. brasiliensis.

  4. Initial development of the endocarp in Lithraea brasiliensis Marchand (Anacardiaceae): with taxonomic notes.

    PubMed

    De Oliveira, João M S; Mariath, Jorge E A

    2015-09-01

    Investigation into the initial developmental stages of a given structure is fundamental for precise characterization as well as for comparative analysis in relation to other taxa when homologies are established. For the Anacardiaceae family, investigations of the initial development of the pericarp or its basic histological sites, the epicarp, mesocarp and endocarp, are relevant since these regions are of taxonomic and phylogenetic importance. The initial stages of endocarp development in Lithraea brasiliensis were studied using light microscopy. In L. brasiliensis, the fruits are of the drupe type. The endocarp originates exclusively in the epidermis of the locular cavity and is composed of only three strata. The crystalliferous layer, typical in fruits of the Anacardiaceae family, originates in the carpelar mesophyll and runs adjacent to the outermost layer of the endocarp. The endocarp in Lithraea brasiliensis is of the Anacardium type. The results of the present study are important for the identification of the Lithraea species that occur in the state of Rio Grande do Sul. In L. molleoides, the crystalliferous layer was shown to originate from the locular epidermis and this developmental aspect enables it to be differentiated from L. brasiliensis and demonstrates that there is variability of endocarp development in Lithraea. PMID:26221992

  5. A Novel Anelloviridae Species Detected in Tadarida brasiliensis Bats: First Sequence of a Chiropteran Anellovirus

    PubMed Central

    Cibulski, Samuel Paulo; Teixeira, Thais Fumaco; de Sales Lima, Francisco Esmaile; do Santos, Helton Fernandes; Franco, Ana Claudia

    2014-01-01

    Using metagenomic approaches, we identified a novel Torque teno virus from Brazilian free-tailed bats (Tadarida brasiliensis) (TT-TbV). The TT-TbV genome and deduced protein sequences share extremely low identity with known anelloviruses. Due to a high degree of phylogenetic divergence, such putative virus could not be allocated into any Anelloviridae genera. PMID:25359906

  6. Thermal preferences and limits of Triatoma brasiliensis in its natural environment - Field observations while host searching

    PubMed Central

    Catalá, Silvia; Bezerra, Claudia Mendonça; Diotaiuti, Lileia

    2015-01-01

    The goal of this work was to explore the thermal relationship between foraging Triatoma brasiliensis and its natural habitat during the hottest season in the state of Ceará, Brazil. The thermal profiles were determined using infrared analysis. Although the daily temperature of rock surfaces varied in a wide range, T. brasiliensis selected to walk through areas with temperatures between 31.7-40.5ºC. The temperature of T. brasiliensis body surface ranged from 32.8-34.4ºC, being higher in legs than the abdomen. A strong relationship was found between the temperature of the insect and the temperature of rock crevices where they were hidden (r: 0.96, p < 0.05). The species was active at full sunlight being a clear example of how the light-dark rhythm may be altered, even under predation risk. Our results strongly suggest a thermal borderline for T. brasiliensis foraging activity near 40ºC. The simultaneous determination of insect body and rock temperatures here presented are the only obtained in natural habitats for this or other triatomines. PMID:26517659

  7. Decreased expression of 14-3-3 in Paracoccidioides brasiliensis confirms its involvement in fungal pathogenesis.

    PubMed

    Marcos, Caroline Maria; Silva, Julhiany de Fátima ds; Oliveira, Haroldo Cesar de; Assato, Patrícia Akemi; Singulani, Junya de Lacorte; Lopez, Angela Maria; Tamayo, Diana Patricia; Hernandez-Ruiz, Orville; McEwen, Juan G; Mendes-Giannini, Maria José Soares; Fusco-Almeida, Ana Marisa

    2016-01-01

    The interaction between the fungal pathogen Paracoccidioides brasiliensis and host cells is usually mediated by specific binding events between adhesins on the fungal surface and receptors on the host extracellular matrix or cell surface. One molecule implicated in the P. brasiliensis-host interaction is the 14-3-3 protein. The 14-3-3 protein belongs to a family of conserved regulatory molecules that are expressed in all eukaryotic cells and are involved in diverse cellular functions. Here, we investigated the relevance of the 14-3-3 protein to the virulence of P. brasiliensis. Using antisense RNA technology and Agrobacterium tumefaciens-mediated transformation, we generated a 14-3-3-silenced strain (expression reduced by ˜55%). This strain allowed us to investigate the interaction between 14-3-3 and the host and to correlate the functions of P. brasiliensis 14-3-3 with cellular features, such as morphological characteristics and virulence, that are important for pathogenesis. PMID:26646480

  8. Topical anti-inflammatory activity of Eugenia brasiliensis Lam. (Myrtaceae) leaves.

    PubMed

    Pietrovski, Evelise Fernandes; Magina, Michele Debiasi Alberton; Gomig, Franciane; Pietrovski, Caroline Fernandes; Micke, Gustavo Amadeu; Barcellos, Michele; Pizzolatti, Moacir Geraldo; Cabrini, Daniela Almeida; Brighente, Inês Maria Costa; Otuki, Michel Fleith

    2008-04-01

    Eugenia brasiliensis Lam., a plant from the south of Brazil, is used in the popular medicine for rheumatism treatment. This study reports that topical application of hydroalcoholic extract, fractions and isolated compounds from E. brasiliensis caused an inhibition of ear oedema in response to topical application of croton oil on the mouse ear. For oedema inhibition, the estimated ID50 values (dose reducing the inflammatory response by 50% relative to the control value) for hydroalcoholic extract and fractions (hexane, ethyl acetate and dichloromethane) were 0.17, 0.29, 0.13 and 0.14 mg/ear, respectively, with inhibition of 79+/-7%, 87+/-6%, 88+/-5% and 96+/-2%, respectively. Isolated phenolic compounds (quercetin, catechin and gallocatechin) were also effective in inhibiting the oedema (inhibition of 61+/-5%, 66+/-2% and 37+/-9%, respectively). Moreover, both extract and isolated compounds caused inhibition of polymorphonuclear cells influx (inhibition of 85+/-6%, 81+/-5%, 73+/-6% and 76+/-6%, respectively). The histological analysis of the ear tissue clearly confirmed that the extract and compounds of E. brasiliensis inhibited the influx of polymorphonuclear cells to mouse ear skin after application of croton oil. Furthermore, hydroalcoholic extract was also effective in inhibiting the arachidonic acid-mediated mouse ear oedema (ID50 value was 1.94 mg/ear and inhibition of 60+/-7%). Therefore, these results consistently support the notion that E. brasiliensis possesses topical anti-inflammatory activity. PMID:18380921

  9. Activation of the alternative complement pathway in canine normal serum by Paracoccidioides brasiliensis

    PubMed Central

    Bianchini, A.A.C.; Petroni, T.F.; Fedatto, P.F.; Bianchini, R.R.; Venancio, E.J.; Itano, E.N.; Ono, M.A.

    2009-01-01

    The dimorphic fungus Paracoccidioides brasiliensis is the etiological agent of paracoccidioidomycosis, a human granulomatous disease. Recently the first case of natural disease in dogs was reported. The complement system is an important effector component of humoral immunity against infectious agents. Therefore, the aim of this study was to evaluate the activation of the dog alternative complement pathway by P. brasiliensis. Initially, the ability of erythrocytes of guinea pig, rabbit, sheep, chicken and swine to activate the dog alternative pathway was evaluated. The guinea pig erythrocytes showed the greatest capacity to activate dog alternative pathway. The alternative (AH50) hemolytic activity was evaluated in 27 serum samples from healthy dogs and the mean values were 87.2 AH50/ml. No significant differences were observed in relation to sex and age. The alternative pathway activation by P. brasiliensis was higher in serum samples from adult dogs when compared to puppies and aged dogs (p ≤ 0.05). This is the first report of dog alternative complement pathway activation by P. brasiliensis and suggests that it may play a protective role in canine paracoccidioidomycosis. PMID:24031350

  10. National Plant Diagnostic Network, Taxonomic training videos: Aphids under the microscope - Cerataphis brasiliensis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Training is a critical part of aphid (Hemiptera: Aphididae) identification. This video provides provides training to identify the palm aphid, Cerataphis brasiliensis, using a compound microscope and an electronic identification key called “LUCID.” The video demonstrates key morphological structures...

  11. Do different degrees of human activity affect the diet of Brazilian silverside Atherinella brasiliensis?

    PubMed

    Alves, V E N; Patrício, J; Dolbeth, M; Pessanha, A; Palma, A R T; Dantas, E W; Vendel, A L

    2016-08-01

    The aim of the present study was to test whether different degrees of human activity affect the diet of the Brazilian silverside Atherinella brasiliensis in two tropical estuaries. Fish were collected along the salinity gradient of two Brazilian estuaries, the heavily impacted Paraiba Estuary and the less impacted Mamanguape Estuary, in the dry and wet seasons. The findings confirm that A. brasiliensis has generalist feeding habits and is able to change its diet under different environmental conditions. The results indicate clear spatial (i.e. along the estuarine gradient) changes in diet composition in both estuaries, but diet was also influenced by the degree of anthropogenic disturbance. During the wet season in the nutrient enriched Paraiba Estuary, when human activity was higher, the diet of A. brasiliensis was poorer and dominated by few dietary items, reflecting the potential impoverishment of prey items in this heavily disturbed system. The specimens collected in the most affected estuary also had a greater frequency of micro-plastics and parasites in their stomachs, reflecting the greater degree of human disturbance in the estuary. The present findings suggest that the diet of A. brasiliensis could be a useful indicator of changes in the ecological quality of these and other tropical estuaries of the western Atlantic Ocean. PMID:27328827

  12. Activation of the alternative complement pathway in canine normal serum by Paracoccidioides brasiliensis.

    PubMed

    Bianchini, A A C; Petroni, T F; Fedatto, P F; Bianchini, R R; Venancio, E J; Itano, E N; Ono, M A

    2009-04-01

    The dimorphic fungus Paracoccidioides brasiliensis is the etiological agent of paracoccidioidomycosis, a human granulomatous disease. Recently the first case of natural disease in dogs was reported. The complement system is an important effector component of humoral immunity against infectious agents. Therefore, the aim of this study was to evaluate the activation of the dog alternative complement pathway by P. brasiliensis. Initially, the ability of erythrocytes of guinea pig, rabbit, sheep, chicken and swine to activate the dog alternative pathway was evaluated. The guinea pig erythrocytes showed the greatest capacity to activate dog alternative pathway. The alternative (AH50) hemolytic activity was evaluated in 27 serum samples from healthy dogs and the mean values were 87.2 AH50/ml. No significant differences were observed in relation to sex and age. The alternative pathway activation by P. brasiliensis was higher in serum samples from adult dogs when compared to puppies and aged dogs (p ≤ 0.05). This is the first report of dog alternative complement pathway activation by P. brasiliensis and suggests that it may play a protective role in canine paracoccidioidomycosis. PMID:24031350

  13. IL-13 receptor alpha-2 regulates the immune and functional response to Nippostrongylus brasiliensis infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    IL-13 has a prominent role in host defense against the gastrointestinal nematode, Nippostrongylus brasiliensis; however, the role of IL-13 alpha2 in the immune and functional response to enteric infection is not known. In the current study, we investigated changes in smooth muscle and epithelial ce...

  14. Ecological study of Paracoccidioides brasiliensis in soil: growth ability, conidia production and molecular detection

    PubMed Central

    Terçarioli, Gisela Ramos; Bagagli, Eduardo; Reis, Gabriela Martins; Theodoro, Raquel Cordeiro; Bosco, Sandra De Moraes Gimenes; Macoris, Severino Assis da Graça; Richini-Pereira, Virgínia Bodelão

    2007-01-01

    Background Paracoccidioides brasiliensis ecology is not completely understood, although several pieces of evidence point to the soil as its most probable habitat. The present study aimed to investigate the fungal growth, conidia production and molecular pathogen detection in different soil conditions. Methods Soils samples of clayey, sandy and medium textures were collected from ground surface and the interior of armadillo burrows in a hyperendemic area of Paracoccidioidomycosis. P. brasiliensis was inoculated in soil with controlled humidity and in culture medium containing soil extracts. The molecular detection was carried out by Nested PCR, using panfungal and species specific primers from the ITS-5.8S rDNA region. Results The soil texture does not affect fungus development and the growth is more abundant on/in soil saturated with water. Some soil samples inhibited the development of P. brasiliensis, especially those that contain high values of Exchangeable Aluminum (H+Al) in their composition. Some isolates produced a large number of conidia, mainly in soil-extract agar medium. The molecular detection was positive only in samples collected from armadillo burrows, both in sandy and clayey soil. Conclusion P. brasiliensis may grow and produce the infectious conidia in sandy and clayey soil, containing high water content, mainly in wild animal burrows, but without high values of H+Al. PMID:17953742

  15. Epidemiology of Ornithodoros brasiliensis (mouro tick) in the southern Brazilian highlands and the description of human and animal retrospective cases of tick parasitism.

    PubMed

    Reck, José; Marks, Fernanda S; Guimarães, Jorge A; Termignoni, Carlos; Martins, João Ricardo

    2013-02-01

    Ornithodoros brasiliensis, also known as the "mouro" tick, is an argasid tick found exclusively in the southern Brazilian highlands. O. brasiliensis parasitism is frequently associated with severe symptoms directly induced by the tick bite, a condition compatible with the definition of tick toxicosis. The objectives of this work include (i) the determination of the distribution of O. brasiliensis in farms located in the tick-endemic region, (ii) the description of the characteristics of O. brasiliensis habitats, (iii) the analysis of risk factors associated with O. brasiliensis, and (iv) the retrospective description of cases of human and animal parasitism by O. brasiliensis. Of the 30 farms included in this study, O. brasiliensis was identified on 5 farms (frequency 16.7%), in which several ticks found in high density buried in soil were collected. Information regarding the tick habitats and the local population was recorded. The data indicated that O. brasiliensis feeds on humans, dogs, armadillos (Dasypus hybridus), and possibly skunks (Conepatus chinga). The analysis of risk factors indicated that the presence of house basements with an unpaved (natural soil) floor on farms and insufficient sanitary conditions significantly enhanced the probability of identifying O. brasiliensis. Additionally, we describe retrospectively cases of tick parasitism in 28 humans and 11 dogs including the most common symptoms associated with tick toxicosis. This is the first study concerning O. brasiliensis epidemiology, distribution, and habitat, and the report represents the most comprehensive characterization of Ornithodoros bite-associated toxicosis syndrome. PMID:23238249

  16. Study of nucleolar behavior during spermatogenesis in Martarega brasiliensis (Heteroptera, Notonectidae).

    PubMed

    Pereira, L L V; Alevi, K C C; Moreira, F F F; Barbosa, J F; Silistino-Souza, E R; Silva Júnior, F C; Souza-Firmino, T S; Banho, C A; Itoyama, M M

    2015-01-01

    Few cytogenetic studies have been undertaken using aquatic heteropterans and the nucleolar behavior of these insects has been described in only four species, Limnogonus aduncus, Brachymetra albinerva, Halobatopsis platensis, and Cylindrostethus palmaris. The nucleolus is a cellular structure related to biosynthetic activity and it exhibits a peculiar behavior in the heteropterans of the Triatominae subfamily; it persists during all stages of meiosis. Thus, this study aims to analyze spermatogenesis in Martarega brasiliensis, with an emphasis on nucleolar behavior. Twenty M. brasiliensis adult males were used and collected from the Municipal reservoir in the city of São José do Rio Preto, São Paulo, Brazil. The species were fixed in methanol:acetic acid (3:1), then dissected, and the testicles were extracted, torn apart, and impregnated with silver ions. During prophase, the nuclei of M. brasiliensis were composed of the nucleolus and nucleolar corpuscles, which varied in number from one to four, emphasizing that this insect has great synthetic activity during meiosis. The analysis of cells in metaphase I showed that M. brasiliensis presents a nucleolar organizing region in at least one autosome. Furthermore, the phenomenon of nucleolar persistence was not observed. All spermatids presented nucleolar markings that varied in number and position according to the stage of elongation. Moreover, it was also possible to highlight the presence of a vesicle in spermatids. Thus, this paper describes the nucleolar behavior of M. brasiliensis and highlights important characteristics during spermatogenesis, thus, increasing the knowledge about the biology of these aquatic heteropterans. PMID:26345830

  17. Characterization of Paracoccidioides brasiliensis by FT-IR spectroscopy and nanotechnology.

    PubMed

    Ferreira, Isabelle; Ferreira-Strixino, Juliana; Castilho, Maiara L; Campos, Claudia B L; Tellez, Claudio; Raniero, Leandro

    2016-01-01

    Paracoccidioides brasiliensis, the etiological agent of paracoccidioidomycosis, is a dimorphic fungus existing as mycelia in the environment (or at 25°C in vitro) and as yeast cells in the human host (or at 37°C in vitro). Because mycological examination of lesions in patients frequently is unable to show the presence of the fungus and serological tests can misdiagnose the disease with other mycosis, the development of new approach's for molecular identification of P. brasiliensis spurges is needed. This study describes the use of a gold nanoprobe of a known gene sequence of P. brasiliensis as a molecular tool to identify P. brasiliensis by regular polymerase chain reaction (PCR) associated with a colorimetric methods. This approach is suitable for testing in remote areas because it does not require any further step than gene amplification, being safer and cheaper than electrophoresis methods. The proposed test showed a color change of the PCR reaction mixture from red to blue in negative samples, whereas the solution remains red in positive samples. We also performed a Fourier Transform Infrared (FT-IR) Spectroscopy analysis to characterize and compare the chemical composition between yeast and mycelia forms, which revealed biochemical differences between these two forms. The analysis of the spectra showed that differences were distributed in chemical bonds of proteins, lipids and carbohydrates. The most prominent difference between both forms was vibration modes related to 1,3-β-glucan usually found in mycelia and 1,3-α-glucan found in yeasts and also chitin forms. In this work, we introduce FT-IR as a new method suitable to reveal overall differences that biochemically distinguish each form of P. brasiliensis that could be additionally used to discriminate biochemical differences among a single form under distinct environmental conditions. PMID:26241825

  18. Characterization of Paracoccidioides brasiliensis by FT-IR spectroscopy and nanotechnology

    NASA Astrophysics Data System (ADS)

    Ferreira, Isabelle; Ferreira-Strixino, Juliana; Castilho, Maiara L.; Campos, Claudia B. L.; Tellez, Claudio; Raniero, Leandro

    2016-01-01

    Paracoccidioides brasiliensis, the etiological agent of paracoccidioidomycosis, is a dimorphic fungus existing as mycelia in the environment (or at 25 °C in vitro) and as yeast cells in the human host (or at 37 °C in vitro). Because mycological examination of lesions in patients frequently is unable to show the presence of the fungus and serological tests can misdiagnose the disease with other mycosis, the development of new approach's for molecular identification of P. brasiliensis spurges is needed. This study describes the use of a gold nanoprobe of a known gene sequence of P. brasiliensis as a molecular tool to identify P. brasiliensis by regular polymerase chain reaction (PCR) associated with a colorimetric methods. This approach is suitable for testing in remote areas because it does not require any further step than gene amplification, being safer and cheaper than electrophoresis methods. The proposed test showed a color change of the PCR reaction mixture from red to blue in negative samples, whereas the solution remains red in positive samples. We also performed a Fourier Transform Infrared (FT-IR) Spectroscopy analysis to characterize and compare the chemical composition between yeast and mycelia forms, which revealed biochemical differences between these two forms. The analysis of the spectra showed that differences were distributed in chemical bonds of proteins, lipids and carbohydrates. The most prominent difference between both forms was vibration modes related to 1,3-β-glucan usually found in mycelia and 1,3-α-glucan found in yeasts and also chitin forms. In this work, we introduce FT-IR as a new method suitable to reveal overall differences that biochemically distinguish each form of P. brasiliensis that could be additionally used to discriminate biochemical differences among a single form under distinct environmental conditions.

  19. Cloning, killing, and identity.

    PubMed Central

    McMahan, J

    1999-01-01

    One potentially valuable use of cloning is to provide a source of tissues or organs for transplantation. The most important objection to this use of cloning is that a human clone would be the sort of entity that it would be seriously wrong to kill. I argue that entities of the sort that you and I essentially are do not begin to exist until around the seventh month of fetal gestation. Therefore to kill a clone prior to that would not be to kill someone like you or me but would be only to prevent one of us from existing. And even after one of us begins to exist, the objections to killing it remain comparatively weak until its psychological capacities reach a certain level of maturation. These claims support the permissibility of killing a clone during the early stages of its development in order to use its organs for transplantation. PMID:10226909

  20. Immunity-dependent reduction of segmented filamentous bacteria in mice infected with the helminthic parasite Nippostrongylus brasiliensis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Immune modulation by helminth (worm) parasites could protect the host against autoimmune diseases. We report that the parasitic nematode Nippostrongylus brasiliensis induces changes in the expression of antimicrobial peptides that are associated with marked microbial composition shifts, including re...

  1. Changes in kinematics and aerodynamics over a range of speeds in Tadarida brasiliensis, the Brazilian free-tailed bat

    PubMed Central

    Hubel, Tatjana Y.; Hristov, Nickolay I.; Swartz, Sharon M.; Breuer, Kenneth S.

    2012-01-01

    To date, wake measurements using particle image velocimetry (PIV) of bats in flight have studied only three bat species, all fruit and nectar feeders. In this study, we present the first wake structure analysis for an insectivorous bat. Tadarida brasiliensis, the Brazilian free-tailed bat, is an aerial hunter that annually migrates long distances and also differs strikingly from the previously investigated species morphologically. We compare the aerodynamics of T. brasiliensis with those of other, frugivorous bats and with common swifts, Apus apus, a bird with wing morphology, kinematics and flight ecology similar to that of these bats. The comparison reveals that, for the range of speeds evaluated, the cyclical pattern of aerodynamic forces associated with a wingbeat shows more similarities between T. brasiliensis and A. apus than between T. brasiliensis and other frugivorous bats. PMID:22258554

  2. Ethrel-stimulated prolongation of latex flow in the rubber tree (Hevea brasiliensis Muell. Arg.): an Hev b 7-like protein acts as a universal antagonist of rubber particle aggregating factors from lutoids and C-serum.

    PubMed

    Shi, Min-Jing; Cai, Fu-Ge; Tian, Wei-Min

    2016-02-01

    Ethrel is the most effective stimuli in prolonging the latex flow that consequently increases yield per tapping. This effect is largely ascribed to the enhanced lutoid stability, which is associated with the decreased release of initiators of rubber particle (RP) aggregation from lutoid bursting. However, the increase in both the bursting index of lutoids and the duration of latex flow after applying ethrel or ethylene gas in high concentrations suggests that a new mechanism needs to be introduced. In this study, a latex allergen Hev b 7-like protein in C-serum was identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF MS). In vitro analysis showed that the protein acted as a universal antagonist of RP aggregating factors from lutoids and C-serum. Ethrel treatment obviously weakened the effect of C-serum on RP aggregation, which was closely associated with the increase in the level of the Hev b 7-like protein and the decrease in the level of the 37 kDa protein, as revealed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), western blotting analysis and antibody neutralization. Thus, the increase of the Hev b 7-like protein level or the ratio of the Hev b 7-like protein to the 37 kDa protein in C-serum should be primarily ascribed to the ethrel-stimulated prolongation of latex flow duration. PMID:26381537

  3. Morphometric and high resolution scanning electron microscopy analysis of low-level laser therapy and latex protein (Hevea brasiliensis) administration following a crush injury of the sciatic nerve in rats.

    PubMed

    Dias, Fernando J; Issa, João Paulo M; Coutinho-Netto, Joaquim; Fazan, Valéria P S; Sousa, Luiz Gustavo; Iyomasa, Mamie M; Papa, Paula C; Watanabe, Ii-Sei

    2015-02-15

    This study evaluated the effect of low-level laser therapy (LLLT; 15 J/cm(2)) and a latex protein (F1) on a crush injury of the sciatic (ischiadicus) nerve. Seventy-two rats (male, 250 g) were divided into 6 groups: CG, control; EG, exposed nerve; IG, injured nerve without treatment; LG, injured nerve with LLLT; HG, injured nerve with F1; and LHG, injured nerve with LLLT and F1. After 4 or 8 weeks, the animals were euthanized and samples of the sciatic nerve were collected for morphometric and high-resolution scanning electron microscopy (HRSEM) analysis. After 4 weeks, the morphometry revealed improvements in the treated animals, and the HG appeared to be the most similar to the CG; after 8 weeks, the injured groups showed improvements compared to the previous period, and the results of the treatment groups were more similar to one another. At HRSEM after 4 weeks, the treated groups were similar and showed improvement compared to the IG; after 8 weeks, the LHG and HG had the best results. In conclusion, the treatments resulted in improvement after the nerve injury, and this recovery was time-dependent. In addition, the use of the F1 resulted in the best morphometric and ultrastructural findings. PMID:25619570

  4. Paracoccidioides brasiliensis Enolase Is a Surface Protein That Binds Plasminogen and Mediates Interaction of Yeast Forms with Host Cells ▿

    PubMed Central

    Nogueira, Sarah Veloso; Fonseca, Fernanda L.; Rodrigues, Marcio L.; Mundodi, Vasanth; Abi-Chacra, Erika A.; Winters, Michael S.; Alderete, John F.; Soares, Célia Maria de Almeida

    2010-01-01

    Paracoccidioidomycosis (PCM), caused by the dimorphic fungus Paracoccidioides brasiliensis, is a disseminated, systemic disorder that involves the lungs and other organs. The ability of the pathogen to interact with host components, including extracellular matrix (ECM) proteins, is essential to further colonization, invasion, and growth. Previously, enolase (EC 4.2.1.11) was characterized as a fibronectin binding protein in P. brasiliensis. Interaction of surface-bound enolase with plasminogen has been incriminated in tissue invasion for pathogenesis in several pathogens. In this paper, enolase was expressed in Escherichia coli as a recombinant glutathione S-transferase (GST) fusion protein (recombinant P. brasiliensis enolase [rPbEno]). The P. brasiliensis native enolase (PbEno) was detected at the fungus surface and cytoplasm by immunofluorescence with an anti-rPbEno antibody. Immobilized purified rPbEno bound plasminogen in a specific, concentration-dependent fashion. Both native enolase and rPbEno activated conversion of plasminogen to plasmin through tissue plasminogen activator. The association between PbEno and plasminogen was lysine dependent. In competition experiments, purified rPbEno, in its soluble form, inhibited plasminogen binding to fixed P. brasiliensis, suggesting that this interaction required surface-localized PbEno. Plasminogen-coated P. brasiliensis yeast cells were capable of degrading purified fibronectin, providing in vitro evidence for the generation of active plasmin on the fungus surface. Exposure of epithelial cells and phagocytes to enolase was associated with an increased expression of surface sites of adhesion. In fact, the association of P. brasiliensis with epithelial cells and phagocytes was increased in the presence of rPbEno. The expression of PbEno was upregulated in yeast cells derived from mouse-infected tissues. These data indicate that surface-associated PbEno may contribute to the pathogenesis of P. brasiliensis. PMID

  5. Human cord blood T-cell receptor alpha beta cell responses to protein antigens of Paracoccidioides brasiliensis yeast forms.

    PubMed Central

    Munk, M E; Kaufmann, S H

    1995-01-01

    Paracoccidioides brasiliensis causes a chronic granulomatous mycosis, prevalent in South America, and cell-mediated immunity represents the principal mode of protection against this fungal infection. We investigated the response of naive cord blood T cells to P. brasiliensis lysates. Our results show: (1) P. brasiliensis stimulates T-cell expansion, interleukin-2 (IL-2) production and differentiation into cytotoxic T cells; (2) T-cell stimulation depends on P. brasiliensis processing and major histocompatibility complex (MHC) class II expression; (3) the responsive T-cell population expresses alpha beta T-cell receptors (TCR) with different V beta gene products, CD4 and CD45RO; (4) the P. brasiliensis components involved in T-cell expansion primarily reside in a high molecular weight (100,000 MW) and a low molecular weight (< 1000 MW) protein fraction. These results indicate that protein antigens of P. brasiliensis stimulate cord blood CD4 alpha beta T cells, independent from in vivo presensitization, and thus question direct correlation of positive in vitro responses with protective immunity in vivo. PMID:7890308

  6. Macrophage Interaction with Paracoccidioides brasiliensis Yeast Cells Modulates Fungal Metabolism and Generates a Response to Oxidative Stress

    PubMed Central

    Parente-Rocha, Juliana Alves; Parente, Ana Flávia Alves; Baeza, Lilian Cristiane; Bonfim, Sheyla Maria Rondon Caixeta; Hernandez, Orville; McEwen, Juan G.; Bailão, Alexandre Melo; Taborda, Carlos Pelleschi; Borges, Clayton Luiz; Soares, Célia Maria de Almeida

    2015-01-01

    Macrophages are key players during Paracoccidioides brasiliensis infection. However, the relative contribution of the fungal response to counteracting macrophage activity remains poorly understood. In this work, we evaluated the P. brasiliensis proteomic response to macrophage internalization. A total of 308 differentially expressed proteins were detected in P. brasiliensis during infection. The positively regulated proteins included those involved in alternative carbon metabolism, such as enzymes involved in gluconeogenesis, beta-oxidation of fatty acids and amino acids catabolism. The down-regulated proteins during P. brasiliensis internalization in macrophages included those related to glycolysis and protein synthesis. Proteins involved in the oxidative stress response in P. brasiliensis yeast cells were also up-regulated during macrophage infection, including superoxide dismutases (SOD), thioredoxins (THX) and cytochrome c peroxidase (CCP). Antisense knockdown mutants evaluated the importance of CCP during macrophage infection. The results suggested that CCP is involved in a complex system of protection against oxidative stress and that gene silencing of this component of the antioxidant system diminished the survival of P. brasiliensis in macrophages and in a murine model of infection. PMID:26360774

  7. Interspecific interactions involving Neoseiulus californicus (Acari: Phytoseiidae) and Agistemus brasiliensis (Acari: Stigmaeidae) as predators of Brevipalpus phoenicis (Acari: Tenuipalpidae).

    PubMed

    da Silva, Marcos Zatti; Sato, Mário Eidi; de Oliveira, Carlos Amadeu Leite; Nicastro, Roberto Lomba

    2015-03-01

    Brevipalpus phoenicis (Geijskes) is associated with the transmission of Citrus leprosis which is considered the main viral disease for the Brazilian citrus production. Mites of the families Stigmaeidae and Phytoseiidae coexist in various agricultural crops, often promoting the biological control of pest mites. The aim of this work was to study the interactions of Neoseiulus californicus (McGregor) (Phytoseiidae) and Agistemus brasiliensis Matioli, Ueckermann & Oliveira (Stigmaeidae), in the presence or absence of B. phoenicis. Two experiments were carried out. In the first, a N. californicus female was placed in each leaf disc arena, with eggs of B. phoenicis and A. brasiliensis as food sources. In the second, an A. brasiliensis female was placed in each arena, with eggs of B. phoenicis and N. californicus as food sources. Adults of both predators were able to consume both types of eggs available as food sources, but they fed on considerably higher proportions of B. phoenicis than on eggs of the predator. Eggs of A. brasiliensis were not a suitable food source for N. californicus, which produced only 0.1 egg per female per day when only eggs of that species were present in the experimental unit. The results suggest that eggs of N. californicus were a suitable food source for A. brasiliensis, which oviposited 1.12 eggs per day, when only eggs of N. californicus were provided to the stigmaeid mite. The possible interactions among N. californicus, A. brasiliensis and B. phoenicis in citrus orchards are discussed. PMID:25524512

  8. Isolation of Paracoccidioides brasiliensis from armadillos (Dasypus novemcinctus) in an area where the fungus was recently isolated from soil.

    PubMed

    Silva-Vergara, M L; Martinez, R; Camargo, Z P; Malta, M H; Maffei, C M; Chadu, J B

    2000-06-01

    Natural infection of armadillos (Dasypus novemcinctus) with Paracoccidioides brasiliensis in Northern Brazil was reported in 1986, raising great interest in the understanding of the role of this mammal in the epidemiological cycle of the fungus. Recently, P. brasiliensis was isolated from the soil of Ibiá, State of Minas Gerais, southeastern Brazil. Armadillos captured in this area were evaluated for the presence of P. brasiliensis in the viscera and infection was detected in 4/16 animals (25%). Fungal yeast phase cells were observed in three of the four infected armadillos by direct microscopic examination and by the indirect immunofluorescence test carried out on homogenized tissues. P. brasiliensis was isolated from three armadillos whose homogenized viscera had been injected into Swiss mice. The new strains (Ibiá-T1, Ibiá-T2 and Ibiá-T3) were identified as P. brasiliensis on the basis of macro- and micromorphology, thermodimorphism, production and serologic activity of exoantigens, and by polymerase chain reaction (PCR)-detection of the gp43 gene. The lethality and lesions caused to the mice from which the strains were recovered confirmed the virulence of the isolates. We conclude that P. brasiliensis infects armadillos in locations with different geoclimatic characteristics and vegetation cover. The direct observation of yeast cells in tissues and the multiple visceral involvement, including the lungs, suggests the occurrence of paracoccidioidomycosis disease in these mammals and supports their role as wild hosts in the epidemiological cycle of the fungus. PMID:10892986

  9. Statement on Human Cloning

    MedlinePlus

    ... form Search American Association for the Advancement of Science Statement on Human Cloning Print Email Tweet The American Association for the Advancement of Science (AAAS) recognizes the intense debates within our society ...

  10. Do Managers Clone Themselves?

    ERIC Educational Resources Information Center

    Baron, Alma S.

    1981-01-01

    A recent questionnaire survey provides statistics on male managers' views of female managers. The author recommends that male managers break out of their cloning behavior and that the goal ought to be a plurality in management. (Author/WD)

  11. Disseminated Sporothrix brasiliensis Infection with Endocardial and Ocular Involvement in an HIV-Infected Patient

    PubMed Central

    Silva-Vergara, Mario León; de Camargo, Zoilo Pires; Silva, Patricia Ferreira; Abdalla, Michel Reis; Sgarbieri, Ricardo Nilsson; Rodrigues, Anderson Messias; dos Santos, Keila Cristina; Barata, Cristina Hueb; Ferreira-Paim, Kennio

    2012-01-01

    Disseminated sporotrichosis occurs in individuals with impaired cellular immunity, such as in cases of neoplasia, transplantation, diabetes, and especially, acquired immunodeficiency syndrome. This report presents a 32-year-old Brazilian human immunodeficiency virus (HIV)-infected patient who developed a protracted condition of disseminated sporotrichosis with endocarditis, bilateral endophthalmitis, and lymphatic involvement. He needed cardiac surgery to replace the mitral valve. Sporothrix brasiliensis isolates were recovered from cultures of subcutaneous nodules and mitral valve fragments. Species identification was based on classical and molecular methods. The patient received amphotericin B for 52 days and subsequently, oral itraconazole. He remains asymptomatic, and he is on maintenance therapy with itraconazole. Despite his positive clinical outcome, he developed bilateral blindness. To our knowledge, this case is the first report of endocarditis and endophthalmitis caused by S. brasiliensis. PMID:22403321

  12. Microsatellites for Carpotroche brasiliensis (Flacourtiaceae), a useful species for agroforestry and ecosystem conservation1

    PubMed Central

    Bittencourt, Flora; Alves, Jackeline S.; Gaiotto, Fernanda A.

    2015-01-01

    Premise of the study: We developed microsatellite markers for Carpotroche brasiliensis (Flacourtiaceae), a dioecious tree that is used as a food resource by midsize animals of the Brazilian fauna. Methods and Results: We designed 30 primer pairs using next-generation sequencing and classified 25 pairs as polymorphic. Observed heterozygosity ranged from 0.5 to 1.0, and expected heterozygosity ranged from 0.418 to 0.907. The combined probability of exclusion was greater than 0.999 and the combined probability of identity was less than 0.001, indicating that these microsatellites are appropriate for investigations of genetic structure, individual identification, and paternity testing. Conclusions: The developed molecular tools may contribute to future studies of population genetics, answering ecological and evolutionary questions regarding efficient conservation strategies for C. brasiliensis. PMID:26697275

  13. THE POWER OF THE SMALL: THE EXAMPLE OF Paracoccidioides brasiliensis CONIDIA

    PubMed Central

    RESTREPO, Angela; CANO, Luz Elena; GONZALEZ, Ángel

    2015-01-01

    SUMMARY Research on Paracoccidioides brasiliensis has centered in the yeast cell probably because of the lack of distinctive features in the mycelium. In 1942 and for the first time, lateral conidia were noticed in the fungus' hyphae. Later on, Brazilian, Venezuelan and Argentinean researchers described "aleurias" when the fungus was grown in natural substrates. In 1970 authors became interested in the conidia and were able to obtain them in large numbers and treat them as individual units. Their shape and size were defined and the presence of all the elements of a competent eukaryotic cell were demonstrated. Conidia exhibited thermal dimorphism and, additionally, when given intranasally to BALB/c male mice, they converted into yeasts in the lungs and produce progressive pulmonary lesions with further dissemination to other organs. Studies on the phagocyte-conidia interaction were revealing and showed that these versatile structures allow a better understanding of the host- P. brasiliensis interactions. PMID:26465363

  14. Mutants of Paracoccidioides brasiliensis strain IVIC Pb9 affected in dimorphism.

    PubMed

    San-Blas, F; San-Blas, G

    1992-01-01

    Morphological mutants were isolated after nitrosoguanidine treatment of Paracoccidioides brasiliensis strain IVIC Pb9. Two of these mutants, Pb257 and Pb258, developed a typical mycelia at 23 degrees C, however, the yeast cells which developed at 37 degrees C were indistinguishable from those of the parental strain. A third mutant, strain Pb267, was thermosensitive, grew as yeast-like cells at 23 degrees C, but was unable to survive at 37 degrees C. Morphological observations as well as serological and segregation tests confirmed that the mutant strains originated from P. brasiliensis. Cell wall chemical analyses of the mutant strains grown at 23 degrees C indicated the presence of alkali-soluble, acid-insoluble polysaccharides absent in the parental wild-type strain Pb9 grown under the same conditions. The phenotypes shown by the mutant strains may be related to deficiencies in the proper synthesis of cell wall components of the mycelial phase of this fungus. PMID:1573521

  15. Performance and meat quality of broiler chickens that are fed diets supplemented with Agaricus brasiliensis mushrooms.

    PubMed

    Guimarães, João Borges; Dos Santos, Eder Clementino; Dias, Eustáquio Souza; Bertechini, Antônio Gilberto; da Silva Ávila, Carla Luiza; Dias, Francesca Silva

    2014-12-01

    This trial was performed to study the use of the mushroom Agaricus brasiliensis as an alternative additive to antimicrobial growth promoters in broiler chicken diets and to assess the quality of the broiler chicken breast meat of birds that are fed diets containing this fungus. Thus, 595 1-day-old chicks were reared in reused poultry litter without anticoccidial and antimicrobial additives. The results showed that a concentration of 1.6 g mushrooms/kg diet was ideal for these birds because it provided better bird performance. When the birds' immune system organs were analyzed, it was found that the addition of both mushrooms influenced the immune system organs of these broiler chickens. Adding A. brasiliensis to broiler chicken diets did not compromise breast meat quality. PMID:25169695

  16. Antifungal activity of fractions and two pure compounds of flowers from Wedelia paludosa (Acmela brasiliensis) (Asteraceae).

    PubMed

    Sartori, M R K; Pretto, J B; Cruz, A B; Bresciani, L F V; Yunes, R A; Sortino, M; Zacchino, S A; Cechinel, V Filho

    2003-08-01

    Wedelia paludosa (Acmela brasiliensis) (Asteraceae), a traditionally used native Brazilian medicinal plant, showed antifungal activity against dermatophytes in dilution tests. The hexane, dichloromethane and butanol fractions displayed activity against Epidermophyton floccosum, Trichophyton rubrum and Trichophyton mentagrophytes, with minimal inhibitory concentrations between 250 and 1000 microg/mL. Two pure compounds, identified as kaurenoic acid (1) and luteolin (2), also showed activity against these dermatophytes. PMID:12967035

  17. Isolation and antigenicity of a 45-kilodalton Paracoccidioides brasiliensis immunodominant antigen.

    PubMed Central

    Ferreira-da-Cruz, M F; Galvão-Castro, B; Daniel-Ribeiro, C

    1992-01-01

    In the present study, we analyzed human antibody responses to Paracoccidioides brasiliensis cellular antigens by the immunoblot technique to identify specific cellular components and to investigate the existence of antigen profile differences among serological responses of paracoccidioidomycosis (PCM) patients. Among the 64 PCM serum samples analyzed, a relatively homogeneous immunoglobulin G response to P. brasiliensis antigens was observed. The polypeptide with a mass of 45 kDa was the most clinically important, since antibody to this antigen was detectable in 90.6% of PCM patients studied and the six individuals who did not produce antibody were either at the end of treatment or in the posttherapy period and had shown clinical recovery. These facts suggested that the presence of this antibody may be an indicator of active disease. The 45-kDa antigen was also the most specific antigen of the PCM humoral immune response, since it reacted with only 2 of 79 (2.5%) heterologous serum samples tested: 1 histoplasmosis case and 1 tuberculosis case. This polypeptide was isolated from gels by electroelution and, when tested by an immunoradiometric assay and immunoblotting, maintained its reactivity with PCM sera and also with anti-P. brasiliensis polyclonal antibodies raised in rabbits at the same sensitivity levels as those obtained in immunoblotting with a crude antigen. Since in our assays the 45-kDa polypeptide was the major P. brasiliensis antigen and seemed to be specific for PCM, its use in alternative diagnostic methods is promising, especially in patients suspected of having the juvenile clinical form of PCM often associated with negative double-immunodiffusion results. Images PMID:1612736

  18. P. brasiliensis Virulence Is Affected by SconC, the Negative Regulator of Inorganic Sulfur Assimilation

    PubMed Central

    Menino, João Filipe; Saraiva, Margarida; Gomes-Rezende, Jéssica; Sturme, Mark; Pedrosa, Jorge; Castro, António Gil; Ludovico, Paula; Goldman, Gustavo H.; Rodrigues, Fernando

    2013-01-01

    Conidia/mycelium-to-yeast transition of Paracoccidioidesbrasiliensis is a critical step for the establishment of paracoccidioidomycosis, a systemic mycosis endemic in Latin America. Thus, knowledge of the factors that mediate this transition is of major importance for the design of intervention strategies. So far, the only known pre-requisites for the accomplishment of the morphological transition are the temperature shift to 37°C and the availability of organic sulfur compounds. In this study, we investigated the auxotrophic nature to organic sulfur of the yeast phase of Paracoccidioides, with special attention to P. brasiliensis species. For this, we addressed the role of SconCp, the negative regulator of the inorganic sulfur assimilation pathway, in the dimorphism and virulence of this pathogen. We show that down-regulation of SCONC allows initial steps of mycelium-to-yeast transition in the absence of organic sulfur compounds, contrarily to the wild-type fungus that cannot undergo mycelium-to-yeast transition under such conditions. However, SCONC down-regulated transformants were unable to sustain yeast growth using inorganic sulfur compounds only. Moreover, pulses with inorganic sulfur in SCONC down-regulated transformants triggered an increase of the inorganic sulfur metabolism, which culminated in a drastic reduction of the ATP and NADPH cellular levels and in higher oxidative stress. Importantly, the down-regulation of SCONC resulted in a decreased virulence of P. brasiliensis, as validated in an in vivo model of infection. Overall, our findings shed light on the inability of P. brasiliensis yeast to rely on inorganic sulfur compounds, correlating its metabolism with cellular energy and redox imbalances. Furthermore, the data herein presented reveal SconCp as a novel virulence determinant of P. brasiliensis. PMID:24066151

  19. Immunological Basis for the Gender Differences in Murine Paracoccidioides brasiliensis Infection

    PubMed Central

    Pinzan, Camila Figueiredo; Ruas, Luciana Pereira; Casabona-Fortunato, Anália Sulamita; Carvalho, Fernanda Caroline; Roque-Barreira, Maria-Cristina

    2010-01-01

    This study aimed to investigate the immunological mechanisms involved in the gender distinct incidence of paracoccidioidomycosis (pcm), an endemic systemic mycosis in Latin America, which is at least 10 times more frequent in men than in women. Then, we compared the immune response of male and female mice to Paracoccidioides brasiliensis infection, as well as the influence in the gender differences exerted by paracoccin, a P. brasiliensis component with carbohydrate recognition property. High production of Th1 cytokines and T-bet expression have been detected in the paracoccin stimulated cultures of spleen cells from infected female mice. In contrast, in similar experimental conditions, cells from infected males produced higher levels of the Th2 cytokines and expressed GATA-3. Macrophages from male and female mice when stimulated with paracoccin displayed similar phagocytic capability, while fungicidal activity was two times more efficiently performed by macrophages from female mice, a fact that was associated with 50% higher levels of nitric oxide production. In order to evaluate the role of sexual hormones in the observed gender distinction, we have utilized mice that have been submitted to gonadectomy followed by inverse hormonal reconstitution. Spleen cells derived from castrated males reconstituted with estradiol have produced higher levels of IFN-γ (1291±15 pg/mL) and lower levels of IL-10 (494±38 pg/mL), than normal male in response to paracoccin stimulus. In contrast, spleen cells from castrated female mice that had been treated with testosterone produced more IL-10 (1284±36 pg/mL) and less IFN-γ (587±14 pg/mL) than cells from normal female. In conclusion, our results reveal that the sexual hormones had a profound effect on the biology of immune cells, and estradiol favours protective responses to P. brasiliensis infection. In addition, fungal components, such as paracoccin, may provide additional support to the gender dimorphic immunity that marks P

  20. Morphological studies on the infraciliature of a planktonic ciliate, Tintinnopsis brasiliensis (Ciliophora: Tintinina)

    NASA Astrophysics Data System (ADS)

    Cai, Shengfang; Song, Weibo; Xu, Dapeng; Chiang, Kuoping

    2006-01-01

    A poorly-described marine planktonic ciliate, Tintinnopsis brasiliensis Kofoid & Campbell, 1929, collected from the Taiping Cape of Qingdao, China, was morphologically investigated based on permanent preparation after protargol impregnation and was compared with other related congeners. According to the infraciliature, three ciliary groups can be recognized, which reveals a very stable structure among specimens and denotes that the pattern of infraciliature is, apart from the features of the lorica, a highly reliable criterion for species identification.

  1. The sialotranscriptome of the blood-sucking bug Triatoma brasiliensis (Hemiptera, Triatominae)

    PubMed Central

    Santos, Adriana; Ribeiro, José Marcos C.; Lehane, Michael J.; Gontijo, Nelder Figueiredo; Veloso, Artur Botelho; Sant'Anna, Mauricio R.V.; Araujo, Ricardo Nascimento; Grisard, Edmundo C.; Pereira, Marcos Horácio

    2007-01-01

    Triatoma brasiliensis is the most important autochthon vector of Trypanosoma cruzi in Brazil, where it is widely distributed in the semiarid areas of the Northeast. In order to advance the knowledge of the salivary biomolecules of Triatominae, a salivary gland cDNA library of T. brasiliensis was mass sequenced and analyzed. Polypeptides were sequenced by HPLC/Edman degradation experiments. 1,712 cDNA sequences were obtained and grouped in 786 clusters. The housekeeping category had 24.4% and 17.8% of the clusters and sequences, respectively. The putatively secreted category contained 47.1% of the clusters and 68.2% of the sequences. Finally, 28.5% of the clusters, containing 14% of all sequences, were classified as unknown. The sialoma of T. brasiliensis showed a high amount and great variety of different lipocalins (93.8% of secreted proteins). Remarkably, a great number of serine proteases that were not observed in previous blood-sucking sialotranscriptomes were found. Nine Kazal peptides were identified, among them one with high homology to the tabanid vasodilator vasotab, suggesting that the Triatoma vasodilator could be a Kazal protein. PMID:17550826

  2. Insights into the plant polysaccharide degradation potential of the xylanolytic yeast Pseudozyma brasiliensis.

    PubMed

    Kaupert Neto, Antonio Adalberto; Borin, Gustavo Pagotto; Goldman, Gustavo Henrique; Damásio, André Ricardo de Lima; Oliveira, Juliana Velasco de Castro

    2016-03-01

    In second-generation (2G) bioethanol production, plant cell-wall polysaccharides are broken down to release fermentable sugars. The enzymes of this process are classified as carbohydrate-active enzymes (CAZymes) and contribute substantially to the cost of biofuel production. A novel basidiomycete yeast species, Pseudozyma brasiliensis, was recently discovered. It produces an endo-β-1,4-xylanase with a higher specific activity than other xylanases. This enzyme is essential for the hydrolysis of biomass-derived xylan and has an important role in 2G bioethanol production. In spite of the P. brasiliensis biotechnological potential, there is no information about how it breaks down polysaccharides. For the first time, we characterized the secretome of P. brasiliensis grown on different carbon sources (xylose, xylan, cellobiose and glucose) and also under starvation conditions. The growth and consumption of each carbohydrate and the activity of the CAZymes of culture supernatants were analyzed. The CAZymes found in its secretomes, validated by enzymatic assays, have the potential to hydrolyze xylan, mannan, cellobiose and other polysaccharides. The data show that this yeast is a potential source of hydrolases, which can be used for biomass saccharification. PMID:26712719

  3. Antioxidant Effects of Medicinal Mushrooms Agaricus brasiliensis and Ganoderma lucidum (Higher Basidiomycetes): Evidence from Animal Studies.

    PubMed

    Yurkiv, Borys; Wasser, Solomon P; Nevo, Eviatar; Sybirna, Nataliya O

    2015-01-01

    With diabetes mellitus and increased glucose concentrations, the mitochondria electron transport chain is disrupted, superoxide anions are overproduced, and oxidative stress develops in cells. Thus, preventing oxidative stress can produce a decrease in the antioxidant system activity and an increase in apoptosis in immune cells. The application of medicinal mushrooms is a new possible approach to diabetes mellitus treatment. Therefore, the aim of this work was to investigate the influence of administration of the medicinal mushrooms Agaricus brasiliensis and Ganoderma lucidum on antioxidant enzyme activity in rat leukocytes. Wistar outbred white rats were used in the study. Streptozotocin was intraperitoneally injected once at a dose of 50 mg/kg body weight. Mushroom preparations were orally administered at a dose of 1 g/kg/day for 2 weeks. This revealed that in diabetes mellitus, the level of antioxidant enzyme activity is significantly decreased compared with control values, whereas the levels of lipid peroxidation is increased; this manifested in an increase in the amount of thiobarbituric acid reactive substances (TBARS). The medicinal mushrooms' administration is accompanied by an increase in antioxidant enzyme activity to control values and is even higher in the case of A. brasiliensis administration when compared with the diabetic group. As for the indicators of lipid peroxidation under mushroom administration of A. brasiliensis and G. lucidum, we observed a significant decrease of TBARS levels compared with the diabetic group. Increased activity of antioxidant enzymes and reduction of TBARS level indicate pronounced antioxidant properties of studied mushrooms. PMID:26756186

  4. Morphometry, Bite-Force, and Paleobiology of the Late Miocene Caiman Purussaurus brasiliensis

    PubMed Central

    Aureliano, Tito; Ghilardi, Aline M.; Guilherme, Edson; Souza-Filho, Jonas P.; Cavalcanti, Mauro; Riff, Douglas

    2015-01-01

    Purussaurus brasiliensis thrived in the northwestern portion of South America during the Late Miocene. Although substantial material has been recovered since its early discovery, this fossil crocodilian can still be considered as very poorly understood. In the present work, we used regression equations based on modern crocodilians to present novel details about the morphometry, bite-force and paleobiology of this species. According to our results, an adult Purussaurus brasiliensis was estimated to reach around 12.5 m in length, weighing around 8.4 metric tons, with a mean daily food intake of 40.6 kg. It was capable of generating sustained bite forces of 69,000 N (around 7 metric tons-force). The extreme size and strength reached by this animal seems to have allowed it to include a wide range of prey in its diet, making it a top predator in its ecosystem. As an adult, it would have preyed upon large to very large vertebrates, and, being unmatched by any other carnivore, it avoided competition. The evolution of a large body size granted P. brasiliensis many advantages, but it may also have led to its vulnerability. The constantly changing environment on a large geological scale may have reduced its long-term survival, favoring smaller species more resilient to ecological shifts. PMID:25689140

  5. Morphometry, bite-force, and paleobiology of the late miocene caiman Purussaurus brasiliensis.

    PubMed

    Aureliano, Tito; Ghilardi, Aline M; Guilherme, Edson; Souza-Filho, Jonas P; Cavalcanti, Mauro; Riff, Douglas

    2015-01-01

    Purussaurus brasiliensis thrived in the northwestern portion of South America during the Late Miocene. Although substantial material has been recovered since its early discovery, this fossil crocodilian can still be considered as very poorly understood. In the present work, we used regression equations based on modern crocodilians to present novel details about the morphometry, bite-force and paleobiology of this species. According to our results, an adult Purussaurus brasiliensis was estimated to reach around 12.5 m in length, weighing around 8.4 metric tons, with a mean daily food intake of 40.6 kg. It was capable of generating sustained bite forces of 69,000 N (around 7 metric tons-force). The extreme size and strength reached by this animal seems to have allowed it to include a wide range of prey in its diet, making it a top predator in its ecosystem. As an adult, it would have preyed upon large to very large vertebrates, and, being unmatched by any other carnivore, it avoided competition. The evolution of a large body size granted P. brasiliensis many advantages, but it may also have led to its vulnerability. The constantly changing environment on a large geological scale may have reduced its long-term survival, favoring smaller species more resilient to ecological shifts. PMID:25689140

  6. Participation of dectin-1 receptor on NETs release against Paracoccidioides brasiliensis: Role on extracellular killing.

    PubMed

    Bachiega, Tatiana Fernanda; Dias-Melicio, Luciane Alarcão; Fernandes, Reginaldo Keller; de Almeida Balderramas, Helanderson; Rodrigues, Daniela Ramos; Ximenes, Valdecir Farias; de Campos Soares, Ângela Maria Victoriano

    2016-02-01

    Paracoccidioides brasiliensis is a dimorphic fungus from the Paracoccidioides genus, which is the causative agent of paracoccidioidomycosis, a chronic, subacute or acute mycosis, with visceral and cutaneous involvement. This disease that is acquired through inhalation primarily attacks the lungs but, can spread to other organs. Phagocytic cells as neutrophils play an important role during innate immune response against this fungus, but studies on antifungal activities of these cells are scarce. In addition to their ability to eliminate pathogens by phagocytosis and antimicrobial secretions, neutrophils can trap and kill microorganisms by release of extracellular structures composed by DNA and antimicrobial proteins, called neutrophil extracellular traps (NETs). Here, we provide evidence that P. brasiliensis virulent strain (P. brasiliensis 18) induces NETs release. These structures were well evidenced by scanning electron microscopy, and specific NETs compounds such as histone, elastase and DNA were shown by confocal microscopy. In addition, we have shown that dectin-1 receptor is the main PRR to which fungus binds to induce NETS release. Fungi were ensnared by NETs, denoting the role of these structures in confining the fungus, avoiding dissemination. NETs were also shown to be involved in fungus killing, since fungicidal activity detected before and mainly after neutrophils activation with TNF-α, IFN-γ and GM-CSF was significantly inhibited by cocultures treatment with DNAse. PMID:26416210

  7. Lack of Galectin-3 Drives Response to Paracoccidioides brasiliensis toward a Th2-Biased Immunity

    PubMed Central

    Ruas, Luciana Pereira; Bernardes, Emerson Soares; Fermino, Marise Lopes; de Oliveira, Leandro Licursi; Hsu, Daniel K.; Liu, Fu-Tong; Chammas, Roger; Roque-Barreira, Maria-Cristina

    2009-01-01

    There is recent evidence that galectin-3 participates in immunity to infections, mostly by tuning cytokine production. We studied the balance of Th1/Th2 responses to P. brasiliensis experimental infection in the absence of galectin-3. The intermediate resistance to the fungal infection presented by C57BL/6 mice, associated with the development of a mixed type of immunity, was replaced with susceptibility to infection and a Th2-polarized immune response, in galectin-3-deficient (gal3−/−) mice. Such a response was associated with defective inflammatory and delayed type hypersensitivity (DTH) reactions, high IL-4 and GATA-3 expression and low nitric oxide production in the organs of infected animals. Gal3−/− macrophages exhibited higher TLR2 transcript levels and IL-10 production compared to wild-type macrophages after stimulation with P. brasiliensis antigens. We hypothesize that, during an in vivo P. brasiliensis infection, galectin-3 exerts its tuning role on immunity by interfering with the generation of regulatory macrophages, thus hindering the consequent Th2-polarized type of response. PMID:19229338

  8. Response of Paracoccidioides brasiliensis Pb01 to stressor agents and cell wall osmoregulators.

    PubMed

    Tomazett, Patrícia Kott; Castro, Nadya da Silva; Lenzi, Henrique Leonel; de Almeida Soares, Célia Maria; Pereira, Maristela

    2011-01-01

    In its attempt to survive, the fungal cell can change the cell wall composition and/or structure in response to environmental stress. The molecules involved in these compensatory mechanisms are a possible target for the development of effective antifungal agents. In the thermodimorphic fungus Paracoccidioides brasiliensis Pb01, the main polymers that compose the cell wall are chitin and glucans. These polymers form a primary barrier that is responsible for the structural integrity and formation of the cell wall. In this study the behaviour of P. brasiliensis was evaluated under incubation with cell wall stressor agents such as Calcofluor White (CFW), Congo Red (CR), Sodium Dodecyl Sulphate (SDS), NaCl, KCl, and Sorbitol. Use of concentrations at which the fungus is visually sensitive to those agents helped to explain some of the adaptive mechanisms used by P. brasiliensis in response to cell wall stress. Our results show that 1,3-β-D-glucan synthase (PbFKS1), glucosamine-6-phosphate synthase (PbGFA1) and β-1,3-glucanosyltransferase (PbGEL3)as well as 1,3-β-D-glucan and N-acetylglucosamine (GlcNAc) residues in the cell wall are involved in compensatory mechanisms against cell wall damage. PMID:21215956

  9. Control of Linepithema micans (Hymenoptera: Formicidae) and Eurhizococcus brasiliensis (Hemiptera: Margarodidae) in Vineyards Using Toxic Baits.

    PubMed

    Nondillo, Aline; Andzeiewski, Simone; Bello Fialho, Flávio; Bueno, Odair Correa; Botton, Marcos

    2016-08-01

    Linepithema micans (Forel) (Hymenoptera: Formicidae) is the main ant species responsible for dispersal of Eurhizococcus brasiliensis (Wille) (Hemiptera: Margarodidae), a root scale that damages grapevines in southern Brazil. The effects of different formulations of toxic baits based on boric acid and hydramethylnon to control L. micans and E. brasiliensis were evaluated. Toxic baits with boric acid (1.0%) mixed in different concentrations of inverted sugar (20%, 30%, and 40%), and hydramethylnon, mixed with sardines (paste), cassava flour and peanut, brown sugar (sucrose), or sardine oil-based gel, were evaluated in a greenhouse and in the field. In the greenhouse experiment, the number of foraging ants was significantly reduced in the pots where the hydramethylnon in sardine paste (Solid S), sardine oil-brown sugar-based gel (GEL SAM), and peanut oil-brown-sugar gel (GEL AM) formulations were applied. The GEL SAM toxic bait effectively reduced the infestation of L. micans, and could be used for indirect control of E. brasiliensis on young grapevines. PMID:27329621

  10. Environmental controls in the water use patterns of a tropical cloud forest tree species, Drimys brasiliensis (Winteraceae).

    PubMed

    Eller, Cleiton B; Burgess, Stephen S O; Oliveira, Rafael S

    2015-04-01

    Trees from tropical montane cloud forest (TMCF) display very dynamic patterns of water use. They are capable of downwards water transport towards the soil during leaf-wetting events, likely a consequence of foliar water uptake (FWU), as well as high rates of night-time transpiration (Enight) during drier nights. These two processes might represent important sources of water losses and gains to the plant, but little is known about the environmental factors controlling these water fluxes. We evaluated how contrasting atmospheric and soil water conditions control diurnal, nocturnal and seasonal dynamics of sap flow in Drimys brasiliensis (Miers), a common Neotropical cloud forest species. We monitored the seasonal variation of soil water content, micrometeorological conditions and sap flow of D. brasiliensis trees in the field during wet and dry seasons. We also conducted a greenhouse experiment exposing D. brasiliensis saplings under contrasting soil water conditions to deuterium-labelled fog water. We found that during the night D. brasiliensis possesses heightened stomatal sensitivity to soil drought and vapour pressure deficit, which reduces night-time water loss. Leaf-wetting events had a strong suppressive effect on tree transpiration (E). Foliar water uptake increased in magnitude with drier soil and during longer leaf-wetting events. The difference between diurnal and nocturnal stomatal behaviour in D. brasiliensis could be attributed to an optimization of carbon gain when leaves are dry, as well as minimization of nocturnal water loss. The leaf-wetting events on the other hand seem important to D. brasiliensis water balance, especially during soil droughts, both by suppressing tree transpiration (E) and as a small additional water supply through FWU. Our results suggest that decreases in leaf-wetting events in TMCF might increase D. brasiliensis water loss and decrease its water gains, which could compromise its ecophysiological performance and survival

  11. Nocardia brasiliensis cell wall lipids modulate macrophage and dendritic responses that favor development of experimental actinomycetoma in BALB/c mice.

    PubMed

    Trevino-Villarreal, J Humberto; Vera-Cabrera, Lucio; Valero-Guillén, Pedro L; Salinas-Carmona, Mario C

    2012-10-01

    Nocardia brasiliensis is a Gram-positive facultative intracellular bacterium frequently isolated from human actinomycetoma. However, the pathogenesis of this infection remains unknown. Here, we used a model of bacterial delipidation with benzine to investigate the role of N. brasiliensis cell wall-associated lipids in experimental actinomycetoma. Delipidation of N. brasiliensis with benzine resulted in complete abolition of actinomycetoma without affecting bacterial viability. Chemical analyses revealed that trehalose dimycolate and an unidentified hydrophobic compound were the principal compounds extracted from N. brasiliensis with benzine. By electron microscopy, the extracted lipids were found to be located in the outermost membrane layer of the N. brasiliensis cell wall. They also appeared to confer acid-fastness. In vitro, the extractable lipids from the N. brasiliensis cell wall induced the production of the proinflammatory cytokines interleukin-1β (IL-1β), IL-6, and CCL-2 in macrophages. The N. brasiliensis cell wall extractable lipids inhibited important macrophage microbicidal effects, such as tumor necrosis factor alpha (TNF-α) and nitric oxide (NO) production, phagocytosis, bacterial killing, and major histocompatibility complex class II (MHC-II) expression in response to gamma interferon (IFN-γ). In dendritic cells (DCs), N. brasiliensis cell wall-associated extractable lipids suppressed MHC-II, CD80, and CD40 expression while inducing tumor growth factor β (TGF-β) production. Immunization with delipidated N. brasiliensis induced partial protection preventing actinomycetoma. These findings suggest that N. brasiliensis cell wall-associated lipids are important for actinomycetoma development by inducing inflammation and modulating the responses of macrophages and DCs to N. brasiliensis. PMID:22851755

  12. Nocardia brasiliensis Cell Wall Lipids Modulate Macrophage and Dendritic Responses That Favor Development of Experimental Actinomycetoma in BALB/c Mice

    PubMed Central

    Trevino-Villarreal, J. Humberto; Vera-Cabrera, Lucio; Valero-Guillén, Pedro L.

    2012-01-01

    Nocardia brasiliensis is a Gram-positive facultative intracellular bacterium frequently isolated from human actinomycetoma. However, the pathogenesis of this infection remains unknown. Here, we used a model of bacterial delipidation with benzine to investigate the role of N. brasiliensis cell wall-associated lipids in experimental actinomycetoma. Delipidation of N. brasiliensis with benzine resulted in complete abolition of actinomycetoma without affecting bacterial viability. Chemical analyses revealed that trehalose dimycolate and an unidentified hydrophobic compound were the principal compounds extracted from N. brasiliensis with benzine. By electron microscopy, the extracted lipids were found to be located in the outermost membrane layer of the N. brasiliensis cell wall. They also appeared to confer acid-fastness. In vitro, the extractable lipids from the N. brasiliensis cell wall induced the production of the proinflammatory cytokines interleukin-1β (IL-1β), IL-6, and CCL-2 in macrophages. The N. brasiliensis cell wall extractable lipids inhibited important macrophage microbicidal effects, such as tumor necrosis factor alpha (TNF-α) and nitric oxide (NO) production, phagocytosis, bacterial killing, and major histocompatibility complex class II (MHC-II) expression in response to gamma interferon (IFN-γ). In dendritic cells (DCs), N. brasiliensis cell wall-associated extractable lipids suppressed MHC-II, CD80, and CD40 expression while inducing tumor growth factor β (TGF-β) production. Immunization with delipidated N. brasiliensis induced partial protection preventing actinomycetoma. These findings suggest that N. brasiliensis cell wall-associated lipids are important for actinomycetoma development by inducing inflammation and modulating the responses of macrophages and DCs to N. brasiliensis. PMID:22851755

  13. [Advances in Molecular Cloning].

    PubMed

    Ashwini, M; Murugan, S B; Balamurugan, S; Sathishkumar, R

    2016-01-01

    "Molecular cloning" meaning creation of recombinant DNA molecules has impelled advancement throughout life sciences. DNA manipulation has become easy due to powerful tools showing exponential growth in applications and sophistication of recombinant DNA technology. Cloning genes has become simple what led to an explosion in the understanding of gene function by seamlessly stitching together multiple DNA fragments or by the use of swappable gene cassettes, maximizing swiftness and litheness. A novel archetype might materialize in the near future with synthetic biology techniques that will facilitate quicker assembly and iteration of DNA clones, accelerating the progress of gene therapy vectors, recombinant protein production processes and new vaccines by in vitro chemical synthesis of any in silico-specified DNA construct. The advent of innovative cloning techniques has opened the door to more refined applications such as identification and mapping of epigenetic modifications and high-throughput assembly of combinatorial libraries. In this review, we will examine the major breakthroughs in cloning techniques and their applications in various areas of biological research that have evolved mainly due to easy construction of novel expression systems. PMID:27028806

  14. Extremal quantum cloning machines

    SciTech Connect

    Chiribella, G.; D'Ariano, G. M.; Perinotti, P.; Cerf, N.J.

    2005-10-15

    We investigate the problem of cloning a set of states that is invariant under the action of an irreducible group representation. We then characterize the cloners that are extremal in the convex set of group covariant cloning machines, among which one can restrict the search for optimal cloners. For a set of states that is invariant under the discrete Weyl-Heisenberg group, we show that all extremal cloners can be unitarily realized using the so-called double-Bell states, whence providing a general proof of the popular ansatz used in the literature for finding optimal cloners in a variety of settings. Our result can also be generalized to continuous-variable optimal cloning in infinite dimensions, where the covariance group is the customary Weyl-Heisenberg group of displacement000.

  15. rPbPga1 from Paracoccidioides brasiliensis Activates Mast Cells and Macrophages via NFkB

    PubMed Central

    Valim, Clarissa Xavier Resende; da Silva, Elaine Zayas Marcelino; Assis, Mariana Aprigio; Fernandes, Fabricio Freitas; Coelho, Paulo Sergio Rodrigues; Oliver, Constance; Jamur, Maria Célia

    2015-01-01

    Background The fungus Paracoccidioides brasiliensis is the leading etiological agent of paracoccidioidomycosis (PCM), a systemic granulomatous disease that typically affects the lungs. Cell wall components of P. brasiliensis interact with host cells and influence the pathogenesis of PCM. In yeast, many glycosylphosphatidylinositol (GPI)-anchored proteins are important in the initial contact with the host, mediating host-yeast interactions that culminate with the disease. PbPga1 is a GPI anchored protein located on the surface of the yeast P. brasiliensis that is recognized by sera from PCM patients. Methodology/Principal Findings Endogenous PbPga1 was localized to the surface of P. brasiliensis yeast cells in the lungs of infected mice using a polyclonal anti-rPbPga1 antibody. Furthermore, macrophages stained with anti-CD38 were associated with P. brasiliensis containing granulomas. Additionally, rPbPga1 activated the transcription factor NFkB in the macrophage cell line Raw 264.7 Luc cells, containing the luciferase gene downstream of the NFkB promoter. After 24 h of incubation with rPbPga1, alveolar macrophages from BALB/c mice were stimulated to release TNF-α, IL-4 and NO. Mast cells, identified by toluidine blue staining, were also associated with P. brasiliensis containing granulomas. Co-culture of P. Brasiliensis yeast cells with RBL-2H3 mast cells induced morphological changes on the surface of the mast cells. Furthermore, RBL-2H3 mast cells were degranulated by P. brasiliensis yeast cells, but not by rPbPga1, as determined by the release of beta-hexosaminidase. However, RBL-2H3 cells activated by rPbPga1 released the inflammatory interleukin IL-6 and also activated the transcription factor NFkB in GFP-reporter mast cells. The transcription factor NFAT was not activated when the mast cells were incubated with rPbPga1. Conclusions/Significance The results indicate that PbPga1 may act as a modulator protein in PCM pathogenesis and serve as a useful target for

  16. Epidemiological and biological aspects on Ornithodoros brasiliensis (mouro tick), an argasidae tick only found on the highlands region of Rio Grande do Sul state, southern Brazil

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The soft tick Ornithodoros brasiliensis (Acari: Argasidae) is present in farms along the highlands of Rio Grande do Sul state in southern Brazil. Reports of human parasitism by O. brasiliensis drew the attention of local health authorities. A preliminary epidemiological survey was conducted to ident...

  17. Type 3 muscarinic receptors contribute to intestinal mucosal homeostasis and clearance of Nippostrongylus brasiliensis through induction of TH2 cytokines.

    PubMed

    McLean, Leon P; Smith, Allen; Cheung, Lumei; Urban, Joseph F; Sun, Rex; Grinchuk, Viktoriya; Desai, Neemesh; Zhao, Aiping; Raufman, Jean-Pierre; Shea-Donohue, Terez

    2016-07-01

    Despite increased appreciation for the role of nicotinic receptors in the modulation of and response to inflammation, the contribution of muscarinic receptors to mucosal homeostasis, clearance of enteric pathogens, and modulation of immune cell function remains relatively undefined. Uninfected and Nippostrongylus brasiliensis-infected wild-type and type 3 muscarinic receptor (M3R)-deficient (Chrm3(-/-)) mice were studied to determine the contribution of M3R to mucosal homeostasis as well as host defense against the TH2-eliciting enteric nematode N. brasiliensis Intestinal permeability and expression of TH1/TH17 cytokines were increased in uninfected Chrm3(-/-) small intestine. Notably, in Chrm3(-/-) mice infected with N. brasiliensis, small intestinal upregulation of TH2 cytokines was attenuated and nematode clearance was delayed. In Chrm3(-/-) mice, TH2-dependent changes in small intestinal function including smooth muscle hypercontractility, increased epithelial permeability, decreased epithelial secretion and absorption, and goblet cell expansion were absent despite N. brasiliensis infection. These findings identify an important role for M3R in host defense and clearance of N. brasiliensis, and support the expanding role of cholinergic muscarinic receptors in maintaining mucosal homeostasis. PMID:27173511

  18. Oenothein B inhibits the expression of PbFKS1 transcript and induces morphological changes in Paracoccidioides brasiliensis.

    PubMed

    Santos, Glaciane D; Ferri, Pedro H; Santos, Suzana C; Bao, Sônia N; Soares, Célia M A; Pereira, Maristela

    2007-11-01

    The fungus Paracoccidioides brasiliensis is the causative agent of paracoccidioidomycosis (PCM), the most prevalent human systemic mycosis in Latin America. Drug toxicity and the appearance of resistant strains have created the need to search for new therapeutic approaches. Plants with reputed antimicrobial properties represent a rich screening source of potential antifungal compounds. In this work, the growth of P. brasiliensis yeast cells was evaluated in the presence of oenothein B extracted from Eugenia uniflora. The oenothein B dosage that most effectively inhibited the development (74%) of P. brasiliensis yeast cells in vitro was 500 microg/ml. To verify if oenothein B interferes with cell morphology, we observed oenothein B-treated yeast cells by electron microscopy. The micrographs showed characteristic cell changes noted with glucan synthesis inhibition, including squashing, rough surface, cell wall rupture and cell membrane recess. The expression of P. brasiliensis genes was evaluated in order to investigate the action of oenothein B. Here we report that oenothein B inhibits 1,3-beta-glucan synthase (PbFKS1) transcript accumulation. The results indicate that oenothein B interferes with the cell morphology of P. brasiliensis, probably by inhibiting the transcription of 1,3-beta-glucan synthase gene, which is involved in the cell wall synthesis. PMID:18033615

  19. Secure the Clones

    NASA Astrophysics Data System (ADS)

    Jensen, Thomas; Kirchner, Florent; Pichardie, David

    Exchanging mutable data objects with untrusted code is a delicate matter because of the risk of creating a data space that is accessible by an attacker. Consequently, secure programming guidelines for Java stress the importance of using defensive copying before accepting or handing out references to an internal mutable object. However, implementation of a copy method (like clone()) is entirely left to the programmer. It may not provide a sufficiently deep copy of an object and is subject to overriding by a malicious sub-class. Currently no language-based mechanism supports secure object cloning. This paper proposes a type-based annotation system for defining modular copy policies for class-based object-oriented programs. A copy policy specifies the maximally allowed sharing between an object and its clone. We present a static enforcement mechanism that will guarantee that all classes fulfill their copy policy, even in the presence of overriding of copy methods, and establish the semantic correctness of the overall approach in Coq. The mechanism has been implemented and experimentally evaluated on clone methods from several Java libraries.

  20. Applications of quantum cloning

    NASA Astrophysics Data System (ADS)

    Pomarico, E.; Sanguinetti, B.; Sekatski, P.; Zbinden, H.; Gisin, N.

    2011-10-01

    Quantum Cloning Machines (QCMs) allow for the copying of information, within the limits imposed by quantum mechanics. These devices are particularly interesting in the high-gain regime, i.e., when one input qubit generates a state of many output qubits. In this regime, they allow for the study of certain aspects of the quantum to classical transition. The understanding of these aspects is the root of the two recent applications that we will review in this paper: the first one is the Quantum Cloning Radiometer, a device which is able to produce an absolute measure of spectral radiance. This device exploits the fact that in the quantum regime information can be copied with only finite fidelity, whereas when a state becomes macroscopic, this fidelity gradually increases to 1. Measuring the fidelity of the cloning operation then allows to precisely determine the absolute spectral radiance of the input optical source. We will then discuss whether a Quantum Cloning Machine could be used to produce a state visible by the naked human eye, and the possibility of a Bell Experiment with humans playing the role of detectors.

  1. The Cloning of America.

    ERIC Educational Resources Information Center

    Dobson, Judith E.; Dobson, Russell L.

    1981-01-01

    Proposes that the U.S. school system purports to prize human variability, but many educators are engaged in activities that seek to homogenize students. Describes these activities, including diagnosis, labeling, ability grouping, and positive reinforcement. Presents suggestions for counselors to combat sources of cloning and self-validation. (RC)

  2. Sequential cloning of chromosomes

    SciTech Connect

    Lacks, S.A.

    1991-12-31

    A method for sequential cloning of chromosomal DNA and chromosomal DNA cloned by this method are disclosed. The method includes the selection of a target organism having a segment of chromosomal DNA to be sequentially cloned. A first DNA segment, having a first restriction enzyme site on either side. homologous to the chromosomal DNA to be sequentially cloned is isolated. A first vector product is formed by ligating the homologous segment into a suitably designed vector. The first vector product is circularly integrated into the target organism`s chromosomal DNA. The resulting integrated chromosomal DNA segment includes the homologous DNA segment at either end of the integrated vector segment. The integrated chromosomal DNA is cleaved with a second restriction enzyme and ligated to form a vector-containing plasmid, which is replicated in a host organism. The replicated plasmid is then cleaved with the first restriction enzyme. Next, a DNA segment containing the vector and a segment of DNA homologous to a distal portion of the previously isolated DNA segment is isolated. This segment is then ligated to form a plasmid which is replicated within a suitable host. This plasmid is then circularly integrated into the target chromosomal DNA. The chromosomal DNA containing the circularly integrated vector is treated with a third, retrorestriction enzyme. The cleaved DNA is ligated to give a plasmid that is used to transform a host permissive for replication of its vector. The sequential cloning process continues by repeated cycles of circular integration and excision. The excision is carried out alternately with the second and third enzymes.

  3. The First Human Cloned Embryo.

    ERIC Educational Resources Information Center

    Cibelli, Jose B.; Lanza, Robert P.; West, Michael D.; Ezzell, Carol

    2002-01-01

    Describes a process known as parthenogenesis which produces cloned, early-stage embryos and human embryos generated only from eggs. Speculates that this technology puts therapeutic cloning within reach. (DDR)

  4. Antibodies Against Glycolipids Enhance Antifungal Activity of Macrophages and Reduce Fungal Burden After Infection with Paracoccidioides brasiliensis.

    PubMed

    Bueno, Renata A; Thomaz, Luciana; Muñoz, Julian E; da Silva, Cássia J; Nosanchuk, Joshua D; Pinto, Márcia R; Travassos, Luiz R; Taborda, Carlos P

    2016-01-01

    Paracoccidioidomycosis is a fungal disease endemic in Latin America. Polyclonal antibodies to acidic glycosphingolipids (GSLs) from Paracoccidioides brasiliensis opsonized yeast forms in vitro increasing phagocytosis and reduced the fungal burden of infected animals. Antibodies to GSL were active in both prophylactic and therapeutic protocols using a murine intratracheal infection model. Pathological examination of the lungs of animals treated with antibodies to GSL showed well-organized granulomas and minimally damaged parenchyma compared to the untreated control. Murine peritoneal macrophages activated by IFN-γ and incubated with antibodies against acidic GSLs more effectively phagocytosed and killed P. brasiliensis yeast cells as well as produced more nitric oxide compared to controls. The present work discloses a novel target of protective antibodies against P. brasiliensis adding to other well-studied mediators of the immune response to this fungus. PMID:26870028

  5. Antibodies Against Glycolipids Enhance Antifungal Activity of Macrophages and Reduce Fungal Burden After Infection with Paracoccidioides brasiliensis

    PubMed Central

    Bueno, Renata A.; Thomaz, Luciana; Muñoz, Julian E.; da Silva, Cássia J.; Nosanchuk, Joshua D.; Pinto, Márcia R.; Travassos, Luiz R.; Taborda, Carlos P.

    2016-01-01

    Paracoccidioidomycosis is a fungal disease endemic in Latin America. Polyclonal antibodies to acidic glycosphingolipids (GSLs) from Paracoccidioides brasiliensis opsonized yeast forms in vitro increasing phagocytosis and reduced the fungal burden of infected animals. Antibodies to GSL were active in both prophylactic and therapeutic protocols using a murine intratracheal infection model. Pathological examination of the lungs of animals treated with antibodies to GSL showed well-organized granulomas and minimally damaged parenchyma compared to the untreated control. Murine peritoneal macrophages activated by IFN-γ and incubated with antibodies against acidic GSLs more effectively phagocytosed and killed P. brasiliensis yeast cells as well as produced more nitric oxide compared to controls. The present work discloses a novel target of protective antibodies against P. brasiliensis adding to other well-studied mediators of the immune response to this fungus. PMID:26870028

  6. Probabilistic Cloning and Quantum Computation

    NASA Astrophysics Data System (ADS)

    Gao, Ting; Yan, Feng-Li; Wang, Zhi-Xi

    2004-06-01

    We discuss the usefulness of quantum cloning and present examples of quantum computation tasks for which the cloning offers an advantage which cannot be matched by any approach that does not resort to quantum cloning. In these quantum computations, we need to distribute quantum information contained in the states about which we have some partial information. To perform quantum computations, we use a state-dependent probabilistic quantum cloning procedure to distribute quantum information in the middle of a quantum computation.

  7. Detection of Antibodies against Paracoccidioides brasiliensis Melanin in In Vitro and In Vivo Studies during Infection ▿

    PubMed Central

    Urán, Martha E.; Nosanchuk, Joshua D.; Restrepo, Angela; Hamilton, Andrew J.; Gómez, Beatriz L.; Cano, Luz E.

    2011-01-01

    Several cell wall constituents, including melanins or melanin-like compounds, have been implicated in the pathogenesis of a wide variety of microbial diseases caused by diverse species of pathogenic bacteria, fungi, and helminthes. Among these microorganisms, the dimorphic fungal pathogen Paracoccidioides brasiliensis produces melanin in its conidial and yeast forms. In the present study, melanin particles from P. brasiliensis were injected into BALB/c mice in order to produce monoclonal antibodies (MAbs). We identified five immunoglobulin G1 (IgG1) κ-chain and four IgM melanin-binding MAbs. The five IgG1 κ-chain isotypes are the first melanin-binding IgG MAbs ever reported. The nine MAbs labeled P. brasiliensis conidia and yeast cells both in vitro and in pulmonary tissues. The MAbs cross-reacted with melanin-like purified particles from other fungi and also with commercial melanins, such as synthetic and Sepia officinalis melanin. Melanization during paracoccidioidomycosis (PCM) was also further supported by the detection of IgG antibodies reactive to melanin from P. brasiliensis conidia and yeast in sera and bronchoalveolar lavage fluids from P. brasiliensis-infected mice, as well as in sera from human patients with PCM. Serum specimens from patients with other mycoses were also tested for melanin-binding antibodies by enzyme-linked immunosorbent assay, and cross-reactivities were detected for melanin particles from different fungal sources. These results suggest that melanin from P. brasiliensis is an immunologically active fungal structure that activates a strong IgG humoral response in humans and mice. PMID:21813659

  8. Detection of Melanin-Like Pigments in the Dimorphic Fungal Pathogen Paracoccidioides brasiliensis In Vitro and during Infection

    PubMed Central

    Gómez, Beatriz L.; Nosanchuk, Joshua D.; Díez, Soraya; Youngchim, Sirida; Aisen, Philip; Cano, Luz E.; Restrepo, Angela; Casadevall, Arturo; Hamilton, Andrew J.

    2001-01-01

    Melanins are implicated in the pathogenesis of several human diseases, including some microbial infections. In this study, we analyzed whether the conidia and the yeasts of the thermally dimorphic fungal pathogen Paracoccidioides brasiliensis produce melanin or melanin-like compounds in vitro and during infection. Growth of P. brasiliensis mycelia on water agar alone produced pigmented conidia, and growth of yeasts in minimal medium with l-3,4-dihydroxyphenylalanine (l-DOPA) produced pigmented cells. Digestion of the pigmented conidia and yeasts with proteolytic enzymes, denaturant, and hot concentrated acid yielded dark particles that were the same size and shape as their propagules. Immunofluorescence analysis demonstrated reactivity of a melanin-binding monoclonal antibody (MAb) with the pigmented conidia, yeasts, and particles. Electron spin resonance spectroscopy identified the yeast-derived particles produced in vitro when P. brasiliensis was grown in l-DOPA medium as a melanin-like compound. Nonreducing polyacrylamide gel electrophoresis of cytoplasmic yeast extract revealed a protein that catalyzed melanin synthesis from l-DOPA. The melanin binding MAb reacted with yeast cells in tissue from mice infected with P. brasiliensis. Finally digestion of infected tissue liberated particles reactive to the melanin binding MAb that had the typical morphology of P. brasiliensis yeasts. These data strongly suggest that P. brasiliensis propagules, both conidia and yeast cells, can produce melanin or melanin-like compounds in vitro and in vivo. Based on what is known about the function of melanin in the virulence of other fungi, this pigment may play a role in the pathogenesis of paracoccidioidomycosis. PMID:11500453

  9. The Homeostasis of Iron, Copper, and Zinc in Paracoccidioides Brasiliensis, Cryptococcus Neoformans Var. Grubii, and Cryptococcus Gattii: A Comparative Analysis

    PubMed Central

    Silva, Mirelle Garcia; Schrank, Augusto; Bailão, Elisa Flávia L.C.; Bailão, Alexandre Melo; Borges, Clayton Luiz; Staats, Charley Christian; Parente, Juliana Alves; Pereira, Maristela; Salem-Izacc, Silvia Maria; Mendes-Giannini, Maria José Soares; Oliveira, Rosely Maria Zancopé; Silva, Lívia Kmetzsch Rosa e; Nosanchuk, Joshua D.; Vainstein, Marilene Henning; de Almeida Soares, Célia Maria

    2011-01-01

    Iron, copper, and zinc are essential for all living organisms. Moreover, the homeostasis of these metals is vital to microorganisms during pathogenic interactions with a host. Most pathogens have developed specific mechanisms for the uptake of micronutrients from their hosts in order to counteract the low availability of essential ions in infected tissues. We report here an analysis of genes potentially involved in iron, copper, and zinc uptake and homeostasis in the fungal pathogens Paracoccidioides brasiliensis, Cryptococcus neoformans var. grubii, and Cryptococcus gattii. Although prior studies have identified certain aspects of metal regulation in Cryptococcus species, little is known regarding the regulation of these elements in P. brasiliensis. We also present amino acid sequences analyses of deduced proteins in order to examine possible conserved domains. The genomic data reveals, for the first time, genes associated to iron, copper, and zinc assimilation and homeostasis in P. brasiliensis. Furthermore, analyses of the three fungal species identified homologs to genes associated with high-affinity uptake systems, vacuolar and mitochondrial iron storage, copper uptake and reduction, and zinc assimilation. However, homologs to genes involved in siderophore production were only found in P. brasiliensis. Interestingly, in silico analysis of the genomes of P. brasiliensis Pb01, Pb03, and Pb18 revealed significant differences in the presence and/or number of genes involved in metal homeostasis, such as in genes related to iron reduction and oxidation. The broad analyses of the genomes of P. brasiliensis, C. neoformans var. grubii, and C. gattii for genes involved in metal homeostasis provide important groundwork for numerous interesting future areas of investigation that are required in order to validate and explore the function of the identified genes and gene pathways. PMID:21833306

  10. A chitinolytic endochitinase and β-N-acetylglucosaminidase-based system from Hevea latex in generating N-acetylglucosamine from chitin.

    PubMed

    Sukprasirt, Pannawich; Wititsuwannakul, Rapepun

    2014-08-01

    An endochitinase and β-N-acetylglucosaminidase (NAGase) were purified and characterised from fresh rubber latex serum. These enzymes were used in a total enzyme-based system to produce pure N-acetylglucosamine (NAG) from chitin. The N-terminal amino acid sequences of both purified endochitinase (KEESRRRRHR) and NAGase (AAVDSDTLEI) lacked homology with other known chitinases, including hevamine from rubber latex lutoids. The apparent kinetic parameters, Km and Vmax, for the endochitinase using 4-MU-β-(NAG)3 as a substrate were 99.73 μM and 29.49 pkat mg(-1), respectively. For NAGase, using 4-MU-β-NAG as a substrate, the corresponding Km and Vmax values were 20.4 μM and 25.82 pkat mg(-1). When an enzyme incubation mixture containing a 1:1 (pkat/pkat) activity mixed ratio of endochitinase: NAGase was employed, the maximum yield of N-acetylglucosamine (NAG) obtained was 98% from β-chitin and 20% from α-chitin. These yields were obtained after 4 days of hydrolysis of equal amounts of β-chitin and α-chitin in the mixture. Thus, β-chitin was the preferred substrate compared to α-chitin by a ratio of nearly five to one. Mass spectroscopic analysis, using electrospray ionisation mass spectrometry (ESI-MS), of the product obtained from β-chitin after digestion (for 24h) depicted one distinct major molecular ion peak m/z 260.1, a small minor ion peak m/z 481.2, a potassium adduct of NAG and a potassium adduct of two NAG molecules. Furthermore, experiments to establish the commercial production of NAG using crude enzymes of Hevea latex serum are currently in progress. PMID:24833032

  11. Diet and helminths of Enyalius brasiliensis (Lacertilia, Iguania, Leiosauridae) in an Atlantic Rainforest remnant in southeastern Brazil.

    PubMed

    Dorigo, T A; Maia-Carneiro, T; Almeida-Gomes, M; Siqueira, C C; Vrcibradic, D; Van Sluys, M; Rocha, C F D

    2014-02-01

    Our study aimed to add information about the diet and endoparasites of Enyalius brasiliensis from an Atlantic Rainforest remnant in the state of Rio de Janeiro, southeastern Brazil. Regarding diet, E. brasiliensis consumed arthropods, with caterpillars and beetles being the most important preys. Regarding helminth parasites, overall prevalence was low (9.5%), with 238 nematodes of the genus Physaloptera found in the stomach of one specimen and one nematode of the genus Rhabdias inside the lung of another. Our results corroborate the observations of previous studies that indicate that lizards of the genus Enyalius tend to feed mainly on relatively large-bodied arthropods and to harbour depauperate endoparasite fauna. PMID:25055102

  12. Recombinational Cloning Using Gateway and In-Fusion Cloning Schemes

    PubMed Central

    Throop, Andrea L.; LaBaer, Joshua

    2015-01-01

    The comprehensive study of protein structure and function, or proteomics, depends on the obtainability of full-length cDNAs in species-specific expression vectors and subsequent functional analysis of the expressed protein. Recombinational cloning is a universal cloning technique based on site-specific recombination that is independent of the insert DNA sequence of interest, which differentiates this method from the classical restriction enzyme-based cloning methods. Recombinational cloning enables rapid and efficient parallel transfer of DNA inserts into multiple expression systems. This unit summarizes strategies for generating expression-ready clones using the most popular recombinational cloning technologies, including the commercially available Gateway® (Life Technologies) and In-Fusion® (Clontech) cloning technologies. PMID:25827088

  13. A conserved dimorphism-regulating histidine kinase controls the dimorphic switching in Paracoccidioides brasiliensis.

    PubMed

    Chaves, Alison F A; Navarro, Marina V; Castilho, Daniele G; Calado, Juliana C P; Conceição, Palloma M; Batista, Wagner L

    2016-08-01

    Paracoccidioides brasiliensis and P. lutzii, thermally dimorphic fungi, are the causative agents of paracoccidioidomycosis (PCM). Paracoccidioides infection occurs when conidia or mycelium fragments are inhaled by the host, which causes the Paracoccidioides cells to transition to the yeast form. The development of disease requires conidia inside the host alveoli to differentiate into yeast cells in a temperature-dependent manner. We describe the presence of a two-component signal transduction system in P. brasiliensis, which we investigated by expression analysis of a hypothetical protein gene (PADG_07579) that showed high similarity with the dimorphism-regulating histidine kinase (DRK1) gene of Blastomyces dermatitidis and Histoplasma capsulatum This gene was sensitive to environmental redox changes, which was demonstrated by a dose-dependent decrease in transcript levels after peroxide stimulation and a subtler decrease in transcript levels after NO stimulation. Furthermore, the higher PbDRK1 levels after treatment with increasing NaCl concentrations suggest that this histidine kinase can play a role as osmosensing. In the mycelium-yeast (M→Y) transition, PbDRK1 mRNA expression increased 14-fold after 24 h incubation at 37°C, consistent with similar observations in other virulent fungi. These results demonstrate that the PbDRK1 gene is differentially expressed during the dimorphic M→Y transition. Finally, when P. brasiliensis mycelium cells were exposed to a histidine kinase inhibitor and incubated at 37°C, there was a delay in the dimorphic M→Y transition, suggesting that histidine kinases could be targets of interest for PCM therapy. PMID:27268997

  14. Bioconcentration and bioaccumulation of metal in freshwater Neotropical fish Geophagus brasiliensis.

    PubMed

    Voigt, Carmen Lúcia; da Silva, Cleber Pinto; Doria, Halina Binde; Randi, Marco Antônio Ferreira; de Oliveira Ribeiro, Ciro Alberto; de Campos, Sandro Xavier

    2015-06-01

    From the concentration in water and sediments, bioconcentration and bioaccumulation of copper (Cu), manganese (Mn), zinc (Zn), iron (Fe), cobalt (Co), cadmium (Cd), chrome (Cr), silver (Ag), lead (Pb), nickel (Ni), aluminum (Al), and arsenic (As) were determined in the gills, liver, and muscles of Geophagus brasiliensis in the Alagados Reservoir, Ponta Grossa, Paraná, Brazil. Metals were quantified through AAS, and a study was carried out on the existing relations between metal and body weight, size, and genre of this species. The level of metal in the water of the reservoir was lower than the maximum set forth in the legislation, except for that of Cd and Fe. In sediments, Cu, Cd, Cr, and Ni presented concentrations above the threshold effect level (TEL). Pb and Cr were above the limits for the G. brasiliensis. The tendency of metals present in the muscles of G. brasiliensis was Al > Cu > Zn > Fe > Co > Mn > Cr > Ag > Ni > Pb > Cd > As. In the gills, it was Al > Fe > Zn > Mn > Co > Ag > Cr > Ni > Cu > As > Pb > Cd, and the liver presented Al > Cu > Zn > Co > Fe > Mn > Pb > Ag > Ni > Cr > As > Cd. The bioconcentration and bioaccumulation of metal in the tissues follow the global tendency liver > gills > muscle. The statistical analysis did not point to significant differences in the metal concentration and body weight, size, and gender of the species in the three tissues under analysis. PMID:25520205

  15. In Vivo Activity of the Benzothiazinones PBTZ169 and BTZ043 against Nocardia brasiliensis

    PubMed Central

    González-Martínez, Norma Alejandra; Lozano-Garza, Hector Gerardo; Castro-Garza, Jorge; De Osio-Cortez, Alexandra; Vargas-Villarreal, Javier; Cavazos-Rocha, Norma; Ocampo-Candiani, Jorge; Makarov, Vadim; Cole, Stewart T.; Vera-Cabrera, Lucio

    2015-01-01

    Background Mycetoma is a neglected, chronic, and deforming infectious disease caused by fungi and actinomycetes. In Mexico, N. brasiliensis is the predominant etiologic agent. Therapeutic alternatives are necessary because the current drug regimens have several disadvantages. Benzothiazinones (BTZ) are a new class of candidate drugs that inhibit decaprenyl-phosphoribose-epimerase (DprE1), an essential enzyme involved in the cell wall biosynthesis of Corynebacterineae. Methodology/Principal findings In this study, the in vitro activity of the next generation BTZ, PBTZ169, was tested against thirty Nocardia brasiliensis isolates. The MIC50 and MIC90 values for PBTZ169 were 0.0075 and 0.03 μg/mL, respectively. Because Nocardia is a potential intracellular bacterium, a THP-1 macrophage monolayer was infected with N. brasiliensis HUJEG-1 and then treated with PBTZ169, resulting in a decrease in the number of colony-forming units (CFUs) at a concentration of 0.25X the in vitro value. The in vivo activity was evaluated after infecting female BALB/c mice in the right hind food-pad. After 6 weeks, treatment was initiated with PBTZ169 and its activity was compared with the first generation compound, BTZ043. Both BTZ compounds were administered at 100 mg/kg twice daily by gavage, and sulfamethoxazole/trimethoprim (SXT), at 100 mg/kg sulfamethoxazole, was used as a positive control. After 22 weeks of therapy, only PBTZ169 and SXT displayed statistically significant activity. Conclusion These results indicate that DprE1 inhibitors may be useful for treating infections of Nocardia and may therefore be active against other actinomycetoma agents. We must test combinations of these compounds with other antimicrobial agents, such as linezolid, tedizolid or SXT, that have good to excellent in vivo activity, as well as new DprE1 inhibitors that can achieve higher plasma levels. PMID:26474057

  16. Overlap extension PCR cloning.

    PubMed

    Bryksin, Anton; Matsumura, Ichiro

    2013-01-01

    Rising demand for recombinant proteins has motivated the development of efficient and reliable cloning methods. Here we show how a beginner can clone virtually any DNA insert into a plasmid of choice without the use of restriction endonucleases or T4 DNA ligase. Chimeric primers encoding plasmid sequence at the 5' ends and insert sequence at the 3' ends are designed and synthesized. Phusion(®) DNA polymerase is utilized to amplify the desired insert by PCR. The double-stranded product is subsequently employed as a pair of mega-primers in a PCR-like reaction with circular plasmids. The original plasmids are then destroyed in restriction digests with Dpn I. The product of the overlap extension PCR is used to transform competent Escherichia coli cells. Phusion(®) DNA polymerase is used for both the amplification and fusion reactions, so both steps can be monitored and optimized in the same way. PMID:23996437

  17. Cloning of murine ferrochelatase.

    PubMed Central

    Brenner, D A; Frasier, F

    1991-01-01

    Ferrochelatase (protoheme ferro-lyase, EC 4.99.1.1) catalyzes the last step in the heme biosynthetic pathway, the chelation of ferrous iron and protoporphyrin to form heme. The activity of ferrochelatase is deficient in the inherited disease protoporphyria. In this study, murine ferrochelatase cDNAs were obtained by screening cDNA libraries with an oligonucleotide probe. The derived amino acid sequence of murine ferrochelatase has 47% identity with the recently cloned Saccharomyces cerevisiae ferrochelatase, but it is not significantly similar to other published sequences. Results of Southern blotting are consistent with a single murine ferrochelatase gene, while Northern blotting demonstrates two ferrochelatase transcripts in all tissues examined. The ferrochelatase protein and mRNAs have different relative concentrations in different tissues. The cloning of murine ferrochelatase cDNAs provides the basis for future studies on ferrochelatase gene expression and on the identification of the molecular defect in protoporphyria. Images PMID:1704134

  18. Morphological Transition of Paracoccidioides brasiliensis Conidia to Yeast Cells: In Vivo Inhibition in Females

    PubMed Central

    Aristizabal, Beatriz H.; Clemons, Karl V.; Stevens, David A.; Restrepo, Angela

    1998-01-01

    Clinical paracoccidioidomycosis is 13 times more common in men than in women. Estrogen inhibits the transition of mycelia or conidia (the saprophytic form of Paracoccidoides brasiliensis) to yeasts (the parasitic form) in vitro. Here, we show that, in male mice that were infected intranasally (mimicking natural infection) the transition of conidia in bronchoalveolar lavage fluids to intermediate forms and yeasts occurred over 24 to 96 h; CFU and yeasts (shown by histopathology) increased subsequently. In females, transition did not occur and infection cleared. These events in vivo are consistent with epidemiological and in vitro observations, suggesting that female hormones block transition and are responsible for resistance. PMID:9784579

  19. Notes on the reproductive biology of the Brazilian electric ray Narcine brasiliensis (Elasmobranchii: Narcinidae).

    PubMed

    Rolim, F A; Rotundo, M M; Vaske-Júnior, T

    2016-07-01

    This study provides information on the reproductive biology of Narcine brasiliensis based on 105 individuals (72 females and 33 males) sampled in São Paulo State, Brazil. The total length at maturity for females was 318·9 mm and for males was 279·8 mm; pregnant females were observed only during summer and autumn. The peak of the gonado-somatic index for females and condition factor for males in the spring suggest a preparation for pregnancy and a mating period during this season. The capture of immature individuals indicates a need for management of the species in this region. PMID:26377171

  20. Data in support of quantitative proteomics to identify potential virulence regulators in Paracoccidioides brasiliensis isolates

    PubMed Central

    Tashima, Alexandre Keiji; Castilho, Daniele Gonçalves; Chaves, Alison Felipe Alencar; Xander, Patricia; Zelanis, André; Batista, Wagner Luiz

    2015-01-01

    Paracoccidioides genus are the etiologic agents of paracoccidioidomycosis (PCM), a systemic mycosis endemic in Latin America. Few virulence factors have been identified in these fungi. This paper describes support data from the quantitative proteomics of Paracoccidioides brasiliensis attenuated and virulent isolates [1]. The protein compositions of two isolates of the Pb18 strain showing distinct infection profiles were quantitatively assessed by stable isotopic dimethyl labeling and proteomic analysis. The mass spectrometry and the analysis dataset have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with identifier PXD000804. PMID:26501084

  1. Cloning-free CRISPR

    PubMed Central

    Arbab, Mandana; Srinivasan, Sharanya; Hashimoto, Tatsunori; Geijsen, Niels; Sherwood, Richard I.

    2015-01-01

    Summary We present self-cloning CRISPR/Cas9 (scCRISPR), a technology that allows for CRISPR/Cas9-mediated genomic mutation and site-specific knockin transgene creation within several hours by circumventing the need to clone a site-specific single-guide RNA (sgRNA) or knockin homology construct for each target locus. We introduce a self-cleaving palindromic sgRNA plasmid and a short double-stranded DNA sequence encoding the desired locus-specific sgRNA into target cells, allowing them to produce a locus-specific sgRNA plasmid through homologous recombination. scCRISPR enables efficient generation of gene knockouts (∼88% mutation rate) at approximately one-sixth the cost of plasmid-based sgRNA construction with only 2 hr of preparation for each targeted site. Additionally, we demonstrate efficient site-specific knockin of GFP transgenes without any plasmid cloning or genome-integrated selection cassette in mouse and human embryonic stem cells (2%–4% knockin rate) through PCR-based addition of short homology arms. scCRISPR substantially lowers the bar on mouse and human transgenesis. PMID:26527385

  2. Sequential cloning of chromosomes

    DOEpatents

    Lacks, Sanford A.

    1995-07-18

    A method for sequential cloning of chromosomal DNA of a target organism is disclosed. A first DNA segment homologous to the chromosomal DNA to be sequentially cloned is isolated. The first segment has a first restriction enzyme site on either side. A first vector product is formed by ligating the homologous segment into a suitably designed vector. The first vector product is circularly integrated into the target organism's chromosomal DNA. The resulting integrated chromosomal DNA segment includes the homologous DNA segment at either end of the integrated vector segment. The integrated chromosomal DNA is cleaved with a second restriction enzyme and ligated to form a vector-containing plasmid, which is replicated in a host organism. The replicated plasmid is then cleaved with the first restriction enzyme. Next, a DNA segment containing the vector and a segment of DNA homologous to a distal portion of the previously isolated DNA segment is isolated. This segment is then ligated to form a plasmid which is replicated within a suitable host. This plasmid is then circularly integrated into the target chromosomal DNA. The chromosomal DNA containing the circularly integrated vector is treated with a third, retrorestriction (class IIS) enzyme. The cleaved DNA is ligated to give a plasmid that is used to transform a host permissive for replication of its vector. The sequential cloning process continues by repeated cycles of circular integration and excision. The excision is carried out alternately with the second and third enzymes.

  3. Sequential cloning of chromosomes

    DOEpatents

    Lacks, S.A.

    1995-07-18

    A method for sequential cloning of chromosomal DNA of a target organism is disclosed. A first DNA segment homologous to the chromosomal DNA to be sequentially cloned is isolated. The first segment has a first restriction enzyme site on either side. A first vector product is formed by ligating the homologous segment into a suitably designed vector. The first vector product is circularly integrated into the target organism`s chromosomal DNA. The resulting integrated chromosomal DNA segment includes the homologous DNA segment at either end of the integrated vector segment. The integrated chromosomal DNA is cleaved with a second restriction enzyme and ligated to form a vector-containing plasmid, which is replicated in a host organism. The replicated plasmid is then cleaved with the first restriction enzyme. Next, a DNA segment containing the vector and a segment of DNA homologous to a distal portion of the previously isolated DNA segment is isolated. This segment is then ligated to form a plasmid which is replicated within a suitable host. This plasmid is then circularly integrated into the target chromosomal DNA. The chromosomal DNA containing the circularly integrated vector is treated with a third, retrorestriction (class IIS) enzyme. The cleaved DNA is ligated to give a plasmid that is used to transform a host permissive for replication of its vector. The sequential cloning process continues by repeated cycles of circular integration and excision. The excision is carried out alternately with the second and third enzymes. 9 figs.

  4. Ethical issues in livestock cloning.

    PubMed

    Thompson, P B

    1999-01-01

    Although cloning may eventually become an important technology for livestock production, four ethical issues must be addressed before the practice becomes widespread. First, researchers must establish that the procedure is not detrimental to the health or well-being of affected animals. Second, animal research institutions should evaluate the net social benefits to livestock producers by weighing the benefits to producers against the opportunity cost of research capacity lost to biomedical projects. Third, scientists should consider the indirect effects of cloning research on the larger ethical issues surrounding human cloning. Finally, the market structure for products of cloned animals should protect individual choice, and should recognize that many individuals find the prospect of cloning (or consuming cloned animals) repugnant. Analysis of these four issues is complicated by spurious arguments alleging that cloning will have a negative impact on environment and genetic diversity. PMID:15719505

  5. Probabilistic cloning of equidistant states

    SciTech Connect

    Jimenez, O.; Roa, Luis; Delgado, A.

    2010-08-15

    We study the probabilistic cloning of equidistant states. These states are such that the inner product between them is a complex constant or its conjugate. Thereby, it is possible to study their cloning in a simple way. In particular, we are interested in the behavior of the cloning probability as a function of the phase of the overlap among the involved states. We show that for certain families of equidistant states Duan and Guo's cloning machine leads to cloning probabilities lower than the optimal unambiguous discrimination probability of equidistant states. We propose an alternative cloning machine whose cloning probability is higher than or equal to the optimal unambiguous discrimination probability for any family of equidistant states. Both machines achieve the same probability for equidistant states whose inner product is a positive real number.

  6. To clone or not to clone--a Jewish perspective.

    PubMed Central

    Lipschutz, J H

    1999-01-01

    Many new reproductive methods such as artificial insemination, in vitro fertilisation, freezing of human embryos, and surrogate motherhood were at first widely condemned but are now seen in Western society as not just ethically and morally acceptable, but beneficial in that they allow otherwise infertile couples to have children. The idea of human cloning was also quickly condemned but debate is now emerging. This article examines cloning from a Jewish perspective and finds evidence to support the view that there is nothing inherently wrong with the idea of human cloning. A hypothesis is also advanced suggesting that even if a body was cloned, the brain, which is the essence of humanity, would remain unique. This author suggests that the debate should be changed from "Is cloning wrong?" to "When is cloning wrong?". PMID:10226913

  7. Systemic increased immune response to Nocardia brasiliensis co-exists with local immunosuppressive microenvironment.

    PubMed

    Salinas-Carmona, Mario Cesar; Rosas-Taraco, Adrian Geovanni; Welsh, Oliverio

    2012-10-01

    Human diseases produced by pathogenic actinomycetes are increasing because they may be present as opportunistic infections. Some of these microbes cause systemic infections associated with immunosuppressive conditions, such as chemotherapy for cancer, immunosuppressive therapy for transplant, autoimmune conditions, and AIDS; while others usually cause localized infection in immunocompetent individuals. Other factors related to this increase in incidence are: antibiotic resistance, not well defined taxonomy, and a delay in isolation and identification of the offending microbe. Examples of these infections are systemic disease and brain abscesses produced by Nocardia asteroides or the located disease by Nocardia brasiliensis, named actinomycetoma. During the Pathogenic Actinomycetes Symposium of the 16th International Symposium on Biology of Actinomycetes (ISBA), held in Puerto Vallarta, Mexico, several authors presented recent research on the mechanisms by which N. brasiliensis modulates the immune system to survive in the host and advances in medical treatment of human actinomycetoma. Antibiotics and antimicrobials that are effective against severe actinomycetoma infections with an excellent therapeutic outcome and experimental studies of drugs that show promising bacterial inhibition in vivo and in vitro were presented. Here we demonstrate a systemic strong acquired immune response in humans and experimental mice at the same time of a local dominance of anti inflammatory cytokines environment. The pathogenic mechanisms of some actinomycetes include generation of an immunosuppressive micro environment to evade the protective immune response. This information will be helpful in understanding pathogenesis and to design new drugs for treatment of actinomycetoma. PMID:22825801

  8. Geographic Discrimination of Paracoccidioides brasiliensis Strains by Randomly Amplified Polymorphic DNA Analysis

    PubMed Central

    Calcagno, Ana María; Niño-Vega, Gustavo; San-Blas, Felipe; San-Blas, Gioconda

    1998-01-01

    Randomly amplified polymorphic DNA (RAPD) analysis of 33 Paracoccidioides brasiliensis strains from Argentina, Brazil, Colombia, Peru, and Venezuela produced reproducible amplification products which were sufficiently polymorphic to allow differentiation of the strains. Types generated with five primers (OPG 03, OPG 05, OPG 14, OPG 16, and OPG 18) resulted in a high discriminatory index (0.956). The discriminatory index was slightly reduced (0.940) when only two primers (OPG 3 and OPG 14) were used. A dendrogram based on these results showed a high degree of similarity among the strains, and genetic differences were expressed in clusters related to geographical regions but not to pathological features of the disease. With a few exceptions, strains were sorted into five groups by geographical origin as follows: group I, Venezuelan strains; group II, Brazilian strains; group III, Peruvian strains; group IV, Colombian strains; and group V, Argentinian strains. The group containing the most disparate strains was group V (discriminatory index, 0.633); the discriminatory index for the other four groups was 0.824. The use of primer OPG 18 by itself was sufficient to discriminate species specificity, and the use of primer OPG 14 by itself was sufficient to discriminate among the geographical locations of the strains in the sample. This method may be helpful for epidemiological studies of P. brasiliensis. PMID:9620409

  9. Monoclonal antibody capture enzyme immunoassay for detection of Paracoccidioides brasiliensis antibodies in paracoccidioidomycosis.

    PubMed Central

    Camargo, Z P; Gesztesi, J L; Saraiva, E C; Taborda, C P; Vicentini, A P; Lopes, J D

    1994-01-01

    Four murine monoclonal antibodies (MAbs 17C, 21A, 21F, and 32B) raised against the 43-kDa glycoprotein of Paracoccidioides brasiliensis were tested in a capture enzyme immunoassay (EIA) for the detection of specific human anti-gp43 immunoglobulin G in patients with paracoccidioidomycosis (PCM). All MAbs reacted similarly in the assay. These MAbs, which detected anti-gp43 at levels of as low as 500 pg/ml, were demonstrated to specifically recognize at least two different epitopes in gp43 binding assays. Specific antibodies in the sera of patients with active PCM were detected at dilutions of as high as 1:819,200, and the reactivities of patient sera, as measured by optical densities, were found to be significantly higher than those of control sera. The comparison between classical ELISA and our capture enzyme immunoassay showed that both sensitivity and specificity were greatly improved by the latter. These MAbs represent the first specific reagents to P. brasiliensis described for use in serological tests for PCM. Images PMID:7814469

  10. Detrimental Effect of Fungal 60-kDa Heat Shock Protein on Experimental Paracoccidioides brasiliensis Infection.

    PubMed

    Fernandes, Fabrício Freitas; Oliveira, Leandro Licursi de; Landgraf, Taise Natali; Peron, Gabriela; Costa, Marcelo Vieira; Coelho-Castelo, Arlete A M; Bonato, Vânia L D; Roque-Barreira, Maria-Cristina; Panunto-Castelo, Ademilson

    2016-01-01

    The genus Paracoccidioides comprises species of dimorphic fungi that cause paracoccidioidomycosis (PCM), a systemic disease prevalent in Latin America. Here, we investigated whether administration of native 60-kDa heat shock protein of P. brasiliensis (nPbHsp60) or its recombinant counterpart (rPbHsp60) affected the course of experimental PCM. Mice were subcutaneously injected with nPbHsp60 or rPbHsp60 emulsified in complete's Freund Adjuvant (CFA) at three weeks after intravenous injection of P. brasiliensis yeasts. Infected control mice were injected with CFA or isotonic saline solution alone. Thirty days after the nPbHsp60 or rPbHsp60 administration, mice showed remarkably increased fungal load, tissue inflammation, and granulomas in the lungs, liver, and spleen compared with control mice. Further, rPbHsp60 treatment (i) decreased the known protective effect of CFA against PCM and (ii) increased the concentrations of IL-17, TNF-α, IL-12, IFN-γ, IL-4, IL-10, and TGF-β in the lungs. Together, our results indicated that PbHsp60 induced a harmful immune response, exacerbated inflammation, and promoted fungal dissemination. Therefore, we propose that PbHsp60 contributes to the fungal pathogenesis. PMID:27598463

  11. [Paracoccidioides brasiliensis infection in a subtropical region with important environmental changes].

    PubMed

    Mangiaterra, M L; Giusiano, G E; Alonso, J M; Gorodner, J O

    1999-07-01

    Paracoccidioidomycosis is a systemic mycoses caused by Paracoccidioides brasiliensis, a dimorphic fungus that infects man through respiratory ducts. It has been accepted that its ecological niche is located in the soil and plants of subtropical forests of Latin America. The Province of Corrientes is located at the northeastern border of Argentina, in a subtropical area where important environmental modifications have been introduced in the last decade as consequence of damming the Paranà river at Yacyretà, one of the biggest hydroelectric dams in the world. Since there are no data on human infection provoqued by this fungal agent in Corrientes, the purpose of this study was to obtain information at present time about infection indexes and to assess if environmental changes introduced in the area could impact on the epidemiology of the disease. Skin tests with paracoccidioidin and histoplasmin were performed on 455 persons of both sexes, from 1 to 73 years of age, who live permanently in the area and voluntarily accepted to be included in the study. Both antigens were employed at the same time in order to evaluate crossed type reactions. Of the 455 persons, 52 (39 males-13 females) were reactive to paracoccidioidin (11.4%), with an increasing prevalence with age. According to previous data, these results would indicate an increase in the index of human infection by P. brasiliensis, and this may be related to the important changes in climatic and environmental conditions introduced in the area in the last years. PMID:10472444

  12. Assessment of water quality and genotoxic impact by toxic metals in Geophagus brasiliensis.

    PubMed

    Morais, Cássio Resende; Carvalho, Stephan Malfitano; Araujo, Galber Rodrigues; Souto, Henrique Nazareth; Bonetti, Ana Maria; Morelli, Sandra; Campos Júnior, Edimar Olegário

    2016-06-01

    This study used the pearl cichlid Geophagus brasiliensis as a bioindicator to survey the health of the aquatic environment on four sites (P1, P2, P3 and P4) of the Mumbuca stream located at Monte Carmelo/MG, Brazil. The selection of different sites was made with reference to the gradient of urban activity and via physicochemical and biological evaluation of water quality and genotoxicity. The water quality index was classified as 'good' for P1 and P4, regular in P2 and 'poor' for P3. The micronuclei (MN) frequency obtained from blood analysis was in agreement with the water quality, such that the higher values of MN were detected in sites evaluated as poor. Water degradation conditions worsen according to the flow of the stream over the sites P1, P2 and P3, but for site P4, located after the Monte Carmelo Sewage Treatment Plant, improvements in the micronuclei frequency are detected. Our results showed high levels of potentially toxic metals (chromium, lead, aluminum and nickel) in specific stream sites (P2 and P3). We suggest that the micronuclei induction in G. brasiliensis could be due to the presence of these compounds. PMID:26991381

  13. Lipidomic Analysis of Extracellular Vesicles from the Pathogenic Phase of Paracoccidioides brasiliensis

    PubMed Central

    Longo, Larissa V. G.; Ganiko, Luciane; Lopes, Felipe G.; Matsuo, Alisson L.; Almeida, Igor C.; Puccia, Rosana

    2012-01-01

    Background Fungal extracellular vesicles are able to cross the cell wall and transport molecules that help in nutrient acquisition, cell defense, and modulation of the host defense machinery. Methodology/Principal Findings Here we present a detailed lipidomic analysis of extracellular vesicles released by Paracoccidioides brasiliensis at the yeast pathogenic phase. We compared data of two representative isolates, Pb3 and Pb18, which have distinct virulence profiles and phylogenetic background. Vesicle lipids were fractionated into different classes and analyzed by either electrospray ionization- or gas chromatography-mass spectrometry. We found two species of monohexosylceramide and 33 phospholipid species, including phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylserine, phosphatidylinositol, and phosphatidylglycerol. Among the phospholipid-bound fatty acids in extracellular vesicles, C181 predominated in Pb3, whereas C18:2 prevailed in Pb18. The prevalent sterol in Pb3 and Pb18 vesicles was brassicasterol, followed by ergosterol and lanosterol. Inter-isolate differences in sterol composition were observed, and also between extracellular vesicles and whole cells. Conclusions/Significance The extensive lipidomic analysis of extracellular vesicles from two P. brasiliensis isolates will help to understand the composition of these fungal components/organelles and will hopefully be useful to study their biogenesis and role in host-pathogen interactions. PMID:22745761

  14. Cell Wall Composition of the Yeast and Mycelial Forms of Paracoccidioides brasiliensis

    PubMed Central

    Kanetsuna, Fuminori; Carbonell, Luis M.; Moreno, Ramon E.; Rodriguez, Joaquin

    1969-01-01

    Isolation and chemical analyses of the cell walls of the yeast (Y form) and mycelial forms (M form) of Paracoccidioides brasiliensis and Blastomyces dermatitidis revealed that their chemical composition is similar and depends on the form. Lipids, chitin, glucans, and proteins are the main constituents of the cell walls of both forms of these fungi. There is no significant difference in the amount of lipids (5 to 10%) and glucans (36 to 47%) contained by the two forms. In both fungi, the Y form has a larger amount of chitin (37 to 48%) than the M form (7 to 18%), whereas the M form has a larger amount of proteins (24 to 41%) than the Y form (7 to 14%). Several properties of the glucan of P. brasiliensis were studied. Almost all of the glucan in the Y form was soluble in 1 n NaOH, was weakly positive in the periodic acid-Schiff reaction, was not hydrolyzed by snail digestive juice, and had α-glycosidic linkage. Glucans of the M form were divided into alkali-soluble (60 to 65%) and alkali-insoluble (35 to 40%) types. The alkali-soluble glucan was similar to that of the Y form; the alkali-insoluble glucan was positive in the periodic acid-Schiff reaction and was hydrolyzed by snail digestive juice. PMID:5776517

  15. DOSE RESPONSE EFFECT OF Paracoccidioides brasiliensis IN AN EXPERIMENTAL MODEL OF ARTHRITIS

    PubMed Central

    Loth, Eduardo Alexandre; Biazim, Samia Khalil; dos Santos, José Henrique Fermino Ferreira; Puccia, Rosana; Brancalhão, Rosimeire Costa; Chasco, Lucinéia de Fátima; Gandra, Rinaldo Ferreira; Simão, Rita de Cássia Garcia; de Franco, Marcello Fabiano

    2014-01-01

    Paracoccidioidomycosis (PCM) is caused by the dimorphic fungus Paracoccidioides brasiliensis (Pb) and corresponds to prevalent systemic mycosis in Latin America. The aim of the present work was to evaluate the dose response effect of the fungal yeast phase for the standardization of an experimental model of septic arthritis. The experiments were performed with groups of 14 rats that received doses of 103, 104 or 105 P. brasiliensis (Pb18) cells. The fungi were injected in 50 µL of phosphate-buffered saline (PBS) directly into the knee joints of the animals. The following parameters were analyzed in this work: the formation of swelling in knees infused with yeast cells and the radiological and anatomopathological alterations, besides antibody titer by ELISA. After 15 days of infection, signs of inflammation were evident. At 45 days, some features of damage and necrosis were observed in the articular cartilage. The systemic dissemination of the fungus was observed in 11% of the inoculated animals, and it was concluded that the experimental model is able to mimic articular PCM in humans and that the dose of 105 yeast cells can be used as standard in this model. PMID:24879005

  16. Rediscovering hermaphroditism in Grammatidae with the description of the testicular gland in Brazilian Basslet Gramma brasiliensis.

    PubMed

    Leite, J R; Freitas, M O; Sanches, E G; Gomes, M L M; Hostim-Silva, M; Cole, K S

    2016-04-19

    Many aspects of sex change in reef fishes have been studied, including behavior and social organization. However, gonad histology remains the most robust way to identify sexual patterns in fishes. Some uncommon tissues remain poorly described, such as the accessory gonadal structures found in species from the Gobiidae family, which are rare in other bony fishes. This is the first report of the testicular gland in Gramma brasiliensis and for the Grammatidae family. Between April 2011 and February 2012 eighty specimens were collected during four dive campaigns on the Taipus de Fora reef (13°56'20"S 38°55'32"W), Bahia, Northeast Brazil, and their sex was determined. Thirteen per cent of the active-females and 90% of the active-males had testicular gland tissue in their ovotestis. This discovery led to additional research into the characteristics of the gland tissue and its relationship with gonadal maturation. Three patterns of testicular gland development were found in Brazilian basslet ovotestis. Both ova and sperm-producing gonad contained testicular gland tissue, and the appearance of this tissue seems to be the first modification of ovotestis tissue marking the beginning of the protogynous sex-change process in G. brasiliensis. PMID:27097082

  17. Enzyme-linked immunosorbent assay for serological diagnosis of Nocardia brasiliensis and clinical correlation with mycetoma infections.

    PubMed Central

    Salinas-Carmona, M C; Welsh, O; Casillas, S M

    1993-01-01

    We previously identified three immunodominant antigens obtained from a Nocardia brasiliensis cell extract and recognized by sera from mycetoma patients (M. C. Salinas-Carmona, L. Vera, O. Welsh, and M. Rodríguez, Zentralbl. Bakteriol. 276:390-397, 1992). In the present work, we obtained a crude extract from a mass culture of N. brasiliensis HUJEG-1 and purified two immunodominant antigens, the 26- and 24-kDa proteins, by using simple physiochemical techniques. With these antigens, we developed a conventional solid-phase enzyme-linked immunosorbent assay and tested 30 serum samples from mycetoma patients, 29 from tuberculosis patients, 24 from a leprosy group, and 31 from healthy individuals. Our results show for the first time statistically significant differences in serology among these groups. All mycetoma patients with a positive culture for N. brasiliensis had absorbance values higher than 0.3. On the other hand, the mycobacterium-infected patients as well as the healthy individuals all had absorbance values below that level. Moreover, we found a close correlation between the clinical condition of the mycetoma patients and the anti-26- and anti-24-kDa protein antibody concentrations. We therefore propose the use of this assay in routine clinical laboratories to confirm the diagnosis of N. brasiliensis infection in human mycetoma cases. In addition, the possible application of this assay in the serodiagnosis of Nocardia asteroides infection is also discussed. Images PMID:8263174

  18. Pseudozyma brasiliensis sp. nov., a xylanolytic, ustilaginomycetous yeast species isolated from an insect pest of sugarcane roots.

    PubMed

    Oliveira, Juliana Velasco de Castro; Borges, Thuanny A; Corrêa dos Santos, Renato Augusto; Freitas, Larissa F D; Rosa, Carlos Augusto; Goldman, Gustavo Henrique; Riaño-Pachón, Diego Mauricio

    2014-06-01

    A novel ustilaginomycetous yeast isolated from the intestinal tract of an insect pest of sugarcane roots in Ribeirão Preto, São Paulo State, Brazil, represents a novel species of the genus Pseudozyma based on molecular analyses of the D1/D2 rDNA large subunit and the internal transcribed spacer (ITS1+ITS2) regions. The name Pseudozyma brasiliensis sp. nov. is proposed for this species, with GHG001(T) ( = CBS 13268(T) = UFMG-CM-Y307(T)) as the type strain. P. brasiliensis sp. nov. is a sister species of Pseudozyma vetiver, originally isolated from leaves of vetiver grass and sugarcane in Thailand. P. brasiliensis sp. nov. is able to grow well with xylan as the sole carbon source and produces high levels of an endo-1,4-xylanase that has a higher specific activity in comparison with other eukaryotic xylanases. This enzyme has a variety of industrial applications, indicating the great biotechnological potential of P. brasiliensis. PMID:24682702

  19. [Biology of Agistemus brasiliensis Matioli, Ueckermann & Oliveira (Acari: Stigmaeidae) and its predation potential on Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae)].

    PubMed

    Matioli, André L; de Oliveira, Carlos A L

    2007-01-01

    The present work aimed to study the biology of Agistemus brasiliensis Matioli, Ueckermann & Oliveira at the following temperatures, namely 15, 20, 25, 30 and 35 degrees C, fed with Brevipalpus phoenicis (Geijskes) and Typha pollen, in laboratory conditions. Life tables were calculated to evaluate the biological parameters. The optimal development of A. brasiliensis took place at 29 degrees C. The values of T (time of generation - days), R0 and r m at 30 degrees C were, 13.95, 16.25 and 0.20, respectively. The prey consumption of A. brasiliensis was studied at the densities of 5, 10, 20, 40 and 60 leprosis mite females per cage (3 cm in diameter) on citrus fruits at 29 degrees C. The maximum prey, namely 7.6 B. phoenicis females per day, were consumed at a density of 20 leprosis mites. At densities of above 40 leprosis mites per cage, A. brasiliensis oviposits 4.7 eggs per day, in comparison to 2.5 eggs per day at 20 mites per cage. PMID:17934624

  20. Vector capacity of members of Triatoma brasiliensis species complex: The need to extend Chagas disease surveillance to Triatoma melanica.

    PubMed

    Folly-Ramos, Elaine; Dornak, L Lynnette; Orsolon, Guilherme; Gonçalves, Teresa Cristina Monte; Lilioso, Mauricio; Costa, Jane; Almeida, Carlos Eduardo

    2016-06-01

    We conducted a lab-based comparative study on vector capacity features of two species of triatomines: Triatoma brasiliensis and T. melanica. Both are members of the T. brasiliensis species complex. The former is the most important Chagas disease vector in the northeastern region of Brazil. To date, no transmission via T. melanica has been recorded. Immature insects exhibited distinct intermoult periods without a direct relationship to a given species. Females of T. brasiliensis consumed an average of 1.9 times more meals (mean = 12.92 vs 6.63) and survived for a shorter period (mean =330.8 days) than T. melanica (mean = 365.2 days), probably due to the cost of reproduction (all significant at P<0.05). These data support the idea that T. brasiliensis is more adapted to lab conditions and is more able to infest domiciles than T. melanica. We also found significant distinctions in other features between these species, such as the elapsed time without eating before molting, which was higher for the second, third, and fifth nymph stages of T. melanica. Regarding features analyzed related to vector capacity, insects of all life stages of both species were considered competent to transmit Trypanosoma cruzi because they needed many feedings (mean =1.5-3.82) to moult and because a high proportion (>39%) of insects defecated rapidly (<30 s) after feeding. Overall, results highlight the need to extend vector surveillance to T. melanica. PMID:27232124

  1. Polyketide Synthase Gene Diversity within the Microbiome of the Sponge Arenosclera brasiliensis, Endemic to the Southern Atlantic Ocean

    PubMed Central

    Trindade-Silva, Amaro E.; Rua, Cintia P. J.; Andrade, Bruno G. N.; Vicente, Ana Carolina Paulo; Silva, Genivaldo G. Z.

    2013-01-01

    Microbes associated with marine sponges are considered important producers of bioactive, structurally unique polyketides. The synthesis of such secondary metabolites involves type I polyketide synthases (PKSs), which are enzymes that reach a maximum complexity degree in bacteria. The Haplosclerida sponge Arenosclera brasiliensis hosts a complex microbiota and is the source of arenosclerins, alkaloids with cytotoxic and antibacterial activity. In the present investigation, we performed high-throughput sequencing of the ketosynthase (KS) amplicon to investigate the diversity of PKS genes present in the metagenome of A. brasiliensis. Almost 4,000 ketosynthase reads were recovered, with about 90% annotated automatically as bacterial. A total of 235 bacterial KS contigs was rigorously assembled from this sequence pool and submitted to phylogenetic analysis. A great diversity of six type I PKS groups has been consistently detected in our phylogenetic reconstructions, including a novel and A. brasiliensis-exclusive group. Our study is the first to reveal the diversity of type I PKS genes in A. brasiliensis as well as the potential of its microbiome to serve as a source of new polyketides. PMID:23275501

  2. Thermal preferences and limits of Triatoma brasiliensis in its natural environment--field observations while host searching.

    PubMed

    Catalá, Silvia; Bezerra, Claudia Mendonça; Diotaiuti, Lileia

    2015-09-01

    The goal of this work was to explore the thermal relationship between foraging Triatoma brasiliensis and its natural habitat during the hottest season in the state of Ceará, Brazil. The thermal profiles were determined using infrared analysis. Although the daily temperature of rock surfaces varied in a wide range, T. brasiliensis selected to walk through areas with temperatures between 31.7-40.5ºC. The temperature of T. brasiliensis body surface ranged from 32.8-34.4ºC, being higher in legs than the abdomen. A strong relationship was found between the temperature of the insect and the temperature of rock crevices where they were hidden (r: 0.96, p < 0.05). The species was active at full sunlight being a clear example of how the light-dark rhythm may be altered, even under predation risk. Our results strongly suggest a thermal borderline for T. brasiliensis foraging activity near 40ºC. The simultaneous determination of insect body and rock temperatures here presented are the only obtained in natural habitats for this or other triatomines. PMID:26517659

  3. Chemical and agronomic development of Kalanchoe brasiliensis Camb. and Kalanchoe pinnata Pers under light and temperature levels.

    PubMed

    Cruz, Bruna P; Chedier, Luciana M; Fabri, Rodrigo L; Pimenta, Daniel S

    2011-12-01

    This study compares the development of Kalanchoe brasiliensis and Kalanchoe pinnata, which are medicinal species known as "saião" and "folha da fortuna" that are used interchangeably by the population for medicinal purposes. The experiment consisted of 20 plots/species planted in plastic bags with homogeneous substrate in a randomized design, which grown under light levels (25%, 50%, 70%, full sunlight) at environment temperature, and a treatment under a plastic with greater temperature range than the external environment. It was obtained for K. pinnata a greater plant height, total length of sprouts, stems, production and dry matter content of leaves than that obtained for K. brasiliensis, which achieved higher averages only for the length of lateral branches. The species showed increases in height, which varied in inverse proportion to the light, and it was observed the influence of temperature in K. pinnata. The production and dry matter content of leaves were proportional to the luminosity; the same occurred in the thickness of leaves for K. brasiliensis. In the swelling index and Brix degree, K. brasiliensis showed higher averages than K. pinnata. In relation to the total content of flavonoids it was not observed significant differences for both species. The analyzed parameters showed the main differences in the agronomic development of the two species. PMID:22146966

  4. cDNA encoding a polypeptide including a hevein sequence

    DOEpatents

    Raikhel, Natasha V.; Broekaert, Willem F.; Chua, Nam-Hai; Kush, Anil

    1993-02-16

    A cDNA clone (HEV1) encoding hevein was isolated via polymerase chain reaction (PCR) using mixed oligonucleotides corresponding to two regions of hevein as primers and a Hevea brasiliensis latex cDNA library as a template. HEV1 is 1018 nucleotides long and includes an open reading frame of 204 amino acids. The deduced amino acid sequence contains a pu GOVERNMENT RIGHTS This application was funded under Department of Energy Contract DE-AC02-76ER01338. The U.S. Government has certain rights under this application and any patent issuing thereon.

  5. Therapeutic cloning and reproductive liberty.

    PubMed

    Sparrow, Robert

    2009-04-01

    Concern for "reproductive liberty" suggests that decisions about embryos should normally be made by the persons who would be the genetic parents of the child that would be brought into existence if the embryo were brought to term. Therapeutic cloning would involve creating and destroying an embryo, which, if brought to term, would be the offspring of the genetic parents of the person undergoing therapy. I argue that central arguments in debates about parenthood and genetics therefore suggest that therapeutic cloning would be prima facie unethical unless it occurred with the consent of the parents of the person being cloned. Alternatively, if therapeutic cloning is thought to be legitimate, this undermines the case for some uses of reproductive cloning by implying that the genetic relation it establishes between clones and DNA donors does not carry the same moral weight as it does in cases of normal reproduction. PMID:19240247

  6. Domestic, peridomestic and wild hosts in the transmission of Trypanosoma cruzi in the Caatinga area colonised by Triatoma brasiliensis.

    PubMed

    Bezerra, Claudia Mendonça; Cavalcanti, Luciano Pamplona de Góes; Souza, Rita de Cássia Moreira de; Barbosa, Silvia Ermelinda; Xavier, Samanta Cristina das Chagas; Jansen, Ana Maria; Ramalho, Relrison Dias; Diotaiut, Liléia

    2014-08-22

    The role played by different mammal species in the maintenance of Trypanosoma cruzi is not constant and varies in time and place. This study aimed to characterise the importance of domestic, wild and peridomestic hosts in the transmission of T. cruzi in Tauá, state of Ceará, Caatinga area, Brazil, with an emphasis on those environments colonised by Triatoma brasiliensis. Direct parasitological examinations were performed on insects and mammals, serologic tests were performed on household and outdoor mammals and multiplex polymerase chain reaction was used on wild mammals. Cytochrome b was used as a food source for wild insects. The serum prevalence in dogs was 38% (20/53), while in pigs it was 6% (2/34). The percentages of the most abundantly infected wild animals were as follows: Thrichomys laurentius 74% (83/112) and Kerodon rupestris 10% (11/112). Of the 749 triatomines collected in the household research, 49.3% (369/749) were positive for T. brasiliensis, while 6.8% were infected with T. cruzi (25/369). In captured animals, T. brasiliensis shares a natural environment with T. laurentius, K. rupestris, Didelphis albiventris, Monodelphis domestica, Galea spixii, Wiedomys pyrrhorhinos, Conepatus semistriatus and Mus musculus. In animals identified via their food source, T. brasiliensis shares a natural environment with G. spixii, K. rupestris, Capra hircus, Gallus gallus, Tropidurus oreadicus and Tupinambis merianae. The high prevalence of T. cruzi in household and peridomiciliar animals reinforces the narrow relationship between the enzootic cycle and humans in environments with T. brasiliensis and characterises it as ubiquitous. PMID:25165976

  7. Domestic, peridomestic and wild hosts in the transmission of Trypanosoma cruzi in the Caatinga area colonised by Triatoma brasiliensis

    PubMed Central

    Bezerra, Claudia Mendonça; Cavalcanti, Luciano Pamplona de Góes; de Souza, Rita de Cássia Moreira; Barbosa, Silvia Ermelinda; Xavier, Samanta Cristina das Chagas; Jansen, Ana Maria; Ramalho, Relrison Dias; Diotaiut, Liléia

    2014-01-01

    The role played by different mammal species in the maintenance of Trypanosoma cruzi is not constant and varies in time and place. This study aimed to characterise the importance of domestic, wild and peridomestic hosts in the transmission of T. cruzi in Tauá, state of Ceará, Caatinga area, Brazil, with an emphasis on those environments colonised by Triatoma brasiliensis. Direct parasitological examinations were performed on insects and mammals, serologic tests were performed on household and outdoor mammals and multiplex polymerase chain reaction was used on wild mammals. Cytochrome b was used as a food source for wild insects. The serum prevalence in dogs was 38% (20/53), while in pigs it was 6% (2/34). The percentages of the most abundantly infected wild animals were as follows: Thrichomys laurentius 74% (83/112) and Kerodon rupestris 10% (11/112). Of the 749 triatomines collected in the household research, 49.3% (369/749) were positive for T. brasiliensis, while 6.8% were infected with T. cruzi (25/369). In captured animals, T. brasiliensis shares a natural environment with T. laurentius, K. rupestris, Didelphis albiventris, Monodelphis domestica, Galea spixii, Wiedomys pyrrhorhinos, Conepatus semistriatus and Mus musculus. In animals identified via their food source, T. brasiliensis shares a natural environment with G. spixii, K. rupestris, Capra hircus, Gallus gallus, Tropidurus oreadicus and Tupinambis merianae. The high prevalence of T. cruzi in household and peridomiciliar animals reinforces the narrow relationship between the enzootic cycle and humans in environments with T. brasiliensis and characterises it as ubiquitous. PMID:25410992

  8. Domestic, peridomestic and wild hosts in the transmission of Trypanosoma cruzi in the Caatinga area colonised by Triatoma brasiliensis.

    PubMed

    Bezerra, Claudia Mendonça; Cavalcanti, Luciano Pamplona de Góes; Souza, Rita de Cássia Moreira de; Barbosa, Silvia Ermelinda; Xavier, Samanta Cristina das Chagas; Jansen, Ana Maria; Ramalho, Relrison Dias; Diotaiut, Liléia

    2014-11-01

    The role played by different mammal species in the maintenance of Trypanosoma cruzi is not constant and varies in time and place. This study aimed to characterise the importance of domestic, wild and peridomestic hosts in the transmission of T. cruzi in Tauá, state of Ceará, Caatinga area, Brazil, with an emphasis on those environments colonised by Triatoma brasiliensis. Direct parasitological examinations were performed on insects and mammals, serologic tests were performed on household and outdoor mammals and multiplex polymerase chain reaction was used on wild mammals. Cytochrome b was used as a food source for wild insects. The serum prevalence in dogs was 38% (20/53), while in pigs it was 6% (2/34). The percentages of the most abundantly infected wild animals were as follows: Thrichomys laurentius 74% (83/112) and Kerodon rupestris 10% (11/112). Of the 749 triatomines collected in the household research, 49.3% (369/749) were positive for T. brasiliensis, while 6.8% were infected with T. cruzi (25/369). In captured animals, T. brasiliensis shares a natural environment with T. laurentius, K. rupestris, Didelphis albiventris, Monodelphis domestica, Galea spixii, Wiedomys pyrrhorhinos, Conepatus semistriatus and Mus musculus. In animals identified via their food source, T. brasiliensis shares a natural environment with G. spixii, K. rupestris, Capra hircus, Gallus gallus, Tropidurus oreadicus and Tupinambis merianae. The high prevalence of T. cruzi in household and peridomiciliar animals reinforces the narrow relationship between the enzootic cycle and humans in environments with T. brasiliensis and characterises it as ubiquitous. PMID:25410992

  9. Self-Cloning CRISPR.

    PubMed

    Arbab, Mandana; Sherwood, Richard I

    2016-01-01

    CRISPR/Cas9-gene editing has emerged as a revolutionary technology to easily modify specific genomic loci by designing complementary sgRNA sequences and introducing these into cells along with Cas9. Self-cloning CRISPR/Cas9 (scCRISPR) uses a self-cleaving palindromic sgRNA plasmid (sgPal) that recombines with short PCR-amplified site-specific sgRNA sequences within the target cell by homologous recombination to circumvent the process of sgRNA plasmid construction. Through this mechanism, scCRISPR enables gene editing within 2 hr once sgRNA oligos are available, with high efficiency equivalent to conventional sgRNA targeting: >90% gene knockout in both mouse and human embryonic stem cells and cancer cell lines. Furthermore, using PCR-based addition of short homology arms, we achieve efficient site-specific knock-in of transgenes such as GFP without traditional plasmid cloning or genome-integrated selection cassette (2% to 4% knock-in rate). The methods in this paper describe the most rapid and efficient means of CRISPR gene editing. © 2016 by John Wiley & Sons, Inc. PMID:27532819

  10. Therapeutic cloning: The ethical limits

    SciTech Connect

    Whittaker, Peter A. . E-mail: p.whittaker@lancaster.ac.uk

    2005-09-01

    A brief outline of stem cells, stem cell therapy and therapeutic cloning is given. The position of therapeutic cloning with regard to other embryonic manipulations - IVF-based reproduction, embryonic stem formation from IVF embryos and reproductive cloning - is indicated. The main ethically challenging stages in therapeutic cloning are considered to be the nuclear transfer process including the source of eggs for this and the destruction of an embryo to provide stem cells for therapeutic use. The extremely polarised nature of the debate regarding the status of an early human embryo is noted, and some potential alternative strategies for preparing immunocompatible pluripotent stem cells are indicated.

  11. Human cloning and child welfare.

    PubMed Central

    Burley, J; Harris, J

    1999-01-01

    In this paper we discuss an objection to human cloning which appeals to the welfare of the child. This objection varies according to the sort of harm it is expected the clone will suffer. The three formulations of it that we will consider are: 1. Clones will be harmed by the fearful or prejudicial attitudes people may have about or towards them (H1); 2. Clones will be harmed by the demands and expectations of parents or genotype donors (H2); 3. Clones will be harmed by their own awareness of their origins, for example the knowledge that the genetic donor is a stranger (H3). We will show why these three versions of the child welfare objection do not necessarily supply compelling reasons to ban human reproductive cloning. The claim that we will develop and defend in the course of our discussion is that even if it is the case that a cloned child will suffer harms of the type H1-H3, it is none the less permissible to conceive by cloning so long as these cloning-induced welfare deficits are not such as to blight the existence of the resultant child, whoever this may be. PMID:10226914

  12. Molecular Cloning of HbPR-1 Gene from Rubber Tree, Expression of HbPR-1 Gene in Nicotiana benthamiana and Its Inhibition of Phytophthora palmivora

    PubMed Central

    Khunjan, Uraiwan; Ekchaweng, Kitiya; Panrat, Tanate; Tian, Miaoying; Churngchow, Nunta

    2016-01-01

    This is the first report to present a full-length cDNA (designated HbPR-1) encoding a putative basic HbPR-1 protein from rubber tree (Hevea brasiliensis) treated with salicylic acid. It was characterized and also expressed in Nicotiana benthamiana using Agrobacterium-mediated transient gene expression system in order to investigate the role of HbPR-1 gene in rubber tree against its oomycete pathogen Phytopthora palmivora and to produce recombinant HbPR-1 protein for microbial inhibition test. The HbPR-1 cDNA was 647 bp long and contained an open reading frame of 492 nucleotides encoding 163 amino acid residues with a predicted molecular mass of 17,681 Da and an isoelectric point (pI) of 8.56, demonstrating that HbPR-1 protein belongs to the basic PR-1 type. The predicted 3D structure of HbPR-1 was composed of four α-helices, three β-sheets, seven strands, and one junction loop. Expression and purification of recombinant HbPR-1 protein were successful using Agrobacterium-mediated transient expression and one-step of affinity chromatography. Heterologous expression of HbPR-1 in N. benthamiana reduced necrosis areas which were inoculated with P. palmivora zoospores, indicating that the expressed HbPR-1 protein played an important role in plant resistance to pathogens. The purified recombinant HbPR-1 protein was found to inhibit 64% of P. palmivora zoospore germination on a water agar plate compared with control, suggesting that it was an antimicrobial protein against P. palmivora. PMID:27337148

  13. Molecular Cloning of HbPR-1 Gene from Rubber Tree, Expression of HbPR-1 Gene in Nicotiana benthamiana and Its Inhibition of Phytophthora palmivora.

    PubMed

    Khunjan, Uraiwan; Ekchaweng, Kitiya; Panrat, Tanate; Tian, Miaoying; Churngchow, Nunta

    2016-01-01

    This is the first report to present a full-length cDNA (designated HbPR-1) encoding a putative basic HbPR-1 protein from rubber tree (Hevea brasiliensis) treated with salicylic acid. It was characterized and also expressed in Nicotiana benthamiana using Agrobacterium-mediated transient gene expression system in order to investigate the role of HbPR-1 gene in rubber tree against its oomycete pathogen Phytopthora palmivora and to produce recombinant HbPR-1 protein for microbial inhibition test. The HbPR-1 cDNA was 647 bp long and contained an open reading frame of 492 nucleotides encoding 163 amino acid residues with a predicted molecular mass of 17,681 Da and an isoelectric point (pI) of 8.56, demonstrating that HbPR-1 protein belongs to the basic PR-1 type. The predicted 3D structure of HbPR-1 was composed of four α-helices, three β-sheets, seven strands, and one junction loop. Expression and purification of recombinant HbPR-1 protein were successful using Agrobacterium-mediated transient expression and one-step of affinity chromatography. Heterologous expression of HbPR-1 in N. benthamiana reduced necrosis areas which were inoculated with P. palmivora zoospores, indicating that the expressed HbPR-1 protein played an important role in plant resistance to pathogens. The purified recombinant HbPR-1 protein was found to inhibit 64% of P. palmivora zoospore germination on a water agar plate compared with control, suggesting that it was an antimicrobial protein against P. palmivora. PMID:27337148

  14. Morphological differentiation and function of the coelomocytes in the parasitic stages of Nippostrongylus brasiliensis.

    PubMed

    Weinstein, Paul P

    2006-10-01

    Female and male worms of Nippostrongylus brasiliensis exhibited sexual dimorphism based on the number of coelomocytes present. A surprising multiplicity of diverse morphological types of coelomocytes developed in both female and male worms during the parasitic cycle. Cytoplasmic processes began to appear on the surface membrane of coelomocytes in the late third-stage larvae (L3s) in the lungs, and they increased greatly in type, size, and morphology during the fourth and fifth stages. These structures were characterized primarily as complex filopodia, pseudopodia, and cytoplasmic pearls, which resulted in the formation of highly pleomorphic cells. Pearls, starting as small protuberances, progressively increased in size and number with larval growth and development. In the adult worms, a novel process of autocannibalism was initiated in many of the very large coelomocytes. The pearls grew enormously in size at the expense of the cytoplasm, forming a peripheral garland in 1 plane surrounding a residual, small, flat, cytoplasmic core containing the nucleus. The underlying "strategy" was to increase the surface-to-volume ratio of these huge cells to overcome the restriction imposed by eutely; the coelomocytes do not undergo cell division. This morphological innovation makes possible a more efficient uptake of nutrients and exocytosis of waste matter. Vesicles (presumably lysosomes) in the coelomocytes of the infective L3 store an extraordinarily high concentration of vitamin B12 (cobalamin, Cbl). At present, the only physiological function that can be assigned to coelomocytes of N. brasiliensis is the uptake, concentration, and storage of Cbl in the free-living stages, with the subsequent release of the molecule from the vesicles in the early phase of parasitism. Thus, stored Cbl in the infective L3 is made available for biochemical processes during the critical period of larval growth and differentiation initiated in the lung. A model of a hypothetical coelomocyte is

  15. Further drimane sesquiterpenes from Drimys brasiliensis stem barks with cytotoxic potential.

    PubMed

    Fratoni, Eduarda; Claudino, Vanessa Duarte; Yunes, Rosendo Augusto; Franchi, Gilberto C; Nowill, Alexandre E; Filho, Valdir Cechinel; Monache, Franco Delle; Malheiros, Angela

    2016-07-01

    Drimys brasiliensis Miers (Winteraceae) is used in folk medicine for the treatment of cancer. Its anti-tumor activity has been demonstrated in vitro models using extracts and isolated compounds. This study investigates the cytotoxic effects of stem bark extracts of D. brasiliensis as well as isolated compounds that may be responsible for the activitys and evaluates them in leukemia cells. The stem bark extract were subjected to column chromatography, and the structures of compounds were elucidated based on spectroscopic methods by using NMR and infrared spectroscopy and GC/MS. The cytotoxicity of the isolated compounds was evaluated in chronic myeloid (K562) and acute B lymphoblastic (Nalm6) leukemia cells using tetrazolium assay (MTT). Two new compounds were isolated 1β-O-p-methoxy-E-cinnamoyl-5α-keto-11α-enol-albicanol (1a) and the isomer 1β-O-p-methoxy-E-cinnamoyl-5α-keto-11β-enol-albicanol (1b) and 1β-O-p-methoxy-E-cinnamoyl-isodrimeninol (2). The known compounds polygonal acid (3a) and the isomer isopolygonal acid (3b), fuegin (4a) and the isomer epifuegin (4b), the mixture drimanial (5) and 1β-O-(p-methoxy-E-cinnamoyl)-6α-hydroxypolygodial (6) were also isolated. The drimanes (1-4) and drimanial (5), 1β-(p-coumaroyloxy)-polygodial (7), 1β-(p-methoxycinnamoyl)-polygodial (8), and polygodial (9) isolated previously were assessed in tumor cells. The IC50 values were between 3.56 and 128.91 μM. 1-β-(p-cumaroiloxi)-polygodial showed the best result with IC50 8.18 and 3.56 μM by K562 and Nalm6, respectively. The chloroform extract of the stem bark of D. brasiliensis is a great source of drimane sesquiterpenes. Our experimental data suggest that drimanes are responsible for cytotoxicity activity demonstrated by this species, especially those with the aldehyde group linked to carbons C-11 and C-12. PMID:27095358

  16. [The discrete horror of cloning].

    PubMed

    Guibourg, Ricardo A

    2009-01-01

    The author raises the topic of cloning after the decision of the Argentine government, which concerned for the "dignity of the human person", passed a decree of need and urgency, No. 200/97 (Annex), prohibiting cloning experiments with human beings. Therefore, considering that the topic is so terribly urgent and necessary, the author feels it is timely to consider it. PMID:19860340

  17. Animal Cloning and Food Safety

    MedlinePlus

    ... from clones and their offspring out of the food chain until CVM could further evaluate the issue. back to top FDA Studies Cloning For more than five years, CVM ... evaluate the safety of food from these animals. The resulting report, called a ...

  18. CATO: The Clone Alignment Tool.

    PubMed

    Henstock, Peter V; LaPan, Peter

    2016-01-01

    High-throughput cloning efforts produce large numbers of sequences that need to be aligned, edited, compared with reference sequences, and organized as files and selected clones. Different pieces of software are typically required to perform each of these tasks. We have designed a single piece of software, CATO, the Clone Alignment Tool, that allows a user to align, evaluate, edit, and select clone sequences based on comparisons to reference sequences. The input and output are designed to be compatible with standard data formats, and thus suitable for integration into a clone processing pipeline. CATO provides both sequence alignment and visualizations to facilitate the analysis of cloning experiments. The alignment algorithm matches each of the relevant candidate sequences against each reference sequence. The visualization portion displays three levels of matching: 1) a top-level summary of the top candidate sequences aligned to each reference sequence, 2) a focused alignment view with the nucleotides of matched sequences displayed against one reference sequence, and 3) a pair-wise alignment of a single reference and candidate sequence pair. Users can select the minimum matching criteria for valid clones, edit or swap reference sequences, and export the results to a summary file as part of the high-throughput cloning workflow. PMID:27459605

  19. CATO: The Clone Alignment Tool

    PubMed Central

    Henstock, Peter V.; LaPan, Peter

    2016-01-01

    High-throughput cloning efforts produce large numbers of sequences that need to be aligned, edited, compared with reference sequences, and organized as files and selected clones. Different pieces of software are typically required to perform each of these tasks. We have designed a single piece of software, CATO, the Clone Alignment Tool, that allows a user to align, evaluate, edit, and select clone sequences based on comparisons to reference sequences. The input and output are designed to be compatible with standard data formats, and thus suitable for integration into a clone processing pipeline. CATO provides both sequence alignment and visualizations to facilitate the analysis of cloning experiments. The alignment algorithm matches each of the relevant candidate sequences against each reference sequence. The visualization portion displays three levels of matching: 1) a top-level summary of the top candidate sequences aligned to each reference sequence, 2) a focused alignment view with the nucleotides of matched sequences displayed against one reference sequence, and 3) a pair-wise alignment of a single reference and candidate sequence pair. Users can select the minimum matching criteria for valid clones, edit or swap reference sequences, and export the results to a summary file as part of the high-throughput cloning workflow. PMID:27459605

  20. South American Leaf Blight of the Rubber Tree (Hevea spp.): New Steps in Plant Domestication using Physiological Features and Molecular Markers

    PubMed Central

    Lieberei, Reinhard

    2007-01-01

    Background Rubber trees (Hevea spp.) are perennial crops of Amazonian origin that have been spread over the whole tropical belt to guarantee worldwide production of natural rubber. This crop plant has found its place in many national economies of producing countries, and although its domestication by selection of suitable genotypes was very slow, it contributes a lot to the welfare of small farmers worldwide. Its development is limited by severe diseases. In South America, the main fungal disease of rubber trees is the South American leaf blight (SALB) caused by the ascomycete Microcyclus ulei. This fungus inhibits natural rubber production on a commercial scale in South and Central America. Scope The disease is still restricted to its continent of origin, but its potential to be distributed around the world rises with every transcontinental airline connection that directly links tropical regions. The need to develop control measures against the disease is an urgent task and must be carried out on an international scale. All control efforts so far taken since 1910 have ended in a miserable failure. Even the use of modern systemic fungicides and use of greatly improved application techniques have failed to prevent large losses and dieback of trees. The results of research dealing with both the disease and the pathosystem over more than 50 years are summarized and placed into perspective. Future Prospects A detailed knowledge of this host–pathogen combination requires understanding of the dynamics of Hevea leaf development, the biochemical potential for cyanide liberation, and molecular data for several types of resistance factors. Resolution of the Hevea–SALB problem may serve as a model for future host–pathogen studies of perennial plants requiring a holistic approach. PMID:17650512

  1. Therapeutic cloning: promises and issues

    PubMed Central

    Kfoury, Charlotte

    2007-01-01

    Advances in biotechnology necessitate both an understanding of scientific principles and ethical implications to be clinically applicable in medicine. In this regard, therapeutic cloning offers significant potential in regenerative medicine by circumventing immunorejection, and in the cure of genetic disorders when used in conjunction with gene therapy. Therapeutic cloning in the context of cell replacement therapy holds a huge potential for de novo organogenesis and the permanent treatment of Parkinson’s disease, Duchenne muscular dystrophy, and diabetes mellitus as shown by in vivo studies. Scientific roadblocks impeding advancement in therapeutic cloning are tumorigenicity, epigenetic reprogramming, mitochondrial heteroplasmy, interspecies pathogen transfer, low oocyte availability. Therapeutic cloning is also often tied to ethical considerations concerning the source, destruction and moral status of IVF embryos based on the argument of potential. Legislative and funding issues are also addressed. Future considerations would include a distinction between therapeutic and reproductive cloning in legislative formulations. PMID:18523539

  2. Isolation and preliminary characterization of auxotrophic and morphological mutants of the yeastlike form of Paracoccidioides brasiliensis.

    PubMed Central

    San Blas, F; Centeno, S

    1977-01-01

    N-methyl-N'-nitro-N-nitrosoguanidine, which is known to be a very effective mutagen in many systems, was used to induce mutants in the yeastlike form of Paracoccidioides brasiliensis strain IVIC Pb9, an imperfect fungus. Forty-three auxotrophic and 27 prototrophic morphological mutants were isolated after treatment with 50 mug of nitrosoguanidine per ml in 0.1 M citrate buffer, pH 5.0. Auxotrophic mutants required primarily either amino acids, purines, or pyrimidines. Some auxotrophs were also morphological mutants. The main morphological difference from the parental strain was the texture or the color of the yeast-like colonies. Only one prototrophic morphological mutant differed in the size and form of the yeastlike cells when compared with the parental strain. Suxotrophic mutants were used in pairwise combination to attempt heterokaryon formation without success. Images PMID:830638

  3. Structure and ultrastructure of leaf and calyx glands in Galphimia brasiliensis (Malpighiaceae).

    PubMed

    Castro, M A; Vega, A S; Múlgura, M E

    2001-11-01

    The present study describes the anatomical structure of calyx and leaf glands in Galphimia brasiliensis and analyzes the mechanism of secretion. The glands are marginal and suprabasal, cup-shaped, sessile, and scarcely visible with the naked eye. Light microscopy reveals the following features: a thin, smooth cuticle; unistratified secretory cells; subglandular parenchyma; and vascular bundle supply composed of phloem and xylem with abundant druses of calcium oxalate. Transmission electron microscopy reveals the presence of secretory cells with conspicuous nuclei, dense cytoplasm, lipid droplets, numerous vesicles, mitochondria, Golgi, rough endoplasmic reticulum (RER), and elongated plastids with osmiophilic contents. The secretion reaches the apoplastic space and accumulates beneath the cuticle. Finally, the viscous, translucent exudate is eliminated by mechanical rupture of the cuticle. Histochemical analysis confirms that lipids are the main constituent. Small amounts of polysaccharides were also identified. PMID:21669626

  4. Tourism values for Mexican free-tailed bat (Tadarida brasiliensis mexicana) viewing

    USGS Publications Warehouse

    Bagstad, Kenneth J.; Widerholdt, Ruscena

    2013-01-01

    Migratory species provide diverse ecosystem services to people, but these values have seldom been estimated rangewide for a single species. In this article, we summarize visitation and consumer surplus for recreational visitors to viewing sites for the Mexican free-tailed bat (Tadarida brasiliensis mexicana) throughout the Southwestern United States. Public bat viewing opportunities are available at 17 of 25 major roosts across six states; on an annual basis, we estimate that over 242,000 visitors view bats, gaining over $6.5 million in consumer surplus. A better understanding of spatial mismatches between the areas where bats provide value to people and areas most critical for maintaining migratory populations can better inform conservation planning, including economic incentive systems for conservation.

  5. Factors associated with Paracoccidiodes brasiliensis infection among permanent residents of three endemic areas in Colombia.

    PubMed Central

    Cadavid, D.; Restrepo, A.

    1993-01-01

    The natural habitat of Paracoccidioides brasiliensis, the aetiologic agent of paracoccidioidomycosis, has not been determined. Consequently, the events leading to the acquisition of infection remain controversial. To identify factors associated with infection in endemic areas we conducted a survey in three rural communities in Colombia where we had previously diagnosed paracoccidioidomycosis in children. Permanent residents were surveyed taking into consideration environmental and occupational variables. Skin tests were used to classify subjects as infected or non-infected. Variables found associated with infection were: (i) community A: previous residence around Porce river and agriculture in vegetable gardens; (ii) community C: frequent use of specific water sources; (iii) community V: housekeeping activities, and (iv) total group: age > 25 years and contact with bats. Residents in communities with higher prevalence of infection were older, had more complex residence history, and referred more contact with armadillos than residents of communities with lower infection. PMID:8348926

  6. Induction of micronuclei by alkaloids extracted from Senecio brasiliensis and stored for 23 years.

    PubMed

    Santos-Mello, R; Deimlimg, L I; Lauer Júnior, C M; Almeida, A

    2002-04-26

    In the present study, we report the results of an investigation on pyrrolizidine alkaloids extracted from Senecio brasiliensis (Sprengel) Less., which were stored for more than 23 years under variable conditions of temperature and humidity and exposed to light. Both the crude alkaloid (integerrimine+retrorsine+impurities) and pure integerrimine conserved the ability to induce acute toxicity in mice, leading to the death of the animals in less than 24h. The alkaloids also conserved the potential to induce significant increases in micronucleus frequencies in polychromatic erythrocytes of mouse bone marrow compared to the negative control. The administration of alkaloids to lymphocyte cultures blocked with cytochalasin-B showed no significant increase in micronucleus frequency in binucleated cells, probably due to the lack of a metabolic activation mechanism. However, an antimitotic effect was observed. PMID:11943607

  7. Selenoprotein Expression in Macrophages Is Critical for Optimal Clearance of Parasitic Helminth Nippostrongylus brasiliensis.

    PubMed

    Nelson, Shakira M; Shay, Ashley E; James, Jamaal L; Carlson, Bradley A; Urban, Joseph F; Prabhu, K Sandeep

    2016-02-01

    The plasticity of macrophages is evident in helminthic parasite infections, providing protection from inflammation. Previously we demonstrated that the micronutrient selenium induces a phenotypic switch in macrophage activation from a classically activated (pro-inflammatory; M1/CAM) toward an alternatively activated (anti-inflammatory; M2/AAM) phenotype, where cyclooxygenase (COX)-dependent cyclopentenone prostaglandin J2 (15d-PGJ2) plays a key role. Here, we hypothesize that dietary selenium modulates macrophage polarization toward an AAM phenotype to assist in the increasing clearance of adult Nippostrongylus brasiliensis, a gastrointestinal nematode parasite. Mice on a selenium-adequate (0.08 ppm) diet significantly augmented intestinal AAM presence while decreasing adult worms and fecal egg production when compared with infection of mice on selenium-deficient (<0.01 ppm) diet. Further increase in dietary selenium to supraphysiological levels (0.4 ppm) had very little or no impact on worm expulsion. Normal adult worm clearance and enhanced AAM marker expression were observed in the selenium-supplemented Trsp(fl/fl)Cre(WT) mice that express selenoproteins driven by tRNA(Sec) (Trsp), whereas N. brasiliensis-infected Trsp(fl/fl)Cre(LysM) selenium-supplemented mice showed a decreased clearance, with lowered intestinal expression of several AAM markers. Inhibition of the COX pathway with indomethacin resulted in delayed worm expulsion in selenium-adequate mice. This was rescued with 15d-PGJ2, which partially recapitulated the effect of selenium supplementation on fecal egg output in addition to increasing markers of AAMs in the small intestine. Antagonism of PPARγ blocked the effect of selenium. These results suggest that optimal expression of selenoproteins and selenium-dependent production of COX-derived endogenous prostanoids, such as Δ(12)-PGJ2 and 15d-PGJ2, may regulate AAM activation to enhance anti-helminthic parasite responses. PMID:26644468

  8. Human cloning: can it be made safe?

    PubMed

    Rhind, Susan M; Taylor, Jane E; De Sousa, Paul A; King, Tim J; McGarry, Michelle; Wilmut, Ian

    2003-11-01

    There are continued claims of attempts to clone humans using nuclear transfer, despite the serious problems that have been encountered in cloning other mammals. It is known that epigenetic and genetic mechanisms are involved in clone failure, but we still do not know exactly how. Human reproductive cloning is unethical, but the production of cells from cloned embryos could offer many potential benefits. So, can human cloning be made safe? PMID:14634633

  9. The rubber tree genome shows expansion of gene family associated with rubber biosynthesis.

    PubMed

    Lau, Nyok-Sean; Makita, Yuko; Kawashima, Mika; Taylor, Todd D; Kondo, Shinji; Othman, Ahmad Sofiman; Shu-Chien, Alexander Chong; Matsui, Minami

    2016-01-01

    Hevea brasiliensis Muell. Arg, a member of the family Euphorbiaceae, is the sole natural resource exploited for commercial production of high-quality natural rubber. The properties of natural rubber latex are almost irreplaceable by synthetic counterparts for many industrial applications. A paucity of knowledge on the molecular mechanisms of rubber biosynthesis in high yield traits still persists. Here we report the comprehensive genome-wide analysis of the widely planted H. brasiliensis clone, RRIM 600. The genome was assembled based on ~155-fold combined coverage with Illumina and PacBio sequence data and has a total length of 1.55 Gb with 72.5% comprising repetitive DNA sequences. A total of 84,440 high-confidence protein-coding genes were predicted. Comparative genomic analysis revealed strong synteny between H. brasiliensis and other Euphorbiaceae genomes. Our data suggest that H. brasiliensis's capacity to produce high levels of latex can be attributed to the expansion of rubber biosynthesis-related genes in its genome and the high expression of these genes in latex. Using cap analysis gene expression data, we illustrate the tissue-specific transcription profiles of rubber biosynthesis-related genes, revealing alternative means of transcriptional regulation. Our study adds to the understanding of H. brasiliensis biology and provides valuable genomic resources for future agronomic-related improvement of the rubber tree. PMID:27339202

  10. Seamless Ligation Cloning Extract (SLiCE) cloning method.

    PubMed

    Zhang, Yongwei; Werling, Uwe; Edelmann, Winfried

    2014-01-01

    SLiCE (Seamless Ligation Cloning Extract) is a novel cloning method that utilizes easy to generate bacterial cell extracts to assemble multiple DNA fragments into recombinant DNA molecules in a single in vitro recombination reaction. SLiCE overcomes the sequence limitations of traditional cloning methods, facilitates seamless cloning by recombining short end homologies (15-52 bp) with or without flanking heterologous sequences and provides an effective strategy for directional subcloning of DNA fragments from bacterial artificial chromosomes or other sources. SLiCE is highly cost-effective and demonstrates the versatility as a number of standard laboratory bacterial strains can serve as sources for SLiCE extract. We established a DH10B-derived E. coli strain expressing an optimized λ prophage Red recombination system, termed PPY, which facilitates SLiCE with very high efficiencies. PMID:24395368

  11. Methylotroph cloning vehicle

    DOEpatents

    Hanson, Richard S.; Allen, Larry N.

    1989-04-25

    A cloning vehicle comprising: a replication determinant effective for replicating the vehicle in a non-C.sub.1 -utilizing host and in a C.sub.1 -utilizing host; DNA effective to allow the vehicle to be mobilized from the non-C.sub.1 -utilizing host to the C.sub.1 -utilizing host; DNA providing resistance to two antibiotics to which the wild-type C.sub.1 -utilizing host is susceptible, each of the antibiotic resistance markers having a recognition site for a restriction endonuclease; a cos site; and a means for preventing replication in the C.sub.1 -utilizing host. The vehicle is used for complementation mapping as follows. DNA comprising a gene from the C.sub.1 -utilizing organism is inserted at the restriction nuclease recognition site, inactivating the antibiotic resistance marker at that site. The vehicle can then be used to form a cosmid structure to infect the non-C.sub.1 -utilizing (e.g., E. coli) host, and then conjugated with a selected C.sub.1 -utilizing mutant. Resistance to the other antibiotic by the mutant is a marker of the conjugation. Other phenotypical changes in the mutant, e.g., loss of an auxotrophic trait, is attributed to the C.sub.1 gene. The vector is also used to inactivate genes whose protein products catalyze side reactions that divert compounds from a biosynthetic pathway to a desired product, thereby producing an organism that makes the desired product in higher yields.

  12. Nitric Oxide Participation in the Fungicidal Mechanism of Gamma Interferon-Activated Murine Macrophages against Paracoccidioides brasiliensis Conidia

    PubMed Central

    Gonzalez, Angel; de Gregori, Waldemar; Velez, Diana; Restrepo, Angela; Cano, Luz E.

    2000-01-01

    Paracoccidioidomycosis, a systemic mycosis restricted to Latin America and produced by the dimorphic fungus Paracoccidioides brasiliensis, is probably acquired by inhalation of conidia produced by the mycelial form. The macrophage (Mφ) represents the major cell defense against this pathogen; when activated with gamma interferon (IFN-γ), murine Mφs kill the fungus by an oxygen-independent mechanism. Our goal was to determine the role of nitric oxide in the fungicidal effect of Mφs on P. brasiliensis conidia. The results revealed that IFN-γ-activated murine Mφs inhibited the conidium-to-yeast transformation process in a dose-dependent manner; maximal inhibition was observed in Mφs activated with 50 U/ml and incubated for 96 h at 37°C. When Mφs were activated with 150 to 200 U of cytokine per ml, the number of CFU was 70% lower than in nonactivated controls, indicating that there was a fungicidal effect. The inhibitory effect was reversed by the addition of anti-IFN-γ monoclonal antibodies. Activation by IFN-γ also enhanced Mφ nitric oxide production, as revealed by increasing NO2 values (8 ± 3 μM in nonactivated Mφs versus 43 ± 13 μM in activated Mφs). The neutralization of IFN-γ also reversed nitric oxide production at basal levels (8 ± 5 μM). Additionally, we found that there was a significant inverse correlation (r = −0.8975) between NO2− concentration and transformation of P. brasiliensis conidia. Additionally, treatment with any of the three different nitric oxide inhibitors used (arginase, NG-monomethyl-l-arginine, and aminoguanidine), reverted the inhibition of the transformation process with 40 to 70% of intracellular yeast and significantly reduced nitric oxide production. These results show that IFN-γ-activated murine Mφs kill P. brasiliensis conidia through the l-arginine–nitric oxide pathway. PMID:10768942

  13. Distributional potential of the Triatoma brasiliensis species complex at present and under scenarios of future climate conditions

    PubMed Central

    2014-01-01

    Background The Triatoma brasiliensis complex is a monophyletic group, comprising three species, one of which includes two subspecific taxa, distributed across 12 Brazilian states, in the caatinga and cerrado biomes. Members of the complex are diverse in terms of epidemiological importance, morphology, biology, ecology, and genetics. Triatoma b. brasiliensis is the most disease-relevant member of the complex in terms of epidemiology, extensive distribution, broad feeding preferences, broad ecological distribution, and high rates of infection with Trypanosoma cruzi; consequently, it is considered the principal vector of Chagas disease in northeastern Brazil. Methods We used ecological niche models to estimate potential distributions of all members of the complex, and evaluated the potential for suitable adjacent areas to be colonized; we also present first evaluations of potential for climate change-mediated distributional shifts. Models were developed using the GARP and Maxent algorithms. Results Models for three members of the complex (T. b. brasiliensis, N = 332; T. b. macromelasoma, N = 35; and T. juazeirensis, N = 78) had significant distributional predictivity; however, models for T. sherlocki and T. melanica, both with very small sample sizes (N = 7), did not yield predictions that performed better than random. Model projections onto future-climate scenarios indicated little broad-scale potential for change in the potential distribution of the complex through 2050. Conclusions This study suggests that T. b. brasiliensis is the member of the complex with the greatest distributional potential to colonize new areas: overall; however, the distribution of the complex appears relatively stable. These analyses offer key information to guide proactive monitoring and remediation activities to reduce risk of Chagas disease transmission. PMID:24886587

  14. Miltefosine is active against Sporothrix brasiliensis isolates with in vitro low susceptibility to amphotericin B or itraconazole.

    PubMed

    Borba-Santos, Luana Pereira; Gagini, Thalita; Ishida, Kelly; de Souza, Wanderley; Rozental, Sonia

    2015-04-01

    Sporotrichosis is a common mycosis caused by dimorphic fungi from the Sporothrix schenckii complex. In recent years, sporotrichosis incidence rates have increased in the Brazilian state of Rio de Janeiro, where Sporothrix brasiliensis is the species more frequently isolated from patients. The standard antifungals itraconazole and amphotericin B are recommended as first-line therapy for cutaneous/lymphocutaneous and disseminated sporotrichosis, respectively, although decreased sensitivity to these drugs in vitro was reported for clinical isolates of S. brasiliensis. Here, we evaluated the activity of the phospholipid analogue miltefosine - already in clinical use against leishmaniasis - towards the pathogenic yeast form of S. brasiliensis isolates with low sensitivity to itraconazole or amphotericin B in vitro. Miltefosine had fungicidal activity, with minimum inhibitory concentration (MIC) values of 1-2 µg ml(-1). Miltefosine exposure led to loss of plasma membrane integrity, and transmission electron microscopy (TEM) analysis revealed a decrease in cytoplasmic electron density, alterations in the thickness of cell wall layers and accumulation of an electron-dense material in the cell wall. Flow cytometry analysis using an anti-melanin antibody revealed an increase in cell wall melanin in yeasts treated with miltefosine, when compared with control cells. The cytotoxicity of miltefosine was comparable to those of amphotericin B, but miltefosine showed a higher selectivity index towards the fungus. Our results suggest that miltefosine could be an effective alternative for the treatment of S. brasiliensis sporotrichosis, when standard treatment fails. Nevertheless, in vivo studies are required to confirm the antifungal potential of miltefosine for the treatment of sporotrichosis. PMID:25681323

  15. Antifungal activity of schinol and a new biphenyl compound isolated from Schinus terebinthifolius against the pathogenic fungus Paracoccidioides brasiliensis

    PubMed Central

    2010-01-01

    Background The aim of this study was to isolate and identify the antifungal compounds from the extracts of Schinus terebinthifolius (Anacardiaceae) against clinical isolates of the pathogenic fungus Paracoccidioides brasiliensis. Methods The hexane and dichlomethane fractions from leaves and stems of S. terebinthifolius were fractionated using several chromatography techniques to afford four compounds. Results The compounds isolated from S. terebinthifolius were identified as schinol (1), a new biphenyl compound, namely, 4'-ethyl-4-methyl-2,2',6,6'-tetrahydroxy[1,1'-biphenyl]-4,4'-dicarboxylate (2), quercetin (3), and kaempferol (4). Compounds 1 and 2 were active against different strains of P. brasiliensis, showing a minimal inhibitory concentration value against the isolate Pb B339 of 15.6 μg/ml. The isolate Pb 1578 was more sensitive to compound 1 with a MIC value of 7.5 μg/ml. Schinol presented synergistic effect only when combined with itraconazole. The compounds isolated from S. terebinthifolius were not able to inhibit cell wall synthesis or assembly using the sorbitol assay. Conclusion This work reveals for the first time the occurrence of compound 2 and discloses activity of compounds 1 and 2 against several clinical isolates of P. brasiliensis. These results justify further studies to clarify the mechanisms of action of these compounds. PMID:20939907

  16. Lesions associated with Halocercus brasiliensis Lins de Almeida, 1933 in the lungs of dolphins stranded in the Northeast of Brazil.

    PubMed

    Guimarães, J P; Febronio, A M B; Vergara-Parente, J E; Werneck, M R

    2015-04-01

    The parasitic fauna of cetaceans is an important tool for ecological studies, including analyses on the causes of death. Halocercus brasiliensis is a nematode frequently found in the bronchi and bronchioles of some cetaceans, and it is commonly associated with focal inflammation of the respiratory tract leading to bacterial pneumonia and septicemia and, sometimes, to death. The objective of this study was to report infections by H. brasiliensis in the respiratory tract of Delphinidae stranded on the northern seaside of Bahia, Sergipe, and south of Alagoas, all states in the northeast region of Brazil. A total of 30 individuals, 1 Feresa attenuate (pygmy killer whale), 9 Stenella clymene (Clymene dolphin), and 20 Sotalia guianensis (Guiana dolphin) were studied. In 16 of them, the presence of H. brasiliensis was observed with a mean intensity of 3.5 ± 0.6 (range 1-9) in the hosts. Macroscopically, parasitic calcified nodules, lung congestion, edema, and emphysema were observed. Histopathological examination showed interstitial and granulomatous pneumonia with multifocal infiltrates, discrete to moderate edema, congestion, diffuse hemorrhage, and foci of calcification. We conclude that parasitic pneumonia in the sampled individuals may have directly contributed to stranding and death of the animals. PMID:25171594

  17. Effect of energetic cost to maintain the trap for Myrmeleon brasiliensis (Neuroptera, Myrmeleontidae) in its development and adult size.

    PubMed

    Lima, T N; Silva, D C R

    2016-07-25

    Antlion larvae Myrmeleon brasiliensis Návas, 1914 (Neuroptera, Myrmeleontidae) are sit-and-wait predators who build traps to catch their prey. The aim of this study was to observe under laboratory conditions, how the energy cost spent on maintenance of their traps affects: the larval developmental time, time spent as a pupa, mortality rate of larvae and adult size. M. brasiliensis larvae were collected in the municipality of Aquidauana, Mato Grosso do Sul, Brazil and were individually maintained in plastic containers and subjected to two treatments. In the control treatment larvae did not have their traps disturbed while in the manipulated treatment, larvae had their traps disturbed three times a week. The experiments were followed until adult emergence. When the adults emerged, their body size (head-abdomen), anterior and posterior wing span and width were measured. Furthermore, the number of larvae that died during the experiment was recorded. The results showed that the larvae whose traps were manipulated had longer larval development time, smaller pupal development time and were smaller adults. It can be concluded that the energy expenditure spent on maintenance of the trap constructed by M. brasiliensis larvae can affect the development of negative ways, represented by a longer larval development and reduced adult size. PMID:27463831

  18. Cloning of a quantum measurement

    SciTech Connect

    Bisio, Alessandro; D'Ariano, Giacomo Mauro; Perinotti, Paolo; Sedlak, Michal

    2011-10-15

    We analyze quantum algorithms for cloning of a quantum measurement. Our aim is to mimic two uses of a device performing an unknown von Neumann measurement with a single use of the device. When the unknown device has to be used before the bipartite state to be measured is available we talk about 1{yields}2 learning of the measurement, otherwise the task is called 1{yields}2 cloning of a measurement. We perform the optimization for both learning and cloning for arbitrary dimension d of the Hilbert space. For 1{yields}2 cloning we also propose a simple quantum network that achieves the optimal fidelity. The optimal fidelity for 1{yields}2 learning just slightly outperforms the estimate and prepare strategy in which one first estimates the unknown measurement and depending on the result suitably prepares the duplicate.

  19. A Clone of Your Own.

    ERIC Educational Resources Information Center

    Bilodeau, Kirsten

    1997-01-01

    Describes an activity used at the Washington Park Arboretum that helps students understand cloning through plant propagation. Students also learn how to make a pot from recycled newspapers and how to make soil that is appropriate for the plants. (DDR)

  20. Human Cloning: Let's Discuss It.

    ERIC Educational Resources Information Center

    Taras, Loretta; Stavroulakis, Anthea M.; Ortiz, Mary T.

    1999-01-01

    Describes experiences with holding discussions on cloning at a variety of levels in undergraduate biology courses. Discusses teaching methods used and student reactions to the discussions. Contains 12 references. (WRM)

  1. Methylotroph cloning vehicle

    DOEpatents

    Hanson, R.S.; Allen, L.N.

    1989-04-25

    A cloning vehicle comprising: a replication determinant effective for replicating the vehicle in a non-C[sub 1]-utilizing host and in a C[sub 1]-utilizing host; DNA effective to allow the vehicle to be mobilized from the non-C[sub 1]-utilizing host to the C[sub 1]-utilizing host; DNA providing resistance to two antibiotics to which the wild-type C[sub 1]-utilizing host is susceptible, each of the antibiotic resistance markers having a recognition site for a restriction endonuclease; a cos site; and a means for preventing replication in the C[sub 1]-utilizing host. The vehicle is used for complementation mapping as follows. DNA comprising a gene from the C[sub 1]-utilizing organism is inserted at the restriction nuclease recognition site, inactivating the antibiotic resistance marker at that site. The vehicle can then be used to form a cosmid structure to infect the non-C[sub 1]-utilizing (e.g., E. coli) host, and then conjugated with a selected C[sub 1]-utilizing mutant. Resistance to the other antibiotic by the mutant is a marker of the conjugation. Other phenotypical changes in the mutant, e.g., loss of an auxotrophic trait, is attributed to the C[sub 1] gene. The vector is also used to inactivate genes whose protein products catalyze side reactions that divert compounds from a biosynthetic pathway to a desired product, thereby producing an organism that makes the desired product in higher yields. 3 figs.

  2. Local cloning of entangled qubits

    SciTech Connect

    Choudhary, Sujit K.; Kunkri, Samir; Rahaman, Ramij; Roy, Anirban

    2007-11-15

    We discuss the exact cloning of orthogonal but entangled qubits under local operations and classical communication. The amount of entanglement necessary in a blank copy is obtained for various cases. Surprisingly, this amount is more than 1 ebit for certain sets of two nonmaximal but equally entangled states of two qubits. To clone any three Bell states, at least log{sub 2} 3 ebit is necessary.

  3. Artificial cloning of domestic animals.

    PubMed

    Keefer, Carol L

    2015-07-21

    Domestic animals can be cloned using techniques such as embryo splitting and nuclear transfer to produce genetically identical individuals. Although embryo splitting is limited to the production of only a few identical individuals, nuclear transfer of donor nuclei into recipient oocytes, whose own nuclear DNA has been removed, can result in large numbers of identical individuals. Moreover, clones can be produced using donor cells from sterile animals, such as steers and geldings, and, unlike their genetic source, these clones are fertile. In reality, due to low efficiencies and the high costs of cloning domestic species, only a limited number of identical individuals are generally produced, and these clones are primarily used as breed stock. In addition to providing a means of rescuing and propagating valuable genetics, somatic cell nuclear transfer (SCNT) research has contributed knowledge that has led to the direct reprogramming of cells (e.g., to induce pluripotent stem cells) and a better understanding of epigenetic regulation during embryonic development. In this review, I provide a broad overview of the historical development of cloning in domestic animals, of its application to the propagation of livestock and transgenic animal production, and of its scientific promise for advancing basic research. PMID:26195770

  4. Artificial cloning of domestic animals

    PubMed Central

    Keefer, Carol L.

    2015-01-01

    Domestic animals can be cloned using techniques such as embryo splitting and nuclear transfer to produce genetically identical individuals. Although embryo splitting is limited to the production of only a few identical individuals, nuclear transfer of donor nuclei into recipient oocytes, whose own nuclear DNA has been removed, can result in large numbers of identical individuals. Moreover, clones can be produced using donor cells from sterile animals, such as steers and geldings, and, unlike their genetic source, these clones are fertile. In reality, due to low efficiencies and the high costs of cloning domestic species, only a limited number of identical individuals are generally produced, and these clones are primarily used as breed stock. In addition to providing a means of rescuing and propagating valuable genetics, somatic cell nuclear transfer (SCNT) research has contributed knowledge that has led to the direct reprogramming of cells (e.g., to induce pluripotent stem cells) and a better understanding of epigenetic regulation during embryonic development. In this review, I provide a broad overview of the historical development of cloning in domestic animals, of its application to the propagation of livestock and transgenic animal production, and of its scientific promise for advancing basic research. PMID:26195770

  5. Cloning goes to the movies.

    PubMed

    Cormick, Craig

    2006-10-01

    Public attitude research conducted by Biotechnology Australia shows that one of the major sources of information on human reproductive cloning is movies. Traditionally, understanding of new and emerging technologies has come through the mass media but human cloning, being so widely addressed through the popular culture of movies, is more effectively defined by Hollywood than the news media or science media. But how well are the science and social issues of cloning portrayed in box office hits such as The Island, Multiplicity, Star Wars: Attack of the Clones and Jurassic Park? These movies have enormous reach and undoubted influence, and are therefore worth analyzing in some detail. This study looks at 33 movies made between 1971 and 2005 that address human reproductive cloning, and it categorizes the films based on their genre and potential influence. Yet rather than simply rating the quality of the science portrayed, the study compares the key messages in these movies with public attitudes towards cloning, to examine the correlations. PMID:17214211

  6. Islamic perspectives on human cloning.

    PubMed

    Sadeghi, Mahmoud

    2007-01-01

    The present paper seeks to assess various views from Islamic jurists relating to human cloning, which is one of the controversial topics in the recent past. Taking Islamic jurisprudence principles, such as the rule of necessity for self preservation and respect for human beings, the rule of la darar wa la dirar ('the necessity to refrain from causing harm to oneself and others') and the rule of usr wa haraj, one may indicate that if human cloning could not be prohibited, as such, it could still be opposed because it gives way to various harmful consequences, which include family disorder, chaos in the clone's family relationships, physical and mental diseases for clones and suffering of egg donors and surrogate mothers. However with due attention to the fact that the reasons behind the prohibition of abortion only restrict the destruction of human embryos in their post-implantation stages, human cloning for biomedical research and exploitation of stem cells from cloned embryos at the blastocyst stage for therapeutic purposes would be acceptable. PMID:17966502

  7. A new dammarane saponin and other triterpenoids from Siolmatra brasiliensis and evaluation of the antidiabetic activity of its extract.

    PubMed

    Dos Santos, Carlos Henrique Corrêa; Borges, Izabeau Pontes; da Silva, Virgínia Claudia; de Sousa, Paulo Teixeira; Kawashita, Nair Honda; Baviera, Amanda Martins; Carvalho, Mario Geraldo de

    2016-09-01

    Context Siolmatra brasiliensis (Cogn.) Baill (Cucurbitaceae) is a climbing plant widely used for the treatment of diabetes mellitus symptoms. Objective This work evaluates the antidiabetic activity of an extract of S. brasiliensis in streptozotocin-diabetic rats and promotes the phytochemical investigation to isolate the major compounds of the same extract. Materials and methods Male Wistar rats were divided into normal (N) and diabetic rats (DC) treated with water; diabetic rats treated with 3U insulin (DI) or with 250 (DSb250) or 500 mg/kg (DSb500) of hydroalcoholic extract of the stalks of S. brasiliensis, via oral gavage, for 21 days. Physiological and biochemical parameters classically altered in diabetes were monitored. The triterpenoids were isolated from the ethyl acetate fraction under silica gel column chromatography and Sephadex-LH20 methods and their structures were determined by NMR, HR-ESI-MS and DC analysis. Results When compared with DC, DSb250 rats showed a reduction in the hyperglycemia (DC: 26.46 ± 0.69 versus DSb250: 19.67 ± 1.06 mmol/L) and glycosuria (DC: 43.02 ± 3.19 versus DSb250: 28.46 ± 2.14 mmol/24 h) and increase in hepatic glycogen (DC: 14.44 ± 1.26 versus DSb250: 22.08 ± 4.26 mg/g). Three known cucurbitacins were isolated from a hydroalcoholic extract of S. brasiliensis, i.e., cayaponosides A1, B4, D, and a new dammarane saponin 3-O-β-d-gentiobiosyl-26-O-β-d-glucopyranosyl-20-hydroxydammar-24-ene. The structures of these compounds were elucidated by spectral data analysis of the natural products and their acetyl derivatives. Discussion and conclusion The known cucurbitacins and/or the new identified saponin may be related with the antidiabetic activity of S. brasiliensis. PMID:26810155

  8. Imperfect Cloning Operations in Algebraic Quantum Theory

    NASA Astrophysics Data System (ADS)

    Kitajima, Yuichiro

    2015-01-01

    No-cloning theorem says that there is no unitary operation that makes perfect clones of non-orthogonal quantum states. The objective of the present paper is to examine whether an imperfect cloning operation exists or not in a C*-algebraic framework. We define a universal -imperfect cloning operation which tolerates a finite loss of fidelity in the cloned state, and show that an individual system's algebra of observables is abelian if and only if there is a universal -imperfect cloning operation in the case where the loss of fidelity is less than . Therefore in this case no universal -imperfect cloning operation is possible in algebraic quantum theory.

  9. Local cloning of entangled states

    SciTech Connect

    Gheorghiu, Vlad; Yu Li; Cohen, Scott M.

    2010-08-15

    We investigate the conditions under which a set S of pure bipartite quantum states on a DxD system can be locally cloned deterministically by separable operations, when at least one of the states is full Schmidt rank. We allow for the possibility of cloning using a resource state that is less than maximally entangled. Our results include that: (i) all states in S must be full Schmidt rank and equally entangled under the G-concurrence measure, and (ii) the set S can be extended to a larger clonable set generated by a finite group G of order |G|=N, the number of states in the larger set. It is then shown that any local cloning apparatus is capable of cloning a number of states that divides D exactly. We provide a complete solution for two central problems in local cloning, giving necessary and sufficient conditions for (i) when a set of maximally entangled states can be locally cloned, valid for all D; and (ii) local cloning of entangled qubit states with nonvanishing entanglement. In both of these cases, we show that a maximally entangled resource is necessary and sufficient, and the states must be related to each other by local unitary 'shift' operations. These shifts are determined by the group structure, so need not be simple cyclic permutations. Assuming this shifted form and partially entangled states, then in D=3 we show that a maximally entangled resource is again necessary and sufficient, while for higher-dimensional systems, we find that the resource state must be strictly more entangled than the states in S. All of our necessary conditions for separable operations are also necessary conditions for local operations and classical communication (LOCC), since the latter is a proper subset of the former. In fact, all our results hold for LOCC, as our sufficient conditions are demonstrated for LOCC, directly.

  10. Local cloning of two product states

    SciTech Connect

    Ji Zhengfeng; Feng Yuan; Ying Mingsheng

    2005-09-15

    Local quantum operations and classical communication (LOCC) put considerable constraints on many quantum information processing tasks such as cloning and discrimination. Surprisingly, however, discrimination of any two pure states survives such constraints in some sense. We show that cloning is not that lucky; namely, probabilistic LOCC cloning of two product states is strictly less efficient than global cloning. We prove our result by giving explicitly the efficiency formula of local cloning of any two product states.

  11. [Cloning and law in Hungary].

    PubMed

    Julesz, Máté

    2015-03-01

    Reproductive human cloning is prohibited in Hungary, as in many other countries. Therapeutic human cloning is not prohibited, just like in many other countries. Stem cell therapy is also allowed. Article III, paragraph (3) of the Hungarian basic law (constitution) strictly forbids total human cloning. Article 1 of the Additional Protocol to the Oviedo Convention, on the Prohibition of Cloning Human Beings (1998) stipulates that any intervention seeking to create a human being genetically identical to another human being, whether living or dead, is prohibited. In Hungary, according to Article 174 of the Criminal Code, total human cloning constitutes a crime. Article 180, paragraph (3) of the Hungarian Act on Health declares that embryos shall not be brought about for research purposes; research shall be conducted only on embryos brought about for reproductive purposes when this is authorized by the persons entitled to decide upon its disposal, or when the embryo is damaged. Article 180, paragraph (5) of the Hungarian Act on Health stipulates that multiple individuals who genetically conform to one another shall not be brought about. According to Article 181, paragraph (1) of the Hungarian Act on Health, an embryo used for research shall be kept alive for not longer than 14 days, not counting the time it was frozen for storage and the time period of research. PMID:25749537

  12. Fat content in migratory central Arizona Brazilian free-tailed bats, Tadarida brasiliensis (Molossidae)

    USGS Publications Warehouse

    O'Shea, Thomas J.

    1976-01-01

    Fat content of migratory Tadarida brasiliensis was determined during the spring, summer and fall of 1972 in the Verde Valley of Arizona. Fat indices were highest in March arrivals, generally declined throughout the summer, and were lowest in September. In both 1972 and 1973 bats had arrived at the study area by mid-March. In 1971 bats were last noted at the area in mid-October while in 1972 they had disappeared by late September. On the basis on physiological calculations it is estimated that bats collected in March 1972 possessed sufficient fat reserves to carry them a mean distance of 716 km north of the study area while September bats had only enough reserves to fly 386 km southward, about 160 km short of the nearest known Sonora wintering locality. It is suggested that in spring the bats may have a more rigidly timed migration and so put on excess fat to counter an uncertain environment to the north. The fall migration may be triggered by more unpredictable events, such as the passage of cold fronts, and less fat reserves may be required for movements into more favorable southern locales.

  13. Antimicrobial activity of benzophenones and extracts from the fruits of Garcinia brasiliensis.

    PubMed

    Naldoni, F J; Claudino, A L R; Cruz, J W; Chavasco, J K; Faria e Silva, P M; Veloso, M P; Dos Santos, M H

    2009-04-01

    The pericarp and seeds from fruits of Garcinia brasiliensis were subjected to extraction with hexane and ethanol. The pericarp hexane extract (PHE) and seed ethanol extract (SEE) were purified by silica gel column chromatography, which permitted isolation of the prenylated benzophenones 7-epiclusianone (1) and guttiferone-A (2), respectively. The antimicrobial activity of PHE, SEE, and compounds 1 and 2 were evaluated against Candida albicans, Staphylococcus aureus, Escherichia coli, and Bacillus cereus cultures. The minimum inhibitory concentration and minimum bactericidal concentration were established. The substances presented activity against S. aureus and B. cereus as follows: PHE, 4.0 microg/mL and 2.4 microg/mL; SEE, 10.0 microg/mL and 12.6 microg/mL; 7-epiclusianone, 1.2 microg/mL and 0.6 microg/mL; and guttiferone-A, 2.4 microg/mL and 2.4 microg/mL, respectively. The direct relationship between the lipophilic character of the structure and activity in Gram-positive bacteria was specifically observed. Therefore these extracts and prenylated benzophenones represent an interesting topic for further studies and open possibilities for an alternative control of diseases associated with Gram-positive bacteria. PMID:19459744

  14. Evolutionary history and identification of conservation units in the giant otter, Pteronura brasiliensis.

    PubMed

    Pickles, R S A; Groombridge, J J; Zambrana Rojas, V D; Van Damme, P; Gottelli, D; Kundu, S; Bodmer, R; Ariani, C V; Iyengar, A; Jordan, W C

    2011-12-01

    The giant otter, Pteronura brasiliensis, occupies a range including the major drainage basins of South America, yet the degree of structure that exists within and among populations inhabiting these drainages is unknown. We sequenced portions of the mitochondrial DNA (mtDNA) cytochrome b (612bp) and control region (383 bp) genes in order to determine patterns of genetic variation within the species. We found high levels of mtDNA haplotype diversity (h = 0.93 overall) and support for subdivision into four distinct groups of populations, representing important centers of genetic diversity and useful units for prioritizing conservation within the giant otter. We tested these results against the predictions of three hypotheses of Amazonian diversification (Pleistocene Refugia, Paleogeography, and Hydrogeology). While the phylogeographic pattern conformed to the predictions of the Refugia Hypothesis, molecular dating using a relaxed clock revealed the phylogroups diverged from one another between 1.69 and 0.84 Ma, ruling out the influence of Late Pleistocene glacial refugia. However, the role of Plio-Pleistocene climate change could not be rejected. While the molecular dating also makes the influence of geological arches according to the Paleogeography Hypothesis extremely unlikely, the recent Pliocene formation of the Fitzcarrald Arch and its effect of subsequently altering drainage pattern could not be rejected. The data presented here support the interactions of both climatic and hydrological changes resulting from geological activity in the Plio-Pleistocene, in shaping the phylogeographic structure of the giant otter. PMID:21925282

  15. Melanins Protect Sporothrix brasiliensis and Sporothrix schenckii from the Antifungal Effects of Terbinafine

    PubMed Central

    Almeida-Paes, Rodrigo; Figueiredo-Carvalho, Maria Helena Galdino; Brito-Santos, Fábio; Almeida-Silva, Fernando; Oliveira, Manoel Marques Evangelista; Zancopé-Oliveira, Rosely Maria

    2016-01-01

    Terbinafine is a recommended therapeutic alternative for patients with sporotrichosis who cannot use itraconazole due to drug interactions or side effects. Melanins are involved in resistance to antifungal drugs and Sporothrix species produce three different types of melanin. Therefore, in this study we evaluated whether Sporothrix melanins impact the efficacy of antifungal drugs. Minimal inhibitory concentrations (MIC) and minimal fungicidal concentrations (MFC) of two Sporothrix brasiliensis and four Sporothrix schenckii strains grown in the presence of the melanin precursors L-DOPA and L-tyrosine were similar to the MIC determined by the CLSI standard protocol for S. schenckii susceptibility to amphotericin B, ketoconazole, itraconazole or terbinafine. When MICs were determined in the presence of inhibitors to three pathways of melanin synthesis, we observed, in four strains, an increase in terbinafine susceptibility in the presence of tricyclazole, a DHN-melanin inhibitor. In addition, one S. schenckii strain grown in the presence of L-DOPA had a higher MFC value when compared to the control. Growth curves in presence of 2×MIC concentrations of terbinafine showed that pyomelanin and, to a lesser extent, eumelanin were able to protect the fungi against the fungicidal effect of this antifungal drug. Our results suggest that melanin protects the major pathogenic species of the Sporothrix complex from the effects of terbinafine and that the development of new antifungal drugs targeting melanin synthesis may improve sporotrichosis therapies. PMID:27031728

  16. In vitro and in vivo photoprotective/photochemopreventive potential of Garcinia brasiliensis epicarp extract.

    PubMed

    Figueiredo, Sônia Aparecida; Vilela, Fernanda Maria Pinto; da Silva, Claudinei Alves; Cunha, Thiago Mattar; Dos Santos, Marcelo Henrique; Fonseca, Maria José Vieira

    2014-02-01

    The damaging effects of sunlight to the skin has triggered studies that involve the synthesis and extraction of organic compounds from natural sources that can absorb UV radiation, and studies on polyphenolic compounds with antioxidant and anti-inflammatory properties that can be used as photochemopreventive agents for reducing skin damage. We investigated the in vitro and in vivo photoprotective/photochemopreventive potential of Garcinia brasiliensis epicarp extract (GbEE). We evaluated the cell viability of L929 fibroblasts after UVB exposure using a quartz plate containing the extract solution or the GbEE formulation. The in vivo photoprotective effect of the GbEE formulation was evaluated by measuring the UVB damage-induced decrease in endogenous reduced glutathione (GSH), the increase in myeloperoxidase (MPO) activity and secretion of cytokines IL-1β and TNF-α. The in vitro methodology using fibroblasts showed that the photoprotective properties of the GbEE solutions and 10% GbEE formulation were similar to the commercial sunscreen (SPF-15). In vivo results demonstrated of the GbEE formulation in decreasing UVB induced-damage such as GSH depletion, an increased in MPO activity and secretion of cytokines IL-1β and TNF-α. The results showed that the extract has great potential for use as a sunscreen in topical formulations in addition to UV filters. PMID:24491421

  17. Aid to a declining matriarch in the giant otter (Pteronura brasiliensis).

    PubMed

    Davenport, Lisa C

    2010-01-01

    Scientists are increasingly revealing the commonalities between the intellectual, emotional and moral capacities of animals and humans. Providing assistance to elderly and ailing family members is a human trait rarely documented for wild animals, other than anecdotal accounts. Here I report observations of multiple forms of assistance to the declining matriarch of a habituated group of giant otters (Pteronura brasiliensis) in Manu National Park, Peru. The otter group had been observed annually for several years and all members were known individually. In 2007, the breeding female of the group failed to reproduce and appeared to be in physical decline. She begged from other family members 43 times over 41 contact hours and received food 11 times. Comparisons with 2004-2006 demonstrate that the family's behavior in 2007 constitutes a role-reversal, in which the majority of assistance and prey transfers accrued from young-to-old rather than from old-to-young. As in human societies, both non-adaptive and adaptive hypotheses could explain the family members' aid to their declining matriarch. I suggest that giant otter families may benefit from the knowledge and experience of an elderly matriarch and "grandparent helper," consistent with the "Grandmother Hypothesis" of adaptive menopause in women. PMID:20613978

  18. Effects of DDE on experimentally poisoned free-tailed bats (Tadarida brasiliensis): Lethal brain concentrations

    USGS Publications Warehouse

    Clark, D.R., Jr.; Kroll, J.C.

    1977-01-01

    Adult female free-tailed bats (Tadarida brasiliensis) were collected at Bracken Cave, Texas, and shipped to the Patuxent Wildlife Research Center. Treated mealworms (Tenebrio molitor) containing 107 ppm DDE were fed to 17 bats; five other bats were fed untreated mealworms. After 40 days on dosage, during which one dosed bat was killed accidentally, four dosed bats were frozen and the remaining 17 were starved to death. The objective was to elevate brain levels of DDE to lethality and measure these concentrations. After the feeding period, dosed bats weighed less than controls. After starvation, the body condition of dosed bats was poorer than that of controls even though there was no difference in the amounts of carcass fat. During starvation, dosed bats lost weight faster than controls. Also, four dosed bats exhibited the prolonged tremoring that characterizes DDE poisoning. DDE increased in brains of starving bats as fat was metabolized. The estimated mean brain concentration of DDE diagnostic of death was 519 ppm with a range of 458-564 ppm. These values resemble diagnostic levels known for two species of passerine birds, but they exceed published levels for two free-tailed bats from Carlsbad Caverns, New Mexico.

  19. Melanins Protect Sporothrix brasiliensis and Sporothrix schenckii from the Antifungal Effects of Terbinafine.

    PubMed

    Almeida-Paes, Rodrigo; Figueiredo-Carvalho, Maria Helena Galdino; Brito-Santos, Fábio; Almeida-Silva, Fernando; Oliveira, Manoel Marques Evangelista; Zancopé-Oliveira, Rosely Maria

    2016-01-01

    Terbinafine is a recommended therapeutic alternative for patients with sporotrichosis who cannot use itraconazole due to drug interactions or side effects. Melanins are involved in resistance to antifungal drugs and Sporothrix species produce three different types of melanin. Therefore, in this study we evaluated whether Sporothrix melanins impact the efficacy of antifungal drugs. Minimal inhibitory concentrations (MIC) and minimal fungicidal concentrations (MFC) of two Sporothrix brasiliensis and four Sporothrix schenckii strains grown in the presence of the melanin precursors L-DOPA and L-tyrosine were similar to the MIC determined by the CLSI standard protocol for S. schenckii susceptibility to amphotericin B, ketoconazole, itraconazole or terbinafine. When MICs were determined in the presence of inhibitors to three pathways of melanin synthesis, we observed, in four strains, an increase in terbinafine susceptibility in the presence of tricyclazole, a DHN-melanin inhibitor. In addition, one S. schenckii strain grown in the presence of L-DOPA had a higher MFC value when compared to the control. Growth curves in presence of 2×MIC concentrations of terbinafine showed that pyomelanin and, to a lesser extent, eumelanin were able to protect the fungi against the fungicidal effect of this antifungal drug. Our results suggest that melanin protects the major pathogenic species of the Sporothrix complex from the effects of terbinafine and that the development of new antifungal drugs targeting melanin synthesis may improve sporotrichosis therapies. PMID:27031728

  20. Aid to a Declining Matriarch in the Giant Otter (Pteronura brasiliensis)

    PubMed Central

    Davenport, Lisa C.

    2010-01-01

    Scientists are increasingly revealing the commonalities between the intellectual, emotional and moral capacities of animals and humans. Providing assistance to elderly and ailing family members is a human trait rarely documented for wild animals, other than anecdotal accounts. Here I report observations of multiple forms of assistance to the declining matriarch of a habituated group of giant otters (Pteronura brasiliensis) in Manu National Park, Peru. The otter group had been observed annually for several years and all members were known individually. In 2007, the breeding female of the group failed to reproduce and appeared to be in physical decline. She begged from other family members 43 times over 41 contact hours and received food 11 times. Comparisons with 2004–2006 demonstrate that the family's behavior in 2007 constitutes a role-reversal, in which the majority of assistance and prey transfers accrued from young-to-old rather than from old-to-young. As in human societies, both non-adaptive and adaptive hypotheses could explain the family members' aid to their declining matriarch. I suggest that giant otter families may benefit from the knowledge and experience of an elderly matriarch and “grandparent helper,” consistent with the “Grandmother Hypothesis” of adaptive menopause in women. PMID:20613978

  1. Biochemical characterization of three phase partitioned naringinase from Aspergillus brasiliensis MTCC 1344.

    PubMed

    Shanmugaprakash, M; Vinothkumar, V; Ragupathy, J; Reddy, D Amala

    2015-09-01

    Naringinase is a complex enzyme composed of α-L-rhamnosidase and β-D-glucosidase, which has a vast potential application in the field of industrial biotechnology. The novel aspect in the present study is employing a three-phase partitioning (TPP) technique for the purification of naringinase by solid-state fermentation using Aspergillus brasiliensis MTCC 1344. At optimum conditions of 28±2 °C and 30% (w/v) ammonium sulfate along with a 1:1 ratio of t-butanol to crude extract, the purification is enhanced by 4.2-fold .Temperature and pH profile of TPP purified naringinase was found to be active with an optimal activity of 719.6 units at an elevated temperature of 60 °C. The kinetic constants K(m) and V(max) using naringin as substrate were 3.21 mM and 321 U/ml. The purified enzyme was not inhibited by any metal ions except Hg(2+) but completely inhibited by adding chelating agents such as EDTA and SDS at a concentration of 10 mM. These results can be inevitable to establish the TPP method to be an inexpensive, economical and attractive technology for better recovery and to find its application in the industrial sector. PMID:26143121

  2. Recombinagenic activity of integerrimine, a pyrrolizidine alkaloid from Senecio brasiliensis, in somatic cells of Drosophila melanogaster.

    PubMed

    Campesato, V R; Graf, U; Reguly, M L; de Andrade, H H

    1997-01-01

    Integerrimine (ITR), a pyrrolizidine alkaloid from Senecio brasiliensis, was tested for genotoxicity using the wing somatic mutation and recombination test (SMART) in Drosophila melanogaster. The compound was administered by chronic feeding (48 hours) of 3-day-old larvae. Two different crosses involving the markers flare (flr) and multiple wing hairs (mwh) were used, that is, the standard (ST) cross and the high bioactivation (HB) cross, which has a high cytochrome P450-dependent bioactivation capacity. In both crosses, the wings of two types of progeny were analyzed, that is, inversion-free marker heterozygotes and balancer heterozygotes carrying multiple inversions. ITR was found to be equally potent in inducing spots in a dose-related manner in the marker heterozygotes of both crosses. This indicates that the bioactivation capacity present in larvae of the ST cross is sufficient to reveal the genotoxic activity of ITR. In the balancer heterozygotes of both crosses, where all recombinational events are eliminated due to the inversions, the frequencies of induced spots were considerably reduced which documents the recombinagenic activity of ITR. Linear regression analysis of the dose response relationships for both genotypes shows that 85% to 90% of the wing spots are due to mitotic recombination. PMID:9020312

  3. Metazoan parasites of Geophagus brasiliensis (Perciformes: Cichlidae) in Patos Lagoon, extreme south of Brazil.

    PubMed

    Rassier, Gabriela Lopes; Pesenti, Tatiana Cheuiche; Pereira Júnior, Joaber; da Silva, Diego Silva; Wendt, Emília Welter; Monteiro, Cassandra de Moraes; Berne, Maria Elizabeth Aires

    2015-01-01

    This study has evaluated the parasitic fauna of 79 pearl cichlids (Geophagus brasiliensis) from the estuary of Patos Lagoon (31° 57' S and 52° 06' W), Rio Grande do Sul, Brazil, during the months of May and June in 2011 and 2012. All the hosts analyzed were infected with at least one species of parasite. A total of eleven metazoa were identified in 459 specimens collected. The trematode Austrodiplostomum compactum (34.2%) and ergasilids Ergasilus lizae (32.9%) and Gauchergasilus lizae (32.9%) were the most prevalent species. The trematodes Thometrema overstreeti and Posthodiplostomum sp. had significantly higher prevalence in fish longer than 20 cm. The sex of the host had no effect on parasite prevalence and abundance. Pearl cichlids are registered as a new host for the trematodes Lobatostoma sp., Homalometron pseudopallidum and Thometrema overstreeti, for the ergasilids Ergasilus lizae and Gauchergasilus euripedesi and for the argulid Argulus spinolosus. The crustacean E. lizae is recorded in Rio Grande do Sul for the first time. PMID:26648010

  4. Analysis of Differentially Expressed Genes Associated with Coronatine-Induced Laticifer Differentiation in the Rubber Tree by Subtractive Hybridization Suppression.

    PubMed

    Zhang, Shi-Xin; Wu, Shao-Hua; Chen, Yue-Yi; Tian, Wei-Min

    2015-01-01

    The secondary laticifer in the secondary phloem is differentiated from the vascular cambia of the rubber tree (Hevea brasiliensis Muell. Arg.). The number of secondary laticifers is closely related to the rubber yield potential of Hevea. Pharmacological data show that jasmonic acid and its precursor linolenic acid are effective in inducing secondary laticifer differentiation in epicormic shoots of the rubber tree. In the present study, an experimental system of coronatine-induced laticifer differentiation was developed to perform SSH identification of genes with differential expression. A total of 528 positive clones were obtained by blue-white screening, of which 248 clones came from the forward SSH library while 280 clones came from the reverse SSH library. Approximately 215 of the 248 clones and 171 of the 280 clones contained cDNA inserts by colony PCR screening. A total of 286 of the 386 ESTs were detected to be differentially expressed by reverse northern blot and sequenced. Approximately 147 unigenes with an average length of 497 bp from the forward and 109 unigenes with an average length of 514 bp from the reverse SSH libraries were assembled and annotated. The unigenes were associated with the stress/defense response, plant hormone signal transduction and structure development. It is suggested that Ca2+ signal transduction and redox seem to be involved in differentiation, while PGA and EIF are associated with the division of cambium initials for COR-induced secondary laticifer differentiation in the rubber tree. PMID:26147807

  5. Quantum cloning machines and the applications

    NASA Astrophysics Data System (ADS)

    Fan, Heng; Wang, Yi-Nan; Jing, Li; Yue, Jie-Dong; Shi, Han-Duo; Zhang, Yong-Liang; Mu, Liang-Zhu

    2014-11-01

    No-cloning theorem is fundamental for quantum mechanics and for quantum information science that states an unknown quantum state cannot be cloned perfectly. However, we can try to clone a quantum state approximately with the optimal fidelity, or instead, we can try to clone it perfectly with the largest probability. Thus various quantum cloning machines have been designed for different quantum information protocols. Specifically, quantum cloning machines can be designed to analyze the security of quantum key distribution protocols such as BB84 protocol, six-state protocol, B92 protocol and their generalizations. Some well-known quantum cloning machines include universal quantum cloning machine, phase-covariant cloning machine, the asymmetric quantum cloning machine and the probabilistic quantum cloning machine. In the past years, much progress has been made in studying quantum cloning machines and their applications and implementations, both theoretically and experimentally. In this review, we will give a complete description of those important developments about quantum cloning and some related topics. On the other hand, this review is self-consistent, and in particular, we try to present some detailed formulations so that further study can be taken based on those results.

  6. Paracoccidioides lutzii Plp43 Is an Active Glucanase with Partial Antigenic Identity with P. brasiliensis gp43

    PubMed Central

    Leitão, Natanael P.; Vallejo, Milene C.; Conceição, Palloma M.; Camargo, Zoilo P.; Hahn, Rosane; Puccia, Rosana

    2014-01-01

    Background Paracoccidioides brasiliensis and P. lutzii cause paracoccidioidomycosis (PCM). P. brasiliensis main diagnostic antigen is glycoprotein gp43, and its peptide sequence is 81% identical with a P. lutzii ortholog here called Plp43. P. lutzii (“Pb01-like”) apparently predominates in Midwestern/Northern Brazil, where high percentages of false-negative reactions using P. brasiliensis antigens have recently been reported. The aim of this work was to produce recombinant Plp43 to study its antigenic identity with gp43. Methodology We expressed rPlp43 as a secreted major component in Pichia pastoris and studied its reactivity in immunoblot with PCM patients' sera from Southwestern and Midwestern Brazil. Principal Findings We showed that rPlp43 is not glycosylated and bears glucanase activity. The protein did not react with anti-gp43 monoclonal antibodies in immunoblot, suggesting absence of the corresponding gp43 epitopes. Nevertheless, common epitope(s) might exist, considering that gp43-positive PCM sera recognized rPlp43 in immunoblot, while gp43-negative sera (33 out of 51) from patients resident in Midwestern Brazil were also rPlp43-negative. Two genotyped P. lutzii were from patients with gp43-negative sera, suggesting that non-reactive sera are from patients infected with this species. Conclusion Our data suggest that gp43 and Plp43 bear one or only a few common epitopes and that gp43 cannot be used in diagnosis of PCM patients infected with P. lutzii probably because Plp43 is poorly expressed during infection. PMID:25166744

  7. Royal Sun Medicinal Mushroom, Agaricus brasiliensis (Agaricomycetidae), Derived Polysaccharides Exert Immunomodulatory Activities In Vitro and In Vivo.

    PubMed

    Fang, Leilei; Zhang, Yanqing; Xie, Junbo; Wang, Lijuan; Zhang, Huan; Wei, Weilu; Li, Yingrui

    2016-01-01

    The royal sun mushroom, Agaricus brasiliensis is a widely consumed mushroom around the world. In this study, the immunoregulatory potential of A. brasiliensis polysaccharides was investigated in vitro and in vivo. In vivo, the polysaccharides remarkably increased the spleen and thymus indexes in mice, and this effect was influenced significantly by age (the adult and the juvenile). The spleen index increased by 27.28% in adult mice treated with the polysaccharides, whereas the increase in juvenile mice was just 12.59% at the dose of 150 mg·kg-1·d-1. Moreover, the effect of the polysaccharides on the thymus and spleen indexes in adult mice was obvious both in males and females. The carbon clearance ability (phagocytic index) was improved with increasing doses, (32.81% at 120 mg·kg-1·d-1, and 38.34% at 150 mg·kg-1·d-1) in mice treated with the polysaccharides. In vitro, the polysaccharides increased the RAW264.7 cell proliferation with 34.78% at 25 µg/mL and 26.78% at 50 µg/mL. Furthermore, the polysaccharides also promoted mRNA expressions of interleukin (IL)-6, IL-1β, cyclooxygenase-2, and Toll-like receptor 4 (TLR4), myeloid differentiation 88 (MYD88), and TIR-domain-containing adapter-inducing interferon-β (TRIF) in the cells, indicating that the polysaccharides induce the secretion of inflammatory cytokines by stimulating TLR4/MyD88 and TLR4/TRIF pathways. In conclusion, these results suggest that A. brasiliensis polysaccharides induce a very promising immunostimulation effect in vivo and in vitro. Therefore, it should be explored as a novel natural functional food additive. PMID:27279534

  8. Spatial analysis of egg distribution and geographic changes in the spawning habitat of the Brazilian sardine Sardinella brasiliensis.

    PubMed

    Gigliotti, E S; Gherardi, D F M; Paes, E T; Souza, R B; Katsuragawa, M

    2010-12-01

    This paper establishes the spawning habitat of the Brazilian sardine Sardinella brasiliensis and investigates the spatial variability of egg density and its relation with oceanographic conditions in the shelf of the south-east Brazil Bight (SBB). The spawning habitats of S. brasiliensis have been defined in terms of spatial models of egg density, temperature-salinity plots, quotient (Q) analysis and remote sensing data. Quotient curves (Q(C)) were constructed using the geographic distribution of egg density, temperature and salinity from samples collected during nine survey cruises between 1976 and 1993. The interannual sea surface temperature (SST) variability was determined using principal component analysis on the SST anomalies (SSTA) estimated from remote sensing data over the period between 1985 and 2007. The spatial pattern of egg occurrences in the SBB indicated that the largest concentration occurred between Paranaguá and São Sebastião. Spawning habitat expanded and contracted during the years, fluctuating around Paranaguá. In January 1978 and January 1993, eggs were found nearly everywhere along the inner shelf of the SBB, while in January 1988 and 1991 spawning had contracted to their southernmost position. The SSTA maps for the spawning periods showed that in the case of habitat expansion (1993 only) anomalies over the SBB were zero or slightly negative, whereas for the contraction period anomalies were all positive. Sardinella brasiliensis is capable of exploring suitable spawning sites provided by the entrainment of the colder and less-saline South Atlantic Central Water onto the shelf by means of both coastal wind-driven (to the north-east of the SBB) and meander-induced (to the south-west of the SBB) upwelling. PMID:21155781

  9. Antifungal activity of extracts of some plants used in Brazilian traditional medicine against the pathogenic fungus Paracoccidioides brasiliensis.

    PubMed

    Johann, Susana; Cisalpino, Patricia Silva; Watanabe, Gisele Almeida; Cota, Betania Barros; de Siqueira, Ezequias Pessoa; Pizzolatti, Moacir Geraldo; Zani, Carlos Leomar; de Resende, Maria Aparecida

    2010-04-01

    Paracoccidioidomycosis (PCM) is a systemic granulomatous disease caused by Paracoccidioides brasiliensis Almeida (Onygenales) that requires 1-2 years of treatment. In the absence of drug therapy, the disease is usually fatal, highlighting the need for the identification of safer, novel, and more effective antifungal compounds. With this need in mind, several plants employed in Brazilian traditional medicine were assayed on P. brasiliensis and murine macrophages. Extracts were prepared from 10 plant species: Inga spp. Mill. (Leguminosae), Schinus terebinthifolius Raddi (Anacardiaceae), Punica granatum L. (Punicaceae), Alternanthera brasiliana Kuntze (Amaranthaceae), Piper regnellii CDC. (Piperaceae), P. abutiloides Kunth (Piperaceae), Herissantia crispa L. Briz. (Malvaceae), Rubus urticaefolius Poir (Rosaceae), Rumex acetosa L. (Polygonaceae), and Baccharis dracunculifolia DC. (Asteraceae). Hexane fractions from hydroalcoholic extracts of Piper regnellii and Baccharis dracunculifolia were the most active against the fungus, displaying minimum inhibitory concentration (MIC) values of 7.8 microg/mL and 7.8-30 mug/mL, respectively. Additionally, neither of the extracts exhibited any apparent cytotoxic effects on murine macrophages at 20 microg/mL. Analyses of these fractions using gas chromatography-mass spectrometry (GC-MS) showed that the major components of B. dracunculifolia were ethyl hydrocinnamate (14.35%) and spathulenol (16.02%), while the major components of the hexane fraction of Piper regnellii were 1-methoxy-4-(1-propenyl) benzene (21.94%) and apiol (21.29%). The activities of these fractions against P. brasiliensis without evidence of cytotoxicity to macrophages justify their investigation as a potential source of new chemical agents for the treatment of PCM. PMID:20645716

  10. Immunization with recombinant Pb27 protein reduces the levels of pulmonary fibrosis caused by the inflammatory response against Paracoccidioides brasiliensis.

    PubMed

    Morais, Elis Araujo; Martins, Estefânia Mara do Nascimento; Boelone, Jankerle Neves; Gomes, Dawidson Assis; Goes, Alfredo Miranda

    2015-02-01

    Paracoccidioidomycosis (PCM) is a systemic mycosis in which the host response to the infectious agent typically consists of a chronic granulomatous inflammatory process. This condition causes lesions that impair lung function and lead to chronic pulmonary insufficiency resulting from fibrosis development, which is a sequel and disabling feature of the disease. The rPb27 protein has been studied for prophylactic and therapeutic treatment against PCM. Previous studies from our laboratory have shown a protective effect of rPb27 against PCM. However, these studies have not determined whether rPb27 immunization prevents lung fibrosis. We therefore conducted this study to investigate fibrosis resulting from infection by Paracoccidioides brasiliensis in the lungs of animals immunized with rPb27. Animals were immunized with rPb27 and subsequently infected with a virulent strain of P. brasiliensis. Fungal load was evaluated by counting colony-forming units, and Masson's trichrome staining was performed to evaluate fibrosis at 30 and 90 days post-infection. The levels of CCR7, active caspase 3, collagen and cytokines were analyzed. At the two time intervals mentioned, the rPb27 group showed lower levels of fibrosis on histology and reduced levels of collagen and the chemokine receptor CCR7 in the lungs. CCR7 was detected at higher levels in the control groups that developed very high levels of pulmonary fibrosis. Additionally, the immunized groups showed high levels of active caspase 3, IFN-γ, TGF-β and IL-10 in the early phase of P. brasiliensis infection. Immunization with Pb27, in addition to its protective effect, was shown to prevent pulmonary fibrosis. PMID:25487973

  11. A study of protein and amino acids in guayule natural rubber

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Natural rubber (NR) from Hevea brasiliensis (the Hevea rubber tree) is a critical agricultural material vital to United States industry, medicine, and defense, yet the country is dependent on NR imports to meet domestic needs. Parthenium argentatum (guayule), a woody desert shrub indigenous to the U...

  12. Development of crops to produce industrially useful natural rubber

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Natural rubber, cis-1,4-polyisoprene, is an essential industrial commodity that most developed countries have to import. Hevea brasiliensis (Hevea), grown in tropical and subtropical areas is the primary source of natural rubber. The high quality and quantity of the rubber cause us to focus on und...

  13. A Six Step Harvesting Procedure of Guayule Small Plots for Laboratory Analyses.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Guayule (Parthenium argentatum Gray) is the best potential source of hypoallergenic latex to replace latex products made with Hevea (Hevea brasiliensis (A. Juss.) Muell.-Arg.) latex that cause Type I allergies. Breeding programs in the past have focused on screening germplasm and developing improved...

  14. Healthy ageing of cloned sheep

    PubMed Central

    Sinclair, K. D.; Corr, S. A.; Gutierrez, C. G.; Fisher, P. A.; Lee, J.-H.; Rathbone, A. J.; Choi, I.; Campbell, K. H. S.; Gardner, D. S.

    2016-01-01

    The health of cloned animals generated by somatic-cell nuclear transfer (SCNT) has been of concern since its inception; however, there are no detailed assessments of late-onset, non-communicable diseases. Here we report that SCNT has no obvious detrimental long-term health effects in a cohort of 13 cloned sheep. We perform musculoskeletal assessments, metabolic tests and blood pressure measurements in 13 aged (7–9 years old) cloned sheep, including four derived from the cell line that gave rise to Dolly. We also perform radiological examinations of all main joints, including the knees, the joint most affected by osteoarthritis in Dolly, and compare all health parameters to groups of 5-and 6-year-old sheep, and published reference ranges. Despite their advanced age, these clones are euglycaemic, insulin sensitive and normotensive. Importantly, we observe no clinical signs of degenerative joint disease apart from mild, or in one case moderate, osteoarthritis in some animals. Our study is the first to assess the long-term health outcomes of SCNT in large animals. PMID:27459299

  15. Clone Poems and the Microcomputer.

    ERIC Educational Resources Information Center

    Irizarry, Estelle

    1989-01-01

    Describes how students can use the computer to study and create clone poems (altering original Spanish-language poems by substituting words and expressions), and how students can gain a deeper appreciation of the original poem's poetic structure and semantics. (CB)

  16. Phase-covariant quantum cloning of qudits

    SciTech Connect

    Fan Heng; Imai, Hiroshi; Matsumoto, Keiji; Wang, Xiang-Bin

    2003-02-01

    We study the phase-covariant quantum cloning machine for qudits, i.e., the input states in a d-level quantum system have complex coefficients with arbitrary phase but constant module. A cloning unitary transformation is proposed. After optimizing the fidelity between input state and single qudit reduced density operator of output state, we obtain the optimal fidelity for 1 to 2 phase-covariant quantum cloning of qudits and the corresponding cloning transformation.

  17. Probabilistic cloning of three symmetric states

    SciTech Connect

    Jimenez, O.; Bergou, J.; Delgado, A.

    2010-12-15

    We study the probabilistic cloning of three symmetric states. These states are defined by a single complex quantity, the inner product among them. We show that three different probabilistic cloning machines are necessary to optimally clone all possible families of three symmetric states. We also show that the optimal cloning probability of generating M copies out of one original can be cast as the quotient between the success probability of unambiguously discriminating one and M copies of symmetric states.

  18. Organochlorine insecticide residues in the free-tailed bat (Tadarida brasiliensis) at Bracken Cave, Texas

    USGS Publications Warehouse

    Clark, D.R., Jr.; Martin, C.O.; Swineford, D.M.

    1975-01-01

    Fifty-nine free-tailed bats (Tadarida brasiliensis mexicana ) were collected at Bracken Cave, Texas, and analyzed for organochlorine insecticides and polychlorinated biphenyls (PCBs). Residues of DDE in the brain were greater in 12 young collected from the floor than in 15 young taken from the ceiling, but food deprivation, not higher residues in the brain, apparently caused young to fall....Among 18 pregnant females, residues of DDE and DDT were highest in yearlings. The first lactation by yearlings caused their residue loads to drop sharply. Thereafter, increasing age was accompanied by increasing residues but amounts generally did not exceed those in yearlings.....Residue levels in embryos were a function both of levels in the female parent and degree of embryonic development. Residues accumulated rapidly in nursing young, and lactating females may excrete from 1.3 to 16.2 (mean = 4.3) micrograms of DDE in milk per day. Maximum individual residue loads may be attained toward the end of nursing, and mobilization of these residues during southward migration may subject Bracken Cave free-tails to maximum lifetime residues in the brain....Comparison of our data with residue data for the free-tail population at Eagle Creek Cave (Arizona) in 1970 produced the following conclusions: ( 1) residues of DDE appeared similar in pregnant females, embryos, lactating females, and fallen young for the two populations; (2) residues of DDT and dieldrin appeared greater in pregnant females at Bracken Cave; (3) DDE and DDT occurred at greater levels in guano samples from Bracken Cave. On this basis, the population decline observed at Eagle Creek Cave between 1963 and 1969 does not appear to be related to the residues observed in the 1970 samples taken from that cave.

  19. Cloning, expression and antiviral activity of IFNγ from the Australian fruit bat, Pteropus alecto.

    PubMed

    Janardhana, Vijaya; Tachedjian, Mary; Crameri, Gary; Cowled, Chris; Wang, Lin-Fa; Baker, Michelle L

    2012-03-01

    Bats are natural reservoir hosts to a variety of viruses, many of which cause morbidity and mortality in other mammals. Currently there is a paucity of information regarding the nature of the immune response to viral infections in bats, partly due to a lack of appropriate bat specific reagents. IFNγ plays a key role in controlling viral replication and coordinating a response for long term control of viral infection. Here we describe the cloning and expression of IFNγ from the Australian flying fox, Pteropus alecto and the generation of mouse monoclonal and chicken egg yolk antibodies specific to bat IFNγ. Our results demonstrate that P. alecto IFNγ is conserved with IFNγ from other species and is induced in bat splenocytes following stimulation with T cell mitogens. P. alecto IFNγ has antiviral activity on Semliki forest virus in cell lines from P. alecto and the microbat, Tadarida brasiliensis. Additionally recombinant bat IFNγ was able to mitigate Hendra virus infection in P. alecto cells. These results provide the first evidence for an antiviral role for bat IFNγin vitro in addition to the application of important immunological reagents for further studies of bat antiviral immunity. PMID:22093696

  20. Economical phase-covariant cloning of qudits

    SciTech Connect

    Buscemi, Francesco; D'Ariano, Giacomo Mauro; Macchiavello, Chiara

    2005-04-01

    We derive the optimal N{yields}M phase-covariant quantum cloning for equatorial states in dimension d with M=kd+N, k integer. The cloning maps are optimal for both global and single-qudit fidelity. The map is achieved by an 'economical' cloning machine, which works without ancilla.

  1. Local cloning of arbitrarily entangled multipartite states

    SciTech Connect

    Kay, Alastair; Ericsson, Marie

    2006-01-15

    We examine the perfect cloning of nonlocal, orthogonal states using only local operations and classical communication. We provide a complete characterisation of the states that can be cloned under these restrictions, and their relation to distinguishability. We also consider the case of catalytic cloning, which we show provides no enhancement to the set of clonable states.

  2. The rubber tree genome shows expansion of gene family associated with rubber biosynthesis

    PubMed Central

    Lau, Nyok-Sean; Makita, Yuko; Kawashima, Mika; Taylor, Todd D.; Kondo, Shinji; Othman, Ahmad Sofiman; Shu-Chien, Alexander Chong; Matsui, Minami

    2016-01-01

    Hevea brasiliensis Muell. Arg, a member of the family Euphorbiaceae, is the sole natural resource exploited for commercial production of high-quality natural rubber. The properties of natural rubber latex are almost irreplaceable by synthetic counterparts for many industrial applications. A paucity of knowledge on the molecular mechanisms of rubber biosynthesis in high yield traits still persists. Here we report the comprehensive genome-wide analysis of the widely planted H. brasiliensis clone, RRIM 600. The genome was assembled based on ~155-fold combined coverage with Illumina and PacBio sequence data and has a total length of 1.55 Gb with 72.5% comprising repetitive DNA sequences. A total of 84,440 high-confidence protein-coding genes were predicted. Comparative genomic analysis revealed strong synteny between H. brasiliensis and other Euphorbiaceae genomes. Our data suggest that H. brasiliensis’s capacity to produce high levels of latex can be attributed to the expansion of rubber biosynthesis-related genes in its genome and the high expression of these genes in latex. Using cap analysis gene expression data, we illustrate the tissue-specific transcription profiles of rubber biosynthesis-related genes, revealing alternative means of transcriptional regulation. Our study adds to the understanding of H. brasiliensis biology and provides valuable genomic resources for future agronomic-related improvement of the rubber tree. PMID:27339202

  3. Therapeutic cloning and tissue engineering.

    PubMed

    Koh, Chester J; Atala, Anthony

    2004-01-01

    A severe shortage of donor organs available for transplantation in the United States leaves patients suffering from diseased and injured organs with few treatment options. Scientists in the field of tissue engineering apply the principles of cell transplantation, material science, and engineering to construct biological substitutes that will restore and maintain normal function in diseased and injured tissues. Therapeutic cloning, where the nucleus from a donor cell is transferred into an enucleated oocyte in order to extract pluripotent embryonic stem cells, offers a potentially limitless source of cells for tissue engineering applications. The present chapter reviews recent advances that have occurred in therapeutic cloning and tissue engineering and describes applications of these new technologies that may offer novel therapies for patients with end-stage organ failure. PMID:15094294

  4. Development of the Sea Star Echinaster (Othilia) brasiliensis, with Inference on the Evolution of Development and Skeletal Plates in Asteroidea.

    PubMed

    Lopes, Elinia Medeiros; Ventura, Carlos Renato Rezende

    2016-02-01

    We describe the development and juvenile morphology of the sea star Echinaster (Othilia) brasiliensis in order to explore evolutionary developmental modes and skeletal homologies. This species produces large, buoyant eggs (0.6 ± 0.03 mm diameter), and has a typical lecithotrophic brachiolaria larva. The planktonic brachiolaria larva is formed 2-4 days after fertilization, when cilia cover the surface. Early juveniles are completely formed by 18 days of age. Initial growth is supported by maternal nutrients while the stomach continues to develop until 60 days after fertilization, when juveniles reach about 0.5 mm of radius length. The madreporite was observed 88 days after fertilization. In the youngest juvenile skeleton of E. (O.) brasiliensis, the madreporite and odontophore are homologous to those of other recent, non-paxillosid asteroids, and follow the Late Madreporic Mode. The emergence of plates related to the ambulacral system follows the Ocular Plate Rule. The development and juvenile skeletal morphology of this species are similar to those of the few other studied species in the genus Echinaster. This study corroborates the notion that the mode of development--including a short-lived lecithotrophic brachiolaria larva--in all Echinaster species shares a similar pattern that may be conserved throughout the evolutionary history of the group. PMID:26896175

  5. Behavioural evidence of male volatile pheromones in the sex-role reversed wolf spiders Allocosa brasiliensis and Allocosa alticeps

    NASA Astrophysics Data System (ADS)

    Aisenberg, Anita; Baruffaldi, Luciana; González, Macarena

    2010-01-01

    The use of chemical signals in a sexual context is widespread in the animal kingdom. Most studies in spiders report the use of female pheromones that attract potential sexual partners. Allocosa brasiliensis and Allocosa alticeps are two burrowing wolf spiders that show sex-role reversal. Females locate male burrows and initiate courtship before males perform any detectable visual or vibratory signal. So, females of these species would be detecting chemical or mechanical cues left by males. Our objective was to explore the potential for male pheromones to play a role in mate detection in A. brasiliensis and A. alticeps. We designed two experiments. In Experiment 1, we tested the occurrence of male contact pheromones by evaluating female courtship when exposed to empty burrows constructed by males or females (control). In Experiment 2, we tested the existence of male volatile pheromones by evaluating female behaviour when exposed to artificial burrows connected to tubes containing males, females or empty tubes (control). Our results suggest the occurrence of male volatile pheromones that trigger female courtship in both Allocosa species. The sex-role reversal postulated for these wolf spiders could be driving the consequent reversal in typical pheromone-emitter and detector roles expected for spiders.

  6. Thioridazine inhibits gene expression control of the cell wall signaling pathway (CWI) in the human pathogenic fungus Paracoccidioides brasiliensis.

    PubMed

    Jabes, Daniela Leite; de Freitas Oliveira, Ana Claudia; Alencar, Valquíria Campos; Menegidio, Fabiano Bezerra; Reno, Débora Liliane Souza; Santos, Daiene Souza; Barbosa, David Aciole; Vilas Boas, Renata Ozelami; de Oliveira Rodrigues Cunha, Rodrigo Luiz; Rodrigues, Tiago; Costa de Oliveira, Regina; Nunes, Luiz R

    2016-06-01

    Paracoccidioides brasiliensis is a thermodimorphic fungus associated with paracoccidioidomycosis (PCM), the most common systemic mycosis in Latin America. PCM treatment involves a long-term chemotherapeutic approach and relapses occur at an alarming frequency. Moreover, the emergence of strains with increased drug-resistance phenotypes puts constant pressure on the necessity to develop new alternatives to treat systemic mycoses. In this work, we show that the phenothiazine (PTZ) derivative thioridazine (TR) inhibits in vitro growth of P. brasiliensis yeasts at micromolar concentrations. We employed microarray hybridization to examine how TR affects gene expression in this fungus, identifying ~1800 genes that were modulated in response to this drug. Dataset evaluation showed that TR inhibits the expression of genes that control the onset of the cell wall integrity (CWI) response, hampering production of all major structural polysaccharides of the fungal cell wall (chitin, α-glucan and β-glucan). Although TR and other PTZs have been shown to display antimicrobial activity by various mechanisms, inhibition of CWI signaling has not yet been reported for these drugs. Thus, TR may provide a novel approach to treat fungal infections by targeting cell wall biogenesis. PMID:26956010

  7. Communication during copulation in the sex-role reversed wolf spider Allocosa brasiliensis: Female shakes for soliciting new ejaculations?

    PubMed

    Garcia Diaz, Virginia; Aisenberg, Anita; Peretti, Alfredo V

    2015-07-01

    Traditional studies on sexual communication have focused on the exchange of signals during courtship. However, communication between the sexes can also occur during or after copulation. Allocosa brasiliensis is a wolf spider that shows a reversal in typical sex roles and of the usual sexual size dimorphism expected for spiders. Females are smaller than males and they are the roving sex that initiates courtship. Occasional previous observations suggested that females performed body shaking behaviors during copulation. Our objective was to analyze if female body shaking is associated with male copulatory behavior in A. brasiliensis, and determine if this female behavior has a communicatory function in this species. For that purpose, we performed fine-scaled analysis of fifteen copulations under laboratory conditions. We video-recorded all the trials and looked for associations between female and male copulatory behaviors. The significant difference between the time before and after female shaking, in favor of the subsequent ejaculation is analyzed. We discuss if shaking could be acting as a signal to accelerate and motivate palpal insertion and ejaculation, and/or inhibiting male cannibalistic tendencies in this species. PMID:25963301

  8. Dentary Morphological Variation in Clevosaurus brasiliensis (Rhynchocephalia, Clevosauridae) from the Upper Triassic of Rio Grande do Sul, Brazil

    PubMed Central

    Romo de Vivar Martínez, Paula Rosario; Bento Soares, Marina

    2015-01-01

    Clevosaurus was a cosmopolitan rhynchocephalian genus, known from the Late Triassic to the Early Jurassic. In South America this genus is represented by C. brasiliensis, an important component of the Linha São Luiz taphocoenosis, on the top of the Norian Santa Maria 2 Sequence of Southern Brazil. The best preserved and most abundant bone elements of C. brasiliensis are dentaries, in which variations of shape and size are observed. The aim of this study is to describe and evaluate the variation, using geometric morphometrics methods. Geometric morphometric analysis of 10 specimens highlights variations in relative size of the dentary. Most of the variation observed for PC1 (83.3%) is likely related to ontogeny, and PC2 (10.0%) is likely related to taphonomic signatures. The development patterns observed, such as the growth of the dentary, consists of differential growth in length between the posterior portion of the dentary, that grows at a higher rate, regarding the anterior portion of the element. This allometric growth is similar to what is observed in other rhynchocephalians and is accompanied by the allometric skull growth, similar to the trend exhibited by clevosaurs. The taphocoenosis is bimodal (juveniles and adults) with a bias towards adult preservation. Some diagenetic influence is reflected in deformed skulls and this is observed in the tangent-plot. Finally, a strong correlation was detected between the taphonomic signatures and the PC2, regarding specially disarticulation and degree of fragmentation. PMID:25793754

  9. Haematological and immunological effects of repeated dose exposure of rats to integerrimine N-oxide from Senecio brasiliensis.

    PubMed

    Elias, Fabiana; Latorre, Andreia O; Pípole, Fernando; Haraguchi, Mitsue; Górniak, Silvana L; Hueza, Isis M

    2011-09-01

    This study is the first in the literature to focus attention on the possible immunotoxic effect of integerrimine N-oxide content in the butanolic residue (BR) of Senecio brasiliensis, a poisonous hepatotoxic plant that contains pyrrolizidine alkaloids (PAs). PAs have been reported as a pasture and food contaminant and as herbal medicine used worldwide and are responsible for poisoning events in livestock and human beings. After the plant extraction, BR extracted from Senecio brasiliensis was found to contain approximately 70% integerrimine N-oxide by elemental and spectral analyses ((1)H and (13)C NMR), which was administered to adult male Wistar Hannover rats at doses of 3, 6 and 9 mg/kg for 28 days. Body weight gain, food consumption, lymphoid organs, neutrophil analysis, humoural immune response, cellular immune response and lymphocyte analysis were evaluated. Our study showed that integerrimine N-oxide could promote an impairment in the body weight gain, interference with blood cell counts and a reducing T cell proliferative activity in rats; however, no differences in the neutrophil activities, lymphocytes phenotyping and humoural and cellular immune responses were observed. It is concluded that doses of integerrimine N-oxide here employed did not produce marked immunotoxic effects. PMID:21722699

  10. Thermoregulation during flight: body temperature and sensible heat transfer in free-ranging Brazilian free-tailed bats (Tadarida brasiliensis).

    PubMed

    Reichard, Jonathan D; Fellows, Spenser R; Frank, Alexander J; Kunz, Thomas H

    2010-01-01

    Bat wings are important for thermoregulation, but their role in heat balance during flight is largely unknown. More than 80% of the energy consumed during flight generates heat as a by-product, and thus it is expected that bat wings should dissipate large amounts of heat to prevent hyperthermia. We measured rectal (T(r)) and surface (T(s)) temperatures of Brazilian free-tailed bats (Tadarida brasiliensis) as they emerged from and returned to their daytime roosts and calculated sensible heat transfer for different body regions (head, body, wings, and tail membrane). Bats' T(r) decreased from 36.8°C during emergence flights to 34.4°C during returns, and T(s) scaled positively with ambient temperature (T(a)). Total radiative heat loss from bats was significantly greater for a radiative sink to the night sky than for a sink with temperature equal to T(a). We found that free-ranging Brazilian free-tailed bats, on average, do not dissipate heat from their wings by convection but instead dissipate radiative heat (L) to the cloudless night sky during flight ([Formula: see text] W). However, within the range of T(a) measured in this study, T. brasiliensis experienced net heat loss between evening emergence and return flights. Regional hypothermia reduces heat loss from wings that are exposed to potentially high convective fluxes. Additional research is needed to establish the role of wings in evaporative cooling during flight in bats. PMID:21034204

  11. Nippostrongylus brasiliensis: Identification of Intelectin-1 and -2 as Stat6-dependent genes expressed in lung and intestine during infection

    PubMed Central

    Voehringer, David; Stanley, Sarah A.; Cox, Jeffery S.; Completo, Gladys C.; Lowary, Todd L.; Locksley, Richard M.

    2009-01-01

    Elimination of the helminth parasite Nippostrongylus brasiliensis from infected mice is mediated by IL-4 or IL-13 and dependent on the IL-4Rα chain and the transcription factor Stat6 in non-hematopoietic cells. However, it is not clear which Stat6-dependent effector molecules mediate worm expulsion. We identified intelectin-1 and -2 as Stat6-dependent genes that are induced during infection. Intelectins can bind galactofuranose, a sugar present only in microorganisms and might therefore serve as microbial pattern element. To analyze whether constitutive expression of intelectin-1 or -2 leads to accelerated pathogen clearance, transgenic mice were generated which express high levels of these genes selectively in the lung. Infection with N. brasiliensis or Mycobacterium tuberculosis did not result in accelerated pathogen clearance in transgenic as compared to wild-type mice. Further, no significant modulation of the immune response in lung or lymph nodes was observed. Thus, under these conditions, intelectins did not enhance pathogen clearance. PMID:17420014

  12. The stress responsive and morphologically regulated hsp90 gene from Paracoccidioides brasiliensis is essential to cell viability

    PubMed Central

    Nicola, André M; Andrade, Rosângela V; Dantas, Alessandra S; Andrade, Patrícia A; Arraes, Fabrício BM; Fernandes, Larissa; Silva-Pereira, Ildinete; Felipe, Maria Sueli S

    2008-01-01

    Background Paracoccidioides brasiliensis is a dimorphic fungus that causes the most prevalent systemic mycosis in Latin America. The response to heat shock is involved in pathogenesis, as this pathogen switches from mycelium to yeast forms in a temperature dependent fashion that is essential to establish infection. HSP90 is a molecular chaperone that helps in the folding and stabilization of selected polypeptides. HSP90 family members have been shown to present important roles in fungi, especially in the pathogenic species, as an immunodominant antigen and also as a potential antifungal therapeutic target. Results In this work, we decided to further study the Pbhsp90 gene, its expression and role in cell viability because it plays important roles in fungal physiology and pathogenesis. Thus, we have sequenced a Pbhsp90 cDNA and shown that this gene is present on the genome as a single copy. We have also confirmed its preferential expression in the yeast phase and its overexpression during dimorphic transition and oxidative stress. Treatment of the yeast with the specific HSP90 inhibitors geldanamycin and radicicol inhibited growth at 2 and 10 μM, respectively. Conclusion The data confirm that the Pbhsp90 gene encodes a morphologically regulated and stress-responsive protein whose function is essential to cell viability of this pathogen. This work also enforces the potential of HSP90 as a target for antifungal therapies, since the use of HSP90 inhibitors is lethal to the P. brasiliensis yeast cells in a dose-responsive manner. PMID:18808717

  13. Cysteine protease of the nematode Nippostrongylus brasiliensis preferentially evokes an IgE/IgG1 antibody response in rats.

    PubMed Central

    Kamata, I; Yamada, M; Uchikawa, R; Matsuda, S; Arizono, N

    1995-01-01

    Some cysteine proteases such as papain and those of mites and schistosomes have potent allergenic properties. To clarify the allergenicity of nematode cysteine proteases, the enzyme was purified from the intestinal nematode Nippostrongylus brasiliensis using cation exchange chromatography and gel filtration chromatography. The purified protease, of 16 kD and pI 8.5, showed maximum enzyme activity at pH 5.5 and substrate preference for Z-Phe-Arg-MCA. The specific inhibitors of cysteine protease leupeptin, iodoacetic acid, and E-64, completely suppressed the activity, indicating that the purified enzyme belongs to the cysteine protease family. Cysteine protease activity was found not only in somatic extract, but also in the excretory-secretory (ES) product of the nematode. When anti-cysteine protease immunoglobulin isotypes were examined in sera from rats infected with N. brasiliensis, a high level of IgG1 and a lower level of IgE antibody were detected. Depletion of IgG antibodies from the sera using protein G affinity columns resulted in a marked increase in reactivity of anti-cysteine protease IgE with the antigen, possibly due to the removal of competing IgG antibodies. In contrast to IgE and IgG1, production of anti-cysteine protease IgG2a was negligible. These results indicate that the nematode cysteine protease preferentially evokes an IgE/IgG1 antibody response. Images Fig. 2 PMID:7554403

  14. Evaluation of the in vitro and in vivo dimorphism of Sporothrix schenckii, Blastomyces dermatitidis, and Paracoccidioides brasiliensis isolates after preservation in mineral oil.

    PubMed

    Lima, Renata Ferretti de; Santos Brito, Marcelly Maria dos; Schäffer, Guido Manoel Vidal; Lima, Osana Cunha de; Borba, Cintia De Moraes

    2004-06-01

    Morphological differentiation has commanded attention for its putative impact on the pathogenesis of invasive fungal infections. We evaluated in vitro and in vivo the dimorphism from mycelial to yeast-phase of Sporothrix schenckii, Blastomyces dermatitidis and Paracoccidioides brasiliensis isolates, two strains for each species, preserved in mineral oil. S. schenckii strains showed typical micromorphology at 25 degrees C but one strain was unable to complete the dimorphic process in vitro. After in vivo passage through mice the strains had the ability to turn into yeast-like cells and to form colonies on brain-heart infusion medium at 36 degrees C. B. dermatitidis strains grew as dirty white to brownish membranous colonies at 25 degrees C and their micromorphology showed thin filaments with single hyaline conidia. At 36 degrees C the colonies did not differ from those grown at 25 degrees C, but produced a transitional micromorphology. P. brasiliensis strains grew as cream-colored cerebriform colonies at 25 degrees C showing a transitional morphology. B. dermatitidis and P. brasiliensis strains did not turn into yeast-like cells in vivo. The present results demonstrate that B. dermatitidis and P. brasiliensis strains were unable to complete the dimorphic process even after in vivo passage, in contrast to the S. schenckii strain. PMID:15284890

  15. Redescription of Ganaspis brasiliensis (Ihering, 1905), new combination, (Hymenoptera: Figitidae) a natural enemy of the invasive Drosophila suzukii (Matsumura, 1931) (Diptera: Drosophilidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The new combination of Ganaspis brasiliensis (Ihering, 1905) is proposed, and the species is redescribed from historical specimens taken in the Neotropical Region as well as more recent specimens reared from Drosophila suzukii Matsumura, 1931 in South Korea. Drosophila suzukii, otherwise known as th...

  16. Influence of the Paracoccidioides brasiliensis 14-3-3 and gp43 proteins on the induction of apoptosis in A549 epithelial cells

    PubMed Central

    da Silva, Julhiany de Fátima; Vicentim, Juliana; de Oliveira, Haroldo Cesar; Marcos, Caroline Maria; Assato, Patricia Akemi; Andreotti, Patrícia Ferrari; da Silva, Juliana Leal Monteiro; Soares, Christiane Pienna; Benard, Gil; Almeida, Ana Marisa Fusco; Mendes-Giannini, Maria José Soares

    2015-01-01

    The fungal strain Paracoccidioides brasiliensis remains viable inside of epithelial cells and can induce apoptosis in this population. However, until now, the molecules that participate in this process remained unknown. Thus, this study evaluated the contribution of two P. brasiliensis molecules, the 14-3-3 and glycoprotein of 43 kDa proteins, which had been previously described as extracellular matrix adhesins and apoptosis inductors in human pneumocytes. Accordingly, epithelial cells were treated with these molecules for different periods of time and the expression of the apoptosis regulating-proteins Bak, Bax, Bcl-2, p53 and caspases were evaluated by terminal deoxynucleotidyl transferase dUTP nick end labelling, flow cytometry and real-time polymerase chain reaction analysis. Our results demonstrated that treatment with these molecules induces apoptosis signalling in pulmonary epithelial cells, showing the same pattern of programmed cell-death as that observed during infection with P. brasiliensis. Thus, we could conclude that P. brasiliensis uses these molecules as virulence factors that participate not only in the fungal adhesion process to host cells, but also in other important cellular mechanisms such as apoptosis. PMID:26038961

  17. [The identification of mouse cloned SFA DNA].

    PubMed

    Yi, Ning; Wu, Weng Qing; Ni, Zu Mei; Shi, Lu Ji

    2002-12-01

    For some basic investigation and the construction of artificial chromosomes, cloned centromeric DNAs identified on a firm ground are required. Thus, in the present work a preliminary screened clone of 13.5 kb DNA, 6# clone, form a mouse centromeric library contructed previously in our library was futher investigated by FISH and PCR. It was found that mouse 6# cloned SFA DNA, as shown by FISH is a fragment of mouse centromeric DNA. Evidence was also observed that 6# cloned SFA DNA consists of mouse minor satellite DNA and other DNA sequences. PMID:15346991

  18. Optimal quantum cloning via spin networks

    SciTech Connect

    Chen Qing; Cheng Jianhua; Wang Kelin; Du Jiangfeng

    2006-09-15

    In this paper we demonstrate that optimal 1{yields}M phase-covariant cloning quantum cloning is available via free dynamical evolution of spin networks. By properly designing the network and the couplings between spins, we show that optimal 1{yields}M phase-covariant cloning can be achieved if the initial state is prepared as a specific symmetric state. Especially, when M is an odd number, the optimal phase-covariant cloning can be achieved without ancillas. Moreover, we demonstrate that the same framework is capable for optimal 1{yields}2 universal cloning.

  19. No-cloning theorem on quantum logics

    SciTech Connect

    Miyadera, Takayuki; Imai, Hideki

    2009-10-15

    This paper discusses the no-cloning theorem in a logicoalgebraic approach. In this approach, an orthoalgebra is considered as a general structure for propositions in a physical theory. We proved that an orthoalgebra admits cloning operation if and only if it is a Boolean algebra. That is, only classical theory admits the cloning of states. If unsharp propositions are to be included in the theory, then a notion of effect algebra is considered. We proved that an atomic Archimedean effect algebra admitting cloning operation is a Boolean algebra. This paper also presents a partial result, indicating a relation between the cloning on effect algebras and hidden variables.

  20. Probabilistic cloning of three nonorthogonal states

    NASA Astrophysics Data System (ADS)

    Zhang, Wen; Rui, Pinshu; Yang, Qun; Zhao, Yan; Zhang, Ziyun

    2015-04-01

    We study the probabilistic cloning of three nonorthogonal states with equal success probabilities. For simplicity, we assume that the three states belong to a special set. Analytical form of the maximal success probability for probabilistic cloning is calculated. With the maximal success probability, we deduce the explicit form of probabilistic quantum cloning machine. In the case of cloning, we get the unambiguous form of the unitary operation. It is demonstrated that the upper bound for probabilistic quantum cloning machine in (Qiu in J Phys A 35:6931, 2002) can be reached only if the three states are equidistant.

  1. Dielectrophoretic separation of mouse melanoma clones

    PubMed Central

    Sabuncu, Ahmet C.; Liu, Jie A.; Beebe, Stephen J.; Beskok, Ali

    2010-01-01

    Dielectrophoresis (DEP) is employed to differentiate clones of mouse melanoma B16F10 cells. Five clones were tested on microelectrodes. At a specific excitation frequency, clone 1 showed a different DEP response than the other four. Growth rate, melanin content, recovery from cryopreservation, and in vitro invasive studies were performed. Clone 1 is shown to have significantly different melanin content and recovery rate from cryopreservation. This paper reports the ability of DEP to differentiate between two malignant cells of the same origin. Different DEP responses of the two clones could be linked to their melanin content. PMID:20697600

  2. No-cloning theorem on quantum logics

    NASA Astrophysics Data System (ADS)

    Miyadera, Takayuki; Imai, Hideki

    2009-10-01

    This paper discusses the no-cloning theorem in a logicoalgebraic approach. In this approach, an orthoalgebra is considered as a general structure for propositions in a physical theory. We proved that an orthoalgebra admits cloning operation if and only if it is a Boolean algebra. That is, only classical theory admits the cloning of states. If unsharp propositions are to be included in the theory, then a notion of effect algebra is considered. We proved that an atomic Archimedean effect algebra admitting cloning operation is a Boolean algebra. This paper also presents a partial result, indicating a relation between the cloning on effect algebras and hidden variables.

  3. Clone DB: an integrated NCBI resource for clone-associated data.

    PubMed

    Schneider, Valerie A; Chen, Hsiu-Chuan; Clausen, Cliff; Meric, Peter A; Zhou, Zhigang; Bouk, Nathan; Husain, Nora; Maglott, Donna R; Church, Deanna M

    2013-01-01

    The National Center for Biotechnology Information (NCBI) Clone DB (http://www.ncbi.nlm.nih.gov/clone/) is an integrated resource providing information about and facilitating access to clones, which serve as valuable research reagents in many fields, including genome sequencing and variation analysis. Clone DB represents an expansion and replacement of the former NCBI Clone Registry and has records for genomic and cell-based libraries and clones representing more than 100 different eukaryotic taxa. Records provide details of library construction, associated sequences, map positions and information about resource distribution. Clone DB is indexed in the NCBI Entrez system and can be queried by fields that include organism, clone name, gene name and sequence identifier. Whenever possible, genomic clones are mapped to reference assemblies and their map positions provided in clone records. Clones mapping to specific genomic regions can also be searched for using the NCBI Clone Finder tool, which accepts queries based on sequence coordinates or features such as gene or transcript names. Clone DB makes reports of library, clone and placement data on its FTP site available for download. With Clone DB, users now have available to them a centralized resource that provides them with the tools they will need to make use of these important research reagents. PMID:23193260

  4. Polysaccharides from Agaricus bisporus and Agaricus brasiliensis show similarities in their structures and their immunomodulatory effects on human monocytic THP-1 cells

    PubMed Central

    2011-01-01

    Background Mushroom polysaccharides have traditionally been used for the prevention and treatment of a multitude of disorders like infectious illnesses, cancers and various autoimmune diseases. Crude mushroom extracts have been tested without detailed chemical analyses of its polysaccharide content. For the present study we decided to chemically determine the carbohydrate composition of semi-purified extracts from 2 closely related and well known basidiomycete species, i.e. Agaricus bisporus and A. brasiliensis and to study their effects on the innate immune system, in particular on the in vitro induction of pro-inflammatory cytokines, using THP-1 cells. Methods Mushroom polysaccharide extracts were prepared by hot water extraction and precipitation with ethanol. Their composition was analyzed by GC-MS and NMR spectroscopy. PMA activated THP-1 cells were treated with the extracts under different conditions and the production of pro-inflammatory cytokines was evaluated by qPCR. Results Semi-purified polysaccharide extracts of A. bisporus and A. brasiliensis (= blazei) were found to contain (1→6),(1→4)-linked α-glucan, (1→6)-linked β-glucan, and mannogalactan. Their proportions were determined by integration of 1H-NMR signs, and were considerably different for the two species. A. brasiliensis showed a higher content of β-glucan, while A. bisporus presented mannogalactan as its main polysaccharide. The extracts induced a comparable increase of transcription of the pro-inflammatory cytokine genes IL-1β and TNF-α as well as of COX-2 in PMA differentiated THP-1 cells. Pro-inflammatory effects of bacterial LPS in this assay could be reduced significantly by the simultaneous addition of A. brasiliensis extract. Conclusions The polysaccharide preparations from the closely related species A. bisporus and A. brasiliensis show major differences in composition: A. bisporus shows high mannogalactan content whereas A. brasiliensis has mostly β-glucan. Semi

  5. Paracoccidioides brasiliensis Vaccine Formulations Based on the gp43-Derived P10 Sequence and the Salmonella enterica FliC Flagellin▿

    PubMed Central

    Braga, Catarina J. M.; Rittner, Glauce M. G.; Muñoz Henao, Julian E.; Teixeira, Aline F.; Massis, Liliana M.; Sbrogio-Almeida, Maria E.; Taborda, Carlos P.; Travassos, Luiz R.; Ferreira, Luís C. S.

    2009-01-01

    Paracoccidioidomycosis (PCM) is a systemic granulomatous disease caused by the dimorphic fungus Paracoccidioides brasiliensis. Anti-PCM vaccine formulations based on the secreted fungal cell wall protein (gp43) or the derived P10 sequence containing a CD4+ T-cell-specific epitope have shown promising results. In the present study, we evaluated new anti-PCM vaccine formulations based on the intranasal administration of P. brasiliensis gp43 or the P10 peptide in combination with the Salmonella enterica FliC flagellin, an innate immunity agonist binding specifically to the Toll-like receptor 5, in a murine model. BALB/c mice immunized with gp43 developed high-specific-serum immunoglobulin G1 responses and enhanced interleukin-4 (IL-4) and IL-10 levels. On the other hand, mice immunized with recombinant purified flagellins genetically fused with P10 at the central hypervariable domain, either flanked or not by two lysine residues, or the synthetic P10 peptide admixed with purified FliC elicited a prevailing Th1-type immune response based on lung cell-secreted type 1 cytokines. Mice immunized with gp43 and FliC and intratracheally challenged with P. brasiliensis yeast cells had increased fungal proliferation and lung tissue damage. In contrast, mice immunized with the chimeric flagellins and particularly those immunized with P10 admixed with FliC reduced P. brasiliensis growth and lung damage. Altogether, these results indicate that S. enterica FliC flagellin modulates the immune response to P. brasiliensis P10 antigen and represents a promising alternative for the generation of anti-PCM vaccines. PMID:19204092

  6. Influence of N-Glycosylation on the Morphogenesis and Growth of Paracoccidioides brasiliensis and on the Biological Activities of Yeast Proteins

    PubMed Central

    Dos Reis Almeida, Fausto Bruno; Carvalho, Fernanda Caroline; Mariano, Vânia Sammartino; Alegre, Ana Claudia Paiva; Silva, Roberto do Nascimento; Hanna, Ebert Seixas; Roque-Barreira, Maria Cristina

    2011-01-01

    The fungus Paracoccidioides brasiliensis is a human pathogen that causes paracoccidioidomycosis, the most prevalent systemic mycosis in Latin America. The cell wall of P. brasiliensis is a network of glycoproteins and polysaccharides, such as chitin, that perform several functions. N-linked glycans are involved in glycoprotein folding, intracellular transport, secretion, and protection from proteolytic degradation. Here, we report the effects of tunicamycin (TM)-mediated inhibition of N-linked glycosylation on P. brasiliensis yeast cells. The underglycosylated yeasts were smaller than their fully glycosylated counterparts and exhibited a drastic reduction of cell budding, reflecting impairment of growth and morphogenesis by TM treatment. The intracellular distribution in TM-treated yeasts of the P. brasiliensis glycoprotein paracoccin was investigated using highly specific antibodies. Paracoccin was observed to accumulate at intracellular locations, far from the yeast wall. Paracoccin derived from TM-treated yeasts retained the ability to bind to laminin despite their underglycosylation. As paracoccin has N-acetyl-β-d-glucosaminidase (NAGase) activity and induces the production of TNF-α and nitric oxide (NO) by macrophages, we compared these properties between glycosylated and underglycosylated yeast proteins. Paracoccin demonstrated lower NAGase activity when underglycosylated, although no difference was detected between the pH and temperature optimums of the two forms. Murine macrophages stimulated with underglycosylated yeast proteins produced significantly lower levels of TNF-α and NO. Taken together, the impaired growth and morphogenesis of tunicamycin-treated yeasts and the decreased biological activities of underglycosylated fungal components suggest that N-glycans play important roles in P. brasiliensis yeast biology. PMID:22216217

  7. A rabies vaccine adjuvanted with saponins from leaves of the soap tree (Quillaja brasiliensis) induces specific immune responses and protects against lethal challenge.

    PubMed

    Yendo, Anna Carolina A; de Costa, Fernanda; Cibulski, Samuel P; Teixeira, Thais F; Colling, Luana C; Mastrogiovanni, Mauricio; Soulé, Silvia; Roehe, Paulo M; Gosmann, Grace; Ferreira, Fernando A; Fett-Neto, Arthur G

    2016-04-29

    Quillaja brasiliensis (Quillajaceae) is a saponin producing species native from southern Brazil and Uruguay. Its saponins are remarkably similar to those of Q. saponaria, which provides most of the saponins used as immunoadjuvants in vaccines. The immunostimulating capacities of aqueous extract (AE) and purified saponin fraction (QB-90) obtained from leaves of Q. brasiliensis were favorably comparable to those of a commercial saponin-based adjuvant preparation (Quil-A) in experimental vaccines against bovine herpesvirus type 1 and 5, poliovirus and bovine viral diarrhea virus in mice model. Herein, the immunogenicity and protection efficacy of rabies vaccines adjuvanted with Q. brasiliensis AE and its saponin fractions were compared with vaccines adjuvanted with either commercial Quil-A or Alum. Mice were vaccinated with one or two doses (on days 0 and 14) of one of the different vaccines and serum levels of total IgG, IgG1 and IgG2a were quantified over time. A challenge experiment with a lethal dose of rabies virus was carried out with the formulations. Viral RNA detection in the brain of mice was performed by qPCR, and RNA copy-numbers were quantified using a standard curve of in vitro transcribed RNA. All Q. brasiliensis saponin-adjuvanted vaccines significantly enhanced levels of specific IgG isotypes when compared with the no adjuvant group (P ≤ 0.05). Overall, one or two doses of saponin-based vaccine were efficient to protect against the lethal rabies exposure. Both AE and saponin fractions from Q. brasiliensis leaves proved potent immunological adjuvants in vaccines against a lethal challenge with a major livestock pathogen, hence confirming their value as competitive or complementary sustainable alternatives to saponins of Q. saponaria. PMID:27032516

  8. Cloning

    MedlinePlus

    ... mammals. These twins are produced when a fertilized egg splits, creating two or more embryos that carry ... of the donor animal's somatic cell into an egg cell, or oocyte, that has had its own ...

  9. Use of the 27-Kilodalton Recombinant Protein from Paracoccidioides brasiliensis in Serodiagnosis of Paracoccidioidomycosis

    PubMed Central

    Ortiz, B. L.; Díez, S.; Urán, M. E.; Rivas, J. M.; Romero, M.; Caicedo, V.; Restrepo, A.; McEwen, J. G.

    1998-01-01

    Paracoccidioidomycosis (PCM) is one of the most important endemic mycoses in Latin America; it is usually diagnosed by observation and/or isolation of the etiologic agent, Paracoccidioides brasiliensis, as well as by a variety of immunological methods. Although the latter are effective, two circumstances, cross-reactions with other mycotic agents and antigen preparation that is marked by extreme variability among lots, hinder proper standardization of the procedures. To circumvent this lack of reproducibility, molecular biology tools were used to produce a recombinant 27-kDa-molecular-mass antigen from this fungus; a sizable quantity of this antigen was obtained through fermentation of Escherichia coli DH5α, which is capable of expressing the fungal protein. The latter was purified by the Prep-Cell System (Bio-Rad); the recovery rate of the pure protein was approximately 6%. A battery of 160 human serum samples, consisting of 64 specimens taken at the time of diagnosis from patients with PCM representing the various clinical forms plus 15 serum specimens each from patients with histoplasmosis and aspergillosis, 10 each from patients with cryptococcosis and tuberculosis, 6 from patients with coccidioidomycosis, and 40 from healthy subjects, were all tested by an indirect enzyme-linked immunosorbent assay with the purified 27-kDa recombinant protein. The latter was used at a concentration of 1.0 μg/well; there were three serum dilutions (1:1,000, 1:2,000, and 1:4,000). The experiment was repeated at least twice. The average sensitivity for both experiments was 73.4%; in comparison with the healthy subjects, the specificity for PCM patients was 87.5% while for patients with other mycoses, it was 58.7%. Important cross-reactions with sera from patients with aspergillosis and histoplasmosis were detected. The positive predictive value of the test was 90.4%. These results indicate that it is possible to employ recombinant antigenic proteins for the immunologic

  10. Genetic structuring among silverside fish (Atherinella brasiliensis) populations from different Brazilian regions

    NASA Astrophysics Data System (ADS)

    da Silva Cortinhas, Maria Cristina; Kersanach, Ralf; Proietti, Maíra; Dumont, Luiz Felipe Cestari; D'Incao, Fernando; Lacerda, Ana Luzia F.; Prata, Pedro Sanmartin; Matoso, Daniele Aparecida; Noleto, Rafael Bueno; Ramsdorf, Wanessa; Boni, Talge Aiex; Prioli, Alberto José; Cestari, Marta Margarete

    2016-09-01

    Estuaries are dynamic environments, key for the survival of innumerous ecologically or economically important fish species. Among these species are Neotropical silversides (Atherinella brasiliensis), which are resident and abundant in Brazilian estuaries and used as a complementary source of income and food for local communities. To better understand silverside populations in Brazil, we evaluated the genetic diversity, structure and demography of fish sampled at six estuaries from the northeastern to the southern coast, using Random Amplified Polymorphic DNA and mitochondrial DNA (D-loop) markers. High haplotype diversities (h ranging from 0.75 to 0.99) were found in all populations except Carapebus, located in Southeast Brazil (h = 0.54). A total of 69 mtDNA haplotypes were found, with Itaparica (Northeast Brazil) and Carapebus presenting only exclusive haplotypes, while some were shared among populations in the South. Strong regional structure was observed, with very high differentiation between Itaparica and Carapebus, as well as among these two populations and the ones from the Southern region (Paranaguá, Conceição, Camacho and Patos). Among southern areas, low/moderate structure was detected. Most populations showed unimodal mismatch distributions indicating recent demographic expansion, while Carapebus presented a multimodal distribution characteristic of a stable or bottlenecked population. Times since possible population expansion were highest in Itaparica (32,500 ya) and Carapebus (29,540 ya), while in the Southern region longest time was observed at Conceição (25,540 ya) and shortest at Patos (9720 ya). In a general manner, haplotype diversities were directly related to times since population expansions; again, Carapebus was the exception, displaying long time since expansion but low diversity, possibly due to a recent bottleneck caused by the isolation and human impacts this lagoon is subject to. Isolation by Distance was significant for Itaparica

  11. The cochlea of Tadarida brasiliensis: specialized functional organization in a generalized bat.

    PubMed

    Vater, M; Siefer, W

    1995-11-01

    Tadarida brasiliensis mexicana employs a broad-band sonar system at frequencies between 80 and 20 kHz and is characterized by non-specialized hearing capabilities. The cochlear frequency map was determined with extracellular horseradish peroxidase tracing in relation to quantitative morphological data obtained with light, scanning and transmission electron microscopy. These data reveal distinct species characteristic specializations clearly separate from the patterns observed in other bats with either broad-band or narrow-band sonar systems. The basilar membrane (BM) is coiled to 2.5 turns and about 12 mm long. Its thickness and width only change within the extreme basal and apical ends. The frequency range from about 30 to 80 kHz is represented in the lower basal turn with a typically mammalian mapping coefficient of about 3 mm/octave. This region exhibits morphological features correlated with non-specialized processing of high frequencies. (1) The BM is radially segmented by thickenings of pars tecta and pars pectinata. (2) The 3 rows of outer hair cells (OHCs) have similar morphology. Between 35 and 86% distance from base, frequencies between 30 and 12 kHz are represented with a slightly expanded mapping coefficient of about 6 mm/octave. In analogy to previous work, this cochlea region is termed acoustic fovea. It includes the frequency range of maximum sensitivity and sharpest tuning (21-27 kHz) but also frequencies below the sonar signals. The fovea is characterized by several morphological specializations. (1) The BM features a continuous radial thickening mainly composed of hyaline substance. (2) There is an increased number of layers of tension fibroblasts in the spiral ligament. (3) There are morphological differences in the arrangements of stereocilia bundles among the 3 rows of OHCs. The transitions between non-specialized and specialized cochlear regions occur gradually within a distance of about 600 microns. The gradients in stereocilia length of both

  12. Hybrid Sequencing Approach Applied to Human Fecal Metagenomic Clone Libraries Revealed Clones with Potential Biotechnological Applications

    PubMed Central

    Džunková, Mária; D’Auria, Giuseppe; Pérez-Villarroya, David; Moya, Andrés

    2012-01-01

    Natural environments represent an incredible source of microbial genetic diversity. Discovery of novel biomolecules involves biotechnological methods that often require the design and implementation of biochemical assays to screen clone libraries. However, when an assay is applied to thousands of clones, one may eventually end up with very few positive clones which, in most of the cases, have to be “domesticated” for downstream characterization and application, and this makes screening both laborious and expensive. The negative clones, which are not considered by the selected assay, may also have biotechnological potential; however, unfortunately they would remain unexplored. Knowledge of the clone sequences provides important clues about potential biotechnological application of the clones in the library; however, the sequencing of clones one-by-one would be very time-consuming and expensive. In this study, we characterized the first metagenomic clone library from the feces of a healthy human volunteer, using a method based on 454 pyrosequencing coupled with a clone-by-clone Sanger end-sequencing. Instead of whole individual clone sequencing, we sequenced 358 clones in a pool. The medium-large insert (7–15 kb) cloning strategy allowed us to assemble these clones correctly, and to assign the clone ends to maintain the link between the position of a living clone in the library and the annotated contig from the 454 assembly. Finally, we found several open reading frames (ORFs) with previously described potential medical application. The proposed approach allows planning ad-hoc biochemical assays for the clones of interest, and the appropriate sub-cloning strategy for gene expression in suitable vectors/hosts. PMID:23082187

  13. Detectability of Plasmodium falciparum clones

    PubMed Central

    2010-01-01

    Background In areas of high transmission people often harbour multiple clones of Plasmodium falciparum, but even PCR-based diagnostic methods can only detect a fraction (the detectability, q) of all clones present in a host. Accurate measurements of detectability are desirable since it affects estimates of multiplicity of infection, prevalence, and frequency of breakthrough infections in clinical drug trials. Detectability can be estimated by typing repeated samples from the same host but it has been unclear what should be the time interval between the samples and how the data should be analysed. Methods A longitudinal molecular study was conducted in the Kassena-Nankana district in northern Ghana. From each of the 80 participants, four finger prick samples were collected over a period of 8 days, and tested for presence of different Merozoite Surface Protein (msp) 2 genotypes. Implications for estimating q were derived from these data by comparing the fit of statistical models of serial dependence and over-dispersion. Results The distribution of the frequencies of detection for msp2 genotypes was close to binomial if the time span between consecutive blood samples was at least 7 days. For shorter intervals the probabilities of detection were positively correlated, i.e. the shorter the interval between two blood collections, the more likely the diagnostic results matched for a particular genotype. Estimates of q were rather insensitive to the statistical model fitted. Conclusions A simple algorithm based on analysing blood samples collected 7 days apart is justified for generating robust estimates of detectability. The finding of positive correlation of detection probabilities for short time intervals argues against imperfect detection being directly linked to the 48-hour periodicity of P. falciparum. The results suggest that the detectability of a given parasite clone changes over time, at an unknown rate, but fast enough to regard blood samples taken one week

  14. Δ24-Sterol Methyltransferase Plays an Important Role in the Growth and Development of Sporothrix schenckii and Sporothrix brasiliensis

    PubMed Central

    Borba-Santos, Luana P.; Visbal, Gonzalo; Gagini, Thalita; Rodrigues, Anderson M.; de Camargo, Zoilo P.; Lopes-Bezerra, Leila M.; Ishida, Kelly; de Souza, Wanderley; Rozental, Sonia

    2016-01-01

    Inhibition of Δ24-sterol methyltransferase (24-SMT) in Sporothrix schenckii sensu stricto and Sporothrix brasiliensis was investigated in vitro. The effects on fungal growth and sterol composition of the 24-SMT inhibitor 22-hydrazone-imidazolin-2-yl-chol-5-ene-3β-ol (H3) were compared to those of itraconazole. MIC and MFC analysis showed that H3 was more effective than itraconazole against both species in both their filamentous and yeast forms. H3 showed fungistatic activity in a time-kill assay, with inhibitory activity stronger than that of itraconazole. GC analysis of cell sterol composition showed that sterols present in control cells (ergosterol and precursors) were completely replaced by 14α-methylated sterols after H3 exposure. Itraconazole only partially inhibited ergosterol synthesis but completely arrested synthesis of other sterols found in control cells, promoting accumulation of nine 14α-methyl sterols. Based on these results, we propose a schematic model of sterol biosynthesis pathways in S. schenckii and S. brasiliensis. Effects on cell morphology due to 24-SMT inhibition by H3 as analyzed by SEM and TEM included irregular cell shape, reduced cytoplasmic electron-density, and reduced thickness of the microfibrillar cell wall layer. Moreover, 24-SMT inhibition by H3 promoted mitochondrial disturbance, as demonstrated by alterations in MitoTracker® Red CMXRos fluorescence intensity evaluated by flow cytometry. When used in conjunction with itraconazole, H3 enhanced the effectiveness of itraconazole against all tested strains, reducing at least half (or more) the MIC values of itraconazole. In addition, cytotoxicity assays revealed that H3 was more selective toward these fungi than was itraconazole. Thus, 24-SMT inhibition by H3 was an effective antifungal strategy against S. schenckii and S. brasiliensis. Inhibition of the methylation reaction catalyzed by 24-SMT has a strong antiproliferative effect via disruption of ergosterol homeostasis

  15. Δ(24)-Sterol Methyltransferase Plays an Important Role in the Growth and Development of Sporothrix schenckii and Sporothrix brasiliensis.

    PubMed

    Borba-Santos, Luana P; Visbal, Gonzalo; Gagini, Thalita; Rodrigues, Anderson M; de Camargo, Zoilo P; Lopes-Bezerra, Leila M; Ishida, Kelly; de Souza, Wanderley; Rozental, Sonia

    2016-01-01

    Inhibition of Δ(24)-sterol methyltransferase (24-SMT) in Sporothrix schenckii sensu stricto and Sporothrix brasiliensis was investigated in vitro. The effects on fungal growth and sterol composition of the 24-SMT inhibitor 22-hydrazone-imidazolin-2-yl-chol-5-ene-3β-ol (H3) were compared to those of itraconazole. MIC and MFC analysis showed that H3 was more effective than itraconazole against both species in both their filamentous and yeast forms. H3 showed fungistatic activity in a time-kill assay, with inhibitory activity stronger than that of itraconazole. GC analysis of cell sterol composition showed that sterols present in control cells (ergosterol and precursors) were completely replaced by 14α-methylated sterols after H3 exposure. Itraconazole only partially inhibited ergosterol synthesis but completely arrested synthesis of other sterols found in control cells, promoting accumulation of nine 14α-methyl sterols. Based on these results, we propose a schematic model of sterol biosynthesis pathways in S. schenckii and S. brasiliensis. Effects on cell morphology due to 24-SMT inhibition by H3 as analyzed by SEM and TEM included irregular cell shape, reduced cytoplasmic electron-density, and reduced thickness of the microfibrillar cell wall layer. Moreover, 24-SMT inhibition by H3 promoted mitochondrial disturbance, as demonstrated by alterations in MitoTracker(®) Red CMXRos fluorescence intensity evaluated by flow cytometry. When used in conjunction with itraconazole, H3 enhanced the effectiveness of itraconazole against all tested strains, reducing at least half (or more) the MIC values of itraconazole. In addition, cytotoxicity assays revealed that H3 was more selective toward these fungi than was itraconazole. Thus, 24-SMT inhibition by H3 was an effective antifungal strategy against S. schenckii and S. brasiliensis. Inhibition of the methylation reaction catalyzed by 24-SMT has a strong antiproliferative effect via disruption of ergosterol homeostasis

  16. Unified universal quantum cloning machine and fidelities

    SciTech Connect

    Wang Yinan; Shi Handuo; Xiong Zhaoxi; Jing Li; Mu Liangzhu; Ren Xijun; Fan Heng

    2011-09-15

    We present a unified universal quantum cloning machine, which combines several different existing universal cloning machines together, including the asymmetric case. In this unified framework, the identical pure states are projected equally into each copy initially constituted by input and one half of the maximally entangled states. We show explicitly that the output states of those universal cloning machines are the same. One importance of this unified cloning machine is that the cloning procession is always the symmetric projection, which reduces dramatically the difficulties for implementation. Also, it is found that this unified cloning machine can be directly modified to the general asymmetric case. Besides the global fidelity and the single-copy fidelity, we also present all possible arbitrary-copy fidelities.

  17. Quantum cloning disturbed by thermal Davies environment

    NASA Astrophysics Data System (ADS)

    Dajka, Jerzy; Łuczka, Jerzy

    2016-03-01

    A network of quantum gates designed to implement universal quantum cloning machine is studied. We analyze how thermal environment coupled to auxiliary qubits, `blank paper' and `toner' required at the preparation stage of copying, modifies an output fidelity of the cloner. Thermal environment is described in terms of the Markovian Davies theory. We show that such a cloning machine is not universal any more but its output is independent of at least a part of parameters of the environment. As a case study, we consider cloning of states in a six-state cryptography's protocol. We also briefly discuss cloning of arbitrary input states.

  18. Quantum cloning disturbed by thermal Davies environment

    NASA Astrophysics Data System (ADS)

    Dajka, Jerzy; Łuczka, Jerzy

    2016-06-01

    A network of quantum gates designed to implement universal quantum cloning machine is studied. We analyze how thermal environment coupled to auxiliary qubits, `blank paper' and `toner' required at the preparation stage of copying, modifies an output fidelity of the cloner. Thermal environment is described in terms of the Markovian Davies theory. We show that such a cloning machine is not universal any more but its output is independent of at least a part of parameters of the environment. As a case study, we consider cloning of states in a six-state cryptography's protocol. We also briefly discuss cloning of arbitrary input states.

  19. Vascular contrast enhanced micro-CT imaging of "radiators" in the Brazilian free-tailed bat (Tadarida brasiliensis).

    PubMed

    Reichard, Jonathan D; Kunz, Thomas H; Keller, Charles; Prajapati, Suresh I

    2012-04-01

    The Brazilian free-tailed bat (Tadarida brasiliensis) exhibits a highly vascularized, hairless thermal window (or "radiator") on the proximal ventral surfaces of extended wings and body. We identified this character using thermal infrared imaging and investigated the vasculature using barium sulfate enhanced microcomputed tomography (micro-CT). Micro-CT images revealed unique arrangements of arteries and veins in the region of the radiator positioned perpendicular to the axis of the body. Coupling micro-CT imaging with analysis of surface temperature profiles, we concluded that radiators aid in thermoregulation during flight in variable environments. This study represents the first application of contrast enhanced micro-CT to visualize vasculature of bats and thus exhibits a promising technique for further investigations of cardiovascular function and anatomy in bats. PMID:22282439

  20. [Embrionary-larval development of the tropical fish Hemirhamphus brasiliensis (Beloniformes: Hemirhamphidae) from eggs collected in the wild].

    PubMed

    Rosas, Jesús; Mata, Ernesto; Velásquez, Aidé; Cabrera, Tomas

    2008-09-01

    The embryo formation and larval development of Hemirhamphus brasiliensis Linnaeus, 1758 (Pisces: Hemirhamphidae) is described from morula stage eggs collected on Sargassum sp. Thalii in the field (10 degrees 50'55.2" N y 64 degrees 09'467" W). The eggs were spherical, 1 923.54 +/- 72.35 microm diameter with several corionic filaments, and are striated. During the first 48 h the embryo developed cephalic vesicle, miomers, and a heart located on the external body surface, beating strongly and circulating colorless blood which became pigmented red later. Before hatching, the larva developed kidney, gut tract, liver and biliar vesicle, pectoral fins, four pairs of gill arches and the mouth. The larva hatched at 114 h, the body was torpedo-shaped, yellow-green, with several dendriform melanophores; the pelvic fin was observed 72 h post hatching. At 240 hours the metamorphoses was completed. When the larvae hatched they could ingest Artemia metanauplii. PMID:19419056

  1. Identification of oversulphated galactosaminoglycans in intestinal-mucosal mast cells of rats infected with the nematode worm Nippostrongylus brasiliensis.

    PubMed Central

    Kusche, M; Lindahl, U; Enerbäck, L; Rodén, L

    1988-01-01

    The oversulphated galactosaminoglycans synthesized by rat mucosal mast cells were isolated from the small intestine of animals infected with the nematode Nippostrongylus brasiliensis, which causes proliferation of these cells. The 35S-labelled polysaccharides were degraded by digestion with chondroitinase ABC, and the structures of the disaccharide products were determined by cleavage with mercuric acetate followed by electrophoretic characterization of the resultant sulphated monosaccharides. It was concluded that about half of the disulphated disaccharide units in the polysaccharide consisted of chondroitin sulphate E-type structures [GlcA-GalNAc(4,6-di-OSO3)], in which both sulphate groups were located on the N-acetylgalactosamine unit. The remainder consisted of isomeric structures with one sulphate group on the N-acetylgalactosamine residue and one on the hexuronic acid unit and presumably represented the dermatan sulphate-type sequence [IdoA(2-OSO3)-GalNAc(4-OSO3)]. PMID:3178741

  2. Biochemical Characterization of Paracoccidioides brasiliensis α-1,3-Glucanase Agn1p, and Its Functionality by Heterologous Expression in Schizosaccharomyces pombe

    PubMed Central

    Villalobos-Duno, Héctor; San-Blas, Gioconda; Paulinkevicius, Maryan; Sánchez-Martín, Yolanda; Nino-Vega, Gustavo

    2013-01-01

    α-1,3-Glucan is present as the outermost layer of the cell wall in the pathogenic yeastlike (Y) form of Paracoccidioides brasiliensis. Based on experimental evidence, this polysaccharide has been proposed as a fungal virulence factor. To degrade α-1,3-glucan and allow remodeling of the cell wall, α-1,3-glucanase is required. Therefore, the study of this enzyme, its encoding gene, and regulatory mechanisms, might be of interest to understand the morphogenesis and virulence process in this fungus. A single gene, orthologous to other fungal α-1,3-glucanase genes, was identified in the Paracoccidioides genome, and labeled AGN1. Transcriptional levels of AGN1 and AGS1 (α-1,3-glucan synthase-encoding gene) increased sharply when the pathogenic Y phase was cultured in the presence of 5% horse serum, a reported booster for cell wall α-1,3-glucan synthesis in this fungus. To study the biochemical properties of P. brasiliensis Agn1p, the enzyme was heterologously overexpressed, purified, and its activity profile determined by means of the degradation of carboxymethyl α-1,3-glucan (SCMG, chemically modified from P. brasiliensis α-1,3-glucan), used as a soluble substrate for the enzymatic reaction. Inhibition assays, thin layer chromatography and enzymatic reactions with alternative substrates (dextran, starch, chitin, laminarin and cellulose), showed that Agn1p displays an endolytic cut pattern and high specificity for SCMG. Complementation of a Schizosaccharomyces pombe agn1Δ strain with the P. brasiliensis AGN1 gene restored the wild type phenotype, indicating functionality of the gene, suggesting a possible role of Agn1p in the remodeling of P. brasiliensis Y phase cell wall. Based on amino acid sequence, P. brasiliensis Agn1p, groups within the family 71 of fungal glycoside hydrolases (GH-71), showing similar biochemical characteristics to other members of this family. Also based on amino acid sequence alignments, we propose a subdivision of fungal GH-71 into at

  3. The transcriptome analysis of early morphogenesis in Paracoccidioides brasiliensis mycelium reveals novel and induced genes potentially associated to the dimorphic process

    PubMed Central

    Bastos, Karinne P; Bailão, Alexandre M; Borges, Clayton L; Faria, Fabricia P; Felipe, Maria SS; Silva, Mirelle G; Martins, Wellington S; Fiúza, Rogério B; Pereira, Maristela; Soares, Célia MA

    2007-01-01

    Background Paracoccidioides brasiliensis is a human pathogen with a broad distribution in Latin America. The fungus is thermally dimorphic with two distinct forms corresponding to completely different lifestyles. Upon elevation of the temperature to that of the mammalian body, the fungus adopts a yeast-like form that is exclusively associated with its pathogenic lifestyle. We describe expressed sequence tags (ESTs) analysis to assess the expression profile of the mycelium to yeast transition. To identify P. brasiliensis differentially expressed sequences during conversion we performed a large-scale comparative analysis between P. brasiliensis ESTs identified in the transition transcriptome and databases. Results Our analysis was based on 1107 ESTs from a transition cDNA library of P. brasiliensis. A total of 639 consensus sequences were assembled. Genes of primary metabolism, energy, protein synthesis and fate, cellular transport, biogenesis of cellular components were represented in the transition cDNA library. A considerable number of genes (7.51%) had not been previously reported for P. brasiliensis in public databases. Gene expression analysis using in silico EST subtraction revealed that numerous genes were more expressed during the transition phase when compared to the mycelial ESTs [1]. Classes of differentially expressed sequences were selected for further analysis including: genes related to the synthesis/remodeling of the cell wall/membrane. Thirty four genes from this family were induced. Ten genes related to signal transduction were increased. Twelve genes encoding putative virulence factors manifested increased expression. The in silico approach was validated by northern blot and semi-quantitative RT-PCR. Conclusion The developmental program of P. brasiliensis is characterized by significant differential positive modulation of the cell wall/membrane related transcripts, and signal transduction proteins, suggesting the related processes important

  4. Purification, characterization, and antioxidant activities of selenium-containing proteins and polysaccharides in royal sun mushroom, Agaricus brasiliensis (Higher Basidiomycetes).

    PubMed

    Mao, Guanghua; Feng, Weiwei; Xiao, Hui; Zhao, Ting; Li, Fang; Zou, Ye; Ren, Yuena; Zhu, Yang; Yang, Liuqing; Wu, Xiangyang

    2014-01-01

    The Agaricus brasiliensis proves to be the main source of many minerals, especially selenium (Se). In this study, Se-containing polysaccharides and proteins were isolated, purified, and characterized. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl radical scavenging activity of Se-containing proteins and polysaccharides were also studied. Selenium in A. brasiliensis is present mostly in organic forms, accounting for 81.57% of the total Se. The organic forms of selenium mainly present in Se proteins account for 73.53%, while 12.23% is in Se polysaccharides. Two Se-containing proteins (AB-SePA-22) and (AB-SePG-22) with Se contents of 4.935 µg/g and 6.083 µg/g were obtained. AB-SePA-22 appeared as four bands with molecular masses of 16.7, 21.7, 26.3, and 33.6 kDa, respectively. The Se content of the three Se-containing polysaccharides, namely AB-SeP-1, AB-SeP-2, and AB-SeP-3, were 1.911, 0.613, and 0.671 µg/g, respectively. AB-SeP-1 (3.1×103 Da) was composed of glucose and galactose in a 7.494:1 molar ratio, whereas AB-SeP-2 (2.1×104 Da and 3.5×104 Da) and AB-SeP-3 (1.1×105 Da) were composed of glucose, galactose, and mannose with molar ratios of 27.01:1.55:1 and 9.805:1:1.22, respectively. Moreover, crude Se polysaccharide and total soluble Se protein had good antioxidant activities on scavenging DPPH and hydroxyl radical, and further research is needed. PMID:25271981

  5. STRU-cloning: a fast, inexpensive and efficient cloning procedure applicable to both small scale and structural genomics size cloning.

    PubMed

    Bellini, Dom; Fordham-Skelton, Anthony P; Papiz, Miroslav Z

    2011-05-01

    We have developed a Single-Tube Restriction-based Ultrafiltration (STRU) cloning procedure that updates traditional ligation-dependent cloning to challenge the newer, faster and more efficient ligation-free techniques and could make it the method of choice. STRU-cloning employs centrifugal filter units with membrane of suitable cut off to remove small unwanted DNA fragments created during restriction of plasmids or PCR products. Heat inactivation, of restriction enzymes, followed by DNA ligation is then performed on the filtrate. By removing the agarose gel electrophoresis DNA purification step from the traditional protocol, which is time consuming and is known to be the cause of ligation problems, STRU-cloning becomes fast, very efficient, inexpensive and offers the highest degree of cloning flexibility by using restriction sites and can be performed in a single tube. This novel agarose gel-free cloning procedure provides benefits for both small and large scale cloning projects. Unlike traditional cloning it can be easily implemented as a fully automated process at very low costs. PMID:21052867

  6. Positional Cloning by Linkage Disequilibrium

    PubMed Central

    Maniatis, Nikolas; Collins, Andrew; Gibson, Jane; Zhang, Weihua; Tapper, William; Morton, Newton E.

    2004-01-01

    Recently, metric linkage disequilibrium (LD) maps that assign an LD unit (LDU) location for each marker have been developed (Maniatis et al. 2002). Here we present a multiple pairwise method for positional cloning by LD within a composite likelihood framework and investigate the operating characteristics of maps in physical units (kb) and LDU for two bodies of data (Daly et al. 2001; Jeffreys et al. 2001) on which current ideas of blocks are based. False-negative indications of a disease locus (type II error) were examined by selecting one single-nucleotide polymorphism (SNP) at a time as causal and taking its allelic count (0, 1, or 2, for the three genotypes) as a pseudophenotype, Y. By use of regression and correlation, association between every pseudophenotype and the allelic count of each SNP locus (X) was based on an adaptation of the Malecot model, which includes a parameter for location of the putative gene. By expressing locations in kb or LDU, greater power for localization was observed when the LDU map was fitted. The efficiency of the kb map, relative to the LDU map, to describe LD varied from a maximum of 0.87 to a minimum of 0.36, with a mean of 0.62. False-positive indications of a disease locus (type I error) were examined by simulating an unlinked causal SNP and the allele count was used as a pseudophenotype. The type I error was in good agreement with Wald’s likelihood theorem for both metrics and all models that were tested. Unlike tests that select only the most significant marker, haplotype, or haploset, these methods are robust to large numbers of markers in a candidate region. Contrary to predictions from tagging SNPs that retain haplotype diversity, the sample with smaller size but greater SNP density gave less error. The locations of causal SNPs were estimated with the same precision in blocks and steps, suggesting that block definition may be less useful than anticipated for mapping a causal SNP. These results provide a guide to

  7. Cloning genes for non-syndromal hearing impairment.

    PubMed

    Smith, R J; Van Camp, G

    1999-10-01

    Over 45 genes that cause autosomal non-syndromic hearing impairment (NSHI) have been localized and many more are predicted to exist. To clone these genes, a number of different strategies can be used. This paper focuses on four general approaches: functional cloning, positional cloning, position-dependent candidate gene cloning, and position-independent candidate gene cloning. PMID:10890140

  8. Reversibility of continuous-variable quantum cloning

    SciTech Connect

    Filip, Radim; Marek, Petr; Fiurasek, Jaromir

    2004-01-01

    We analyze a reversibility of optimal Gaussian 1{yields}2 quantum cloning of a coherent state using only local operations on the clones and classical communication between them and propose a feasible experimental test of this feature. Performing Bell-type homodyne measurement on one clone and anticlone, an arbitrary unknown input state (not only a coherent state) can be restored in the other clone by applying appropriate local unitary displacement operation. We generalize this concept to a partial reversal of the cloning using only local operations and classical communication (LOCC) and we show that this procedure converts the symmetric cloner to an asymmetric cloner. Further, we discuss a distributed LOCC reversal in optimal 1{yields}M Gaussian cloning of coherent states which transforms it to optimal 1{yields}M{sup '} cloning for M{sup '}cloning as a possible eavesdropping attack on quantum communication link, the reversibility can be utilized to improve the security of the link even after the attack.

  9. Cloning of endangered mammalian species: any progress?

    PubMed

    Loi, Pasqualino; Galli, Cesare; Ptak, Grazyna

    2007-05-01

    Attempts through somatic cell nuclear transfer to expand wild populations that have shrunk to critical numbers is a logical extension of the successful cloning of mammals. However, although the first mammal was cloned 10 years ago, nuclear reprogramming remains phenomenological, with abnormal gene expression and epigenetic deregulation being associated with the cloning process. In addition, although cloning of wild animals using host oocytes from different species has been successful, little is known about the implication of partial or total mitochondrial DNA heteroplasmy in cloned embryos, fetuses and offspring. Finally, there is a need for suitable foster mothers for inter-intra specific cloned embryos. Considering these issues, the limited success achieved in cloning endangered animals is not surprising. However, optimism comes from the rapid gain in the understanding of the molecular clues underlying nuclear reprogramming. If it is possible to achieve a controlled reversal of the differentiated state of a cell then it is probable that other issues that impair the cloning of endangered animals, such as the inter-intra species oocyte or womb donor, will be overcome in the medium term. PMID:17379340

  10. CLONING AND EXPRESSION OF RABBIT INTERLEUKIN-15

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In order to understand the inflammatory mechanisms related to rabbit interleukin-15 (RIL-15), we cloned and expressed RIL-15 cDNA gene. A cDNA encoding RIL-15 was cloned from heart mRNA by reverse transcriptase polymerase chain reaction (RT-PCR) amplification using hIL-15 primers. The RIL-15 cDNA co...

  11. "Goodbye Dolly?" The ethics of human cloning.

    PubMed Central

    Harris, J

    1997-01-01

    The ethical implications of human clones have been much alluded to, but have seldom been examined with any rigour. This paper examines the possible uses and abuses of human cloning and draws out the principal ethical dimensions, both of what might be done and its meaning. The paper examines some of the major public and official responses to cloning by authorities such as President Clinton, the World Health Organisation, the European parliament, UNESCO, and others and reveals their inadequacies as foundations for a coherent public policy on human cloning. The paper ends by defending a conception of reproductive rights of "procreative autonomy" which shows human cloning to be not inconsistent with human rights and dignity. PMID:9451604

  12. Chorioallantoic placenta defects in cloned mice

    SciTech Connect

    Wakisaka-Saito, Noriko; Kohda, Takashi . E-mail: tkhoda.epgn@tmd.ac.jp; Inoue, Kimiko; Ogonuki, Narumi; Miki, Hiromi; Hikichi, Takafusa; Mizutani, Eiji; Wakayama, Teruhiko; Kaneko-Ishino, Tomoko; Ogura, Atsuo; Ishino, Fumitoshi

    2006-10-13

    Somatic cell nuclear transfer technology has been applied to produce live clones successfully in several mammalian species, but the success rates are very low. In mice, about half of the nuclear transfer embryos undergo implantation, but very few survive to term. We undertook detailed histological analyses of placentas from cloned mouse embryos generated from cumulus cells at 10.5 dpc of pregnancy, by which stage most clones have terminated their development. At 10.5 dpc, the extraembryonic tissues displayed several defined histological patterns, each reflecting their stage of developmental arrest. The most notable abnormality was the poor development of the spongiotrophoblast layer of diploid cells. This is in contrast to the placental hyperplasia frequently observed in somatic clones at 12.5 dpc or later stages. A variety of structural abnormalities were also observed in the embryos. Both placental and embryonic defects likely contribute to the low success rate of the mouse clones.

  13. Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for Differentiation of the Dimorphic Fungal Species Paracoccidioides brasiliensis and Paracoccidioides lutzii

    PubMed Central

    Del Negro, Gilda M. B.; Grenfell, Rafaella C.; Vidal, Monica S. M.; Thomaz, Danilo Y.; de Figueiredo, Dulce S. Y.; Bagagli, Eduardo; Juliano, Luiz; Benard, Gil

    2015-01-01

    Isolates of Paracoccidioides brasiliensis and Paracoccidioides lutzii, previously characterized by molecular techniques, were identified for the first time by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS). All isolates were correctly identified, with log score values of >2.0. Thus, MALDI-TOF MS is a new tool for differentiating species of the genus Paracoccidioides. PMID:25631803

  14. Brasiliquinones A, B and C, new benz[alpha]anthraquinone antibiotics from Nocardia brasiliensis. I. Producing strain, isolation and biological activities of the antibiotics.

    PubMed

    Nemoto, A; Tanaka, Y; Karasaki, Y; Komaki, H; Yazawa, K; Mikami, Y; Tojo, T; Kadowaki, K; Tsuda, M; Kobayashi, J

    1997-01-01

    New benz[alpha]anthraquinone antibiotics (brasiliquinones A, B and C) with an ethyl group at C-3 were isolated. The producer was identified as Nocardia brasiliensis. The antibiotics were active against Gram-positive bacteria including Mycobacterium sp., but not active against Gram-negative bacteria or fungi. They were also active against multiple drug-resistant P388/ADR tumor cells. PMID:9066761

  15. AQUA Cloning: A Versatile and Simple Enzyme-Free Cloning Approach

    PubMed Central

    Beyer, Hannes M.; Gonschorek, Patrick; Samodelov, Sophia L.; Meier, Matthias; Weber, Wilfried; Zurbriggen, Matias D.

    2015-01-01

    Assembly cloning is increasingly replacing conventional restriction enzyme and DNA-ligase-dependent cloning methods for reasons of efficiency and performance. Here, we describe AQUA (advanced quick assembly), a simple and versatile seamless assembly cloning approach. We demonstrate the applicability and versatility of AQUA Cloning in selected proof-of-principle applications including targeted insertion-, deletion- and site-directed point-mutagenesis, and combinatorial cloning. Furthermore, we show the one pot de novo assembly of multiple DNA fragments into a single circular plasmid encoding a complex light- and chemically-regulated Boolean A NIMPLY B logic operation. AQUA Cloning harnesses intrinsic in vivo processing of linear DNA fragments with short regions of homology of 16 to 32 bp mediated by Escherichia coli. It does not require any kits, enzymes or preparations of reagents and is the simplest assembly cloning protocol to date. PMID:26360249

  16. AQUA Cloning: A Versatile and Simple Enzyme-Free Cloning Approach.

    PubMed

    Beyer, Hannes M; Gonschorek, Patrick; Samodelov, Sophia L; Meier, Matthias; Weber, Wilfried; Zurbriggen, Matias D

    2015-01-01

    Assembly cloning is increasingly replacing conventional restriction enzyme and DNA-ligase-dependent cloning methods for reasons of efficiency and performance. Here, we describe AQUA (advanced quick assembly), a simple and versatile seamless assembly cloning approach. We demonstrate the applicability and versatility of AQUA Cloning in selected proof-of-principle applications including targeted insertion-, deletion- and site-directed point-mutagenesis, and combinatorial cloning. Furthermore, we show the one pot de novo assembly of multiple DNA fragments into a single circular plasmid encoding a complex light- and chemically-regulated Boolean A NIMPLY B logic operation. AQUA Cloning harnesses intrinsic in vivo processing of linear DNA fragments with short regions of homology of 16 to 32 bp mediated by Escherichia coli. It does not require any kits, enzymes or preparations of reagents and is the simplest assembly cloning protocol to date. PMID:26360249

  17. Antifungal activity of extracts from Atacama Desert fungi against Paracoccidioides brasiliensis and identification of Aspergillus felis as a promising source of natural bioactive compounds

    PubMed Central

    Mendes, Graziele; Gonçalves, Vívian N; Souza-Fagundes, Elaine M; Kohlhoff, Markus; Rosa, Carlos A; Zani, Carlos L; Cota, Betania B; Rosa, Luiz H; Johann, Susana

    2016-01-01

    Fungi of the genus Paracoccidioides are responsible for paracoccidioidomycosis. The occurrence of drug toxicity and relapse in this disease justify the development of new antifungal agents. Compounds extracted from fungal extract have showing antifungal activity. Extracts of 78 fungi isolated from rocks of the Atacama Desert were tested in a microdilution assay against Paracoccidioides brasiliensis Pb18. Approximately 18% (5) of the extracts showed minimum inhibitory concentration (MIC) values≤ 125.0 µg/mL. Among these, extract from the fungus UFMGCB 8030 demonstrated the best results, with an MIC of 15.6 µg/mL. This isolate was identified as Aspergillus felis (by macro and micromorphologies, and internal transcribed spacer, β-tubulin, and ribosomal polymerase II gene analyses) and was grown in five different culture media and extracted with various solvents to optimise its antifungal activity. Potato dextrose agar culture and dichloromethane extraction resulted in an MIC of 1.9 µg/mL against P. brasiliensis and did not show cytotoxicity at the concentrations tested in normal mammalian cell (Vero). This extract was subjected to bioassay-guided fractionation using analytical C18RP-high-performance liquid chromatography (HPLC) and an antifungal assay using P. brasiliensis. Analysis of the active fractions by HPLC-high resolution mass spectrometry allowed us to identify the antifungal agents present in the A. felis extracts cytochalasins. These results reveal the potential of A. felis as a producer of bioactive compounds with antifungal activity. PMID:27008375

  18. Antifungal activity of extracts from Atacama Desert fungi against Paracoccidioides brasiliensis and identification of Aspergillus felis as a promising source of natural bioactive compounds.

    PubMed

    Mendes, Graziele; Gonçalves, Vívian N; Souza-Fagundes, Elaine M; Kohlhoff, Markus; Rosa, Carlos A; Zani, Carlos L; Cota, Betania B; Rosa, Luiz H; Johann, Susana

    2016-03-01

    Fungi of the genus Paracoccidioides are responsible for paracoccidioidomycosis. The occurrence of drug toxicity and relapse in this disease justify the development of new antifungal agents. Compounds extracted from fungal extract have showing antifungal activity. Extracts of 78 fungi isolated from rocks of the Atacama Desert were tested in a microdilution assay against Paracoccidioides brasiliensis Pb18. Approximately 18% (5) of the extracts showed minimum inhibitory concentration (MIC) values ≤ 125.0 µg/mL. Among these, extract from the fungus UFMGCB 8030 demonstrated the best results, with an MIC of 15.6 µg/mL. This isolate was identified as Aspergillus felis (by macro and micromorphologies, and internal transcribed spacer, β-tubulin, and ribosomal polymerase II gene analyses) and was grown in five different culture media and extracted with various solvents to optimise its antifungal activity. Potato dextrose agar culture and dichloromethane extraction resulted in an MIC of 1.9 µg/mL against P. brasiliensis and did not show cytotoxicity at the concentrations tested in normal mammalian cell (Vero). This extract was subjected to bioassay-guided fractionation using analytical C18RP-high-performance liquid chromatography (HPLC) and an antifungal assay using P. brasiliensis. Analysis of the active fractions by HPLC-high resolution mass spectrometry allowed us to identify the antifungal agents present in the A. felis extracts cytochalasins. These results reveal the potential of A. felis as a producer of bioactive compounds with antifungal activity. PMID:27008375

  19. Therapeutic Administration of KM+ Lectin Protects Mice Against Paracoccidioides brasiliensis Infection via Interleukin-12 Production in a Toll-Like Receptor 2-Dependent Mechanism

    PubMed Central

    Coltri, Kely C.; Oliveira, Leandro L.; Pinzan, Camila F.; Vendruscolo, Patrícia E.; Martinez, Roberto; Goldman, Maria Helena; Panunto-Castelo, Ademilson; Roque-Barreira, Maria-Cristina

    2008-01-01

    KM+ is a mannose-binding lectin from Artocarpus integrifolia that induces interleukin (IL)-12 production by macrophages and protective T helper 1 immune response against Leishmania major infection. In this study, we performed experiments to evaluate the therapeutic activity of jackfruit KM+ (jfKM+) and its recombinant counterpart (rKM+) in experimental paracoccidioidomycosis. To this end, jfKM+ or rKM+ was administered to BALB/c mice 10 days after infection with Paracoccidiodes brasiliensis. Thirty days postinfection, lungs from the KM+-treated mice contained significantly fewer colony-forming units and little to no organized granulomas compared to the controls. In addition, lung homogenates from the KM+-treated mice presented higher levels of nitric oxide, IL-12, interferon-γ, and tumor necrosis factor-α, whereas higher levels of IL-4 and IL-10 were detected in the control group. With mice deficient in IL-12, Toll-like receptor (TLR) 2, TLR4, or TLR adaptor molecule MyD88, we demonstrated that KM+ led to protection against P. brasiliensis infection through IL-12 production, which was dependent on TLR2. These results demonstrated a beneficial effect of KM+ on the severity of P. brasiliensis infection and may expand its potential use as a novel immunotherapeutic molecule. PMID:18599609

  20. Protection against Paracoccidioides brasiliensis infection in mice treated with modulated dendritic cells relies on inhibition of interleukin-10 production by CD8+ T cells.

    PubMed

    da Costa, Thiago Alves; Di Gangi, Rosária; Martins, Paula; Longhini, Ana Leda Figueiredo; Zanucoli, Fábio; de Oliveira, Alexandre Leite Rodrigues; Stach-Machado, Dagmar Ruth; Burger, Eva; Verinaud, Liana; Thomé, Rodolfo

    2015-11-01

    Paracoccidioidomycosis is a systemic infection prevalent in Latin American countries. Disease develops after inhalation of Paracoccidioides brasiliensis conidia followed by an improper immune activation by the host leucocytes. Dendritic cells (DCs) are antigen-presenting cells with the unique ability to direct the adaptive immune response by the time of activation of naive T cells. This study was conducted to test whether extracts of P. brasiliensis would induce maturation of DCs. We found that DCs treated with extracts acquired an inflammatory phenotype and upon adoptive transfer conferred protection to infection. Interestingly, interleukin-10 production by CD8(+) T cells was ablated following DC transfer. Further analyses showed that lymphocytes from infected mice were high producers of interleukin-10, with CD8(+) T cells being the main source. Blockage of cross-presentation to CD8(+) T cells by modulated DCs abolished the protective effect of adoptive transfer. Collectively, our data show that adoptive transfer of P. brasiliensis-modulated DCs is an interesting approach for the control of infection in paracoccidioidomycosis. PMID:26302057

  1. Fluorescence in situ hybridization of 16S rRNA gene clones (Clone-FISH) for probe validation and screening of clone libraries.

    PubMed

    Schramm, Andreas; Fuchs, Bernhard M; Nielsen, Jeppe L; Tonolla, Mauro; Stahl, David A

    2002-11-01

    A method is presented for fluorescence in situ hybridization (FISH) of 16S rRNA gene clones targeting in vivo transcribed plasmid inserts (Clone-FISH). Several different cloning approaches and treatments to generate target-rRNA in the clones were compared. Highest signal intensities of Clone-FISH were obtained using plasmids with a T7 RNA polymerase promoter and host cells with an IPTG-inducible T7 RNA polymerase. Combined IPTG-induction and chloramphenicol treatment of those clones resulted in FISH signals up to 2.8-fold higher than signals of FISH with probe EUB338 to cells of Escherichia coli. Probe dissociation curves for three oligonucleotide probes were compared for reference cells containing native (FISH) or cloned (Clone-FISH) target sequences. Melting behaviour and calculated T(d) values were virtually identical for clones and cells, providing a format to use 16S rRNA gene clones instead of pure cultures for probe validation and optimization of hybridization conditions. The optimized Clone-FISH protocol was also used to screen an environmental clone library for insert sequences of interest. In this application format, 13 out of 82 clones examined were identified to contain sulphate-reducing bacterial rRNA genes. In summary, Clone-FISH is a simple and fast technique, compatible with a wide variety of cloning vectors and hosts, that should have general utility for probe validation and screening of clone libraries. PMID:12460279

  2. Who is the parent in cloning?

    PubMed

    Elster, N

    1999-01-01

    In July 1996, a sheep named Dolly was born in Scotland. What makes Dolly's birth noteworthy is that she is the result of the first successful cloning attempt using the nucleus of an adult cell. The technique that led to Dolly's birth involved transferring the nucleus of a mammary cell from an adult sheep to the enucleated egg cell of an unrelated sheep with gestation occurring in a third sheep. The possibility of applying this technique to human reproduction raised concerns worldwide with several countries moving for an immediate bans on human cloning. In the United States, President Clinton requested that the National Bioethics Advisory Commission ("NBAC"), a multidisciplinary group composed of scientists, lawyers, educators, theologians, and ethicists study the implications of cloning and issue recommendations. The Commission consulted other scientists, ethicists, theologians, lawyers, and citizens with interests in this advancing technology and concluded that, "at this time it is morally unacceptable for anyone in the public or private sector, whether in a research or clinical setting, to attempt to create a child using somatic cell nuclear transfer cloning." This Article was included in a larger work prepared at the request of, and submitted to the Commission by, law professor Lori B. Andrews. Cloning through nuclear transfer will change the way we create and define families. This Article explores how existing law relating to parentage, surrogacy, egg donation, and artificial insemination may apply in the cloning context to clarify the parent-child relationship established through cloning. PMID:12650149

  3. Economical quantum cloning in any dimension

    SciTech Connect

    Durt, Thomas; Fiurasek, Jaromir; Cerf, Nicolas J.

    2005-11-15

    The possibility of cloning a d-dimensional quantum system without an ancilla is explored, extending on the economical phase-covariant cloning machine for qubits found in Phys. Rev. A 60, 2764 (1999). We prove the impossibility of constructing an economical version of the optimal universal 1{yields}2 cloning machine in any dimension. We also show, using an ansatz on the generic form of cloning machines, that the d-dimensional 1{yields}2 phase-covariant cloner, which optimally clones all balanced superpositions with arbitrary phases, can be realized economically only in dimension d=2. The used ansatz is supported by numerical evidence up to d=7. An economical phase-covariant cloner can nevertheless be constructed for d>2, albeit with a slightly lower fidelity than that of the optimal cloner requiring an ancilla. Finally, using again an ansatz on cloning machines, we show that an economical version of the 1{yields}2 Fourier-covariant cloner, which optimally clones the computational basis and its Fourier transform, is also possible only in dimension d=2.

  4. Controlled secret sharing protocol using a quantum cloning circuit

    NASA Astrophysics Data System (ADS)

    Adhikari, Satyabrata; Roy, Sovik; Chakraborty, Shantanav; Jagadish, Vinayak; Haris, M. K.; Kumar, Atul

    2014-09-01

    We demonstrate the possibility of controlling the success probability of a secret sharing protocol using a quantum cloning circuit. The cloning circuit is used to clone the qubits containing the encoded information and en route to the intended recipients. The success probability of the protocol depends on the cloning parameters used to clone the qubits. We also establish a relation between the concurrence of initially prepared state, entanglement of the mixed state received by the receivers after cloning scheme and the cloning parameters of cloning machine.

  5. (New hosts and vectors for genome cloning)

    SciTech Connect

    Not Available

    1991-01-01

    The main goal of our project remains the development of new bacterial hosts and vectors for the stable propagation of human DNA clones in E. coli. During the past six months of our current budget period, we have (1) continued to develop new hosts that permit the stable maintenance of unstable features of human DNA, and (2) developed a series of vectors for (a) cloning large DNA inserts, (b) assessing the frequency of human sequences that are lethal to the growth of E. coli, and (c) assessing the stability of human sequences cloned in M13 for large-scale sequencing projects.

  6. Cloning and characterization of new bioluminescent proteins

    NASA Astrophysics Data System (ADS)

    Szent-Gyorgyi, Christopher; Ballou, Byron T.; Dagnal, Erich; Bryan, Bruce

    1999-07-01

    Over the past two years Prolume has undertaken a comprehensive program to clone luciferases and associated 'green fluorescent proteins' (GFPs) from marine animals that use coelenterazine as the luciferin. To data we have cloned several bioluminescent proteins, including two novel copepod luciferases and two anthozoan GFPs. These four proteins have sequences that differ greatly form previously cloned analogous proteins; the sequence diversity apparently is due to independent evolutionary origins and unusual evolutionary constraints. Thus coelenterazine-based bioluminescent systems may also manifest a variety of useful properties. We discuss form this taxonomic perspective the initial biochemical and spectral characterization of our cloned proteins. Emphasis is placed on the anthozoan luciferase-GFP systems, whose efficient resonance energy transfer has elicited much current interest.

  7. Optimal cloning of mixed Gaussian states

    SciTech Connect

    Guta, Madalin; Matsumoto, Keiji

    2006-09-15

    We construct the optimal one to two cloning transformation for the family of displaced thermal equilibrium states of a harmonic oscillator, with a fixed and known temperature. The transformation is Gaussian and it is optimal with respect to the figure of merit based on the joint output state and norm distance. The proof of the result is based on the equivalence between the optimal cloning problem and that of optimal amplification of Gaussian states which is then reduced to an optimization problem for diagonal states of a quantum oscillator. A key concept in finding the optimum is that of stochastic ordering which plays a similar role in the purely classical problem of Gaussian cloning. The result is then extended to the case of n to m cloning of mixed Gaussian states.

  8. Generation of phase-covariant quantum cloning

    SciTech Connect

    Karimipour, V.; Rezakhani, A.T.

    2002-11-01

    It is known that in phase-covariant quantum cloning, the equatorial states on the Bloch sphere can be cloned with a fidelity higher than the optimal bound established for universal quantum cloning. We generalize this concept to include other states on the Bloch sphere with a definite z component of spin. It is shown that once we know the z component, we can always clone a state with a fidelity higher than the universal value and that of equatorial states. We also make a detailed study of the entanglement properties of the output copies and show that the equatorial states are the only states that give rise to a separable density matrix for the outputs.

  9. Economical phase-covariant cloning with multiclones

    NASA Astrophysics Data System (ADS)

    Zhang, Wen-Hai; Ye, Liu

    2009-09-01

    This paper presents a very simple method to derive the explicit transformations of the optimal economical 1 to M phase-covariant cloning. The fidelity of clones reaches the theoretic bound [D'Ariano G M and Macchiavello C 2003 Phys. Rev. A 67 042306]. The derived transformations cover the previous contributions [Delgado Y, Lamata L et al., 2007 Phys. Rev. Lett. 98 150502] in which M must be odd.

  10. Intestinal mucosal mast cells from rats infected with Nippostrongylus brasiliensis contain protease-resistant chondroitin sulfate di-B proteoglycans

    SciTech Connect

    Stevens, R.L.; Lee, T.D.G.; Seldin, D.C.; Austen, K.F.; Befus, A.D.; Bienenstock, J.

    1986-07-01

    Rats infected with the helminth Nippostrongylus brasiliensis were injected i.p. with 2 mCi of (/sup 35/S) sulfate on days 13, 15, 17, and 19 after infection. The intestines were removed from animals on day 20 or 21 after infection, the intestinal cells were obtained by collagenase treatment and mechanical dispersion of the tissue, and the /sup 35/S-labeled mucosal mast cells (MMC) were enriched to 60 to 65% purity by Percoll centrifugation. The isolated proteoglycans were of approx. 150,000 m.w., were resistant to pronase degradation, and contained highly sulfated chondroitin sulfate side chains. The presence in normal mammalian cells of chondroitin sulfate proteoglycans that contain a high percentage of the unusual disulfated di-B disaccharide has not been previously reported. The rat intestinal MMC proteoglycans are the first chondroitin sulfate proteoglycans that have been isolated from an enriched populations of normal mast cells. They are homologous to the chondroitin sulfate-rich proteoglycans of the transformed rat basophilic leumekia-1 cell and the cultured interleukin 3-dependent mouse bone marrow-derived mast cell, in that these chondroitin sulfate proteoglycans are all highly sulfated, protease-resistant proteoglycans.

  11. Thyrotoxicosis followed by Hypothyroidism due to Suppurative Thyroiditis Caused by Nocardia brasiliensis in a Patient with Advanced Acquired Immunodeficiency Syndrome.

    PubMed

    Teckie, G2; Bhana, S A; Tsitsi, J M L; Shires, R

    2014-03-01

    Acute thyroiditis is an extremely rare complication of nocardiosis. We report a patient with hyperthyroidism due to suppurative thyroiditis caused by Nocardia brasiliensis. A 38-year-old Black male presented with features of thyrotoxicosis, sepsis and airway obstruction. He had no evidence of underlying thyroid disease, but was severely immunocompromised as a result of acquired immunodeficiency syndrome. He had previously been diagnosed with pulmonary nocardiosis and also had nocardial abscesses on his anterior chest wall. Investigations revealed thyrotoxicosis, with a FT4 of 43.2 pmol/l and a suppressed TSH <0.01 mIU/l. Serum anti-thyroperoxidase and anti-thyroglobulin antibodies were absent. Computed tomography scan showed a large abscess in the anterior neck involving the left lobe and isthmus, as well as inhomogeneous changes in the right lobe of the thyroid. The radioisotopic scan showed absent uptake of tracer in keeping with thyroiditis. Although the initial presentation was that of hyperthyroidism, destruction of the gland later resulted in sustained hypothyroidism, necessitating thyroid hormone supplementation. The hyperthyroidism can be explained by the release of presynthesized and stored thyroid hormone into the circulation as a result of inflammation and disruption of the thyroid follicles, and the subsequent hypothyroidism by the fact that much of the gland was destroyed by the abscess and the extensive inflammatory process. This is the first documented case of hyperthyroidism in a patient with acute suppurative thyroiditis caused by Nocardia. PMID:24847469

  12. 43-kilodalton glycoprotein from Paracoccidioides brasiliensis: immunochemical reactions with sera from patients with paracoccidioidomycosis, histoplasmosis, or Jorge Lobo's disease.

    PubMed Central

    Puccia, R; Travassos, L R

    1991-01-01

    Sera from patients with paracoccidioidomycosis (PCM), histoplasmosis (HP), or Jorge Lobo's disease (JL) were titrated against purified gp43 from Paracoccidioides brasiliensis by using both enzyme-linked immunosorbent assay (ELISA) and immunoprecipitation (IPP) reactions with 125I-labeled antigens. In IPP, PCM sera and other sera could be distinguished on the basis of serum titers, whereas in ELISA, 53% of the HP sera and 29% of the JL sera reacted similarly to the PCM sera. To investigate the possible role of the carbohydrate epitopes in these reactions, we compared the reactivities of sera from several patients with native and deglycosylated gp43. Competition experiments were carried out with monosaccharides as inhibitors. The results suggest that greater than 85% of the reactions of the PCM sera with gp43 involved peptide epitopes. Cross-reactions with HP and JL sera in ELISA were predominantly attributed to periodate-sensitive carbohydrate epitopes containing galactosyl residues. HP and JL sera which reacted strongly with gp43 in ELISA were only weakly reactive or did not react in IPP with labeled antigens in solution. Moreover, ELISA reactions could be significantly inhibited either by monosaccharides or by periodate treatment. Apparently, carbohydrate epitopes in gp43 are more accessible to the antibodies when the molecule is bound to a plastic substrate than when it is in solution. Structural changes in the gp43 antigen arising by N deglycosylation abolish reactivity with PCM sera and support the existence of conformational peptide epitopes. Images PMID:1722220

  13. Effects of sulfated and non-sulfated β-glucan extracted from Agaricus brasiliensis in breast adenocarcinoma cells - MCF-7.

    PubMed

    Baranoski, Adrivanio; Tempesta Oliveira, Marcelo; Semprebon, Simone Cristine; Niwa, Andressa Megumi; Ribeiro, Lúcia Regina; Mantovani, Mário Sérgio

    2015-11-01

    The β-glucans (β-G) are polysaccharides produced by various organisms, and sulfation of β-G renders them more soluble. With the objective to assess the effects of sulfated and non-sulfated β-G extracted from Agaricus brasiliensis in MCF-7 cells, assays were used to evaluate cytotoxicity, genotoxicity, cell proliferation and mRNA expression. The sulfated and non-sulfated β-G showed dose-dependent cytotoxicity at concentrations of 5 and 10 μg/mL, by the MTT assay. However, only cytotoxicity was observed after 24 h by the Red Neutral test for sulfated β-G, with no genotoxicity for either β-G in comet assay. Proliferation was decreased only at 72 h at a concentration of 100 μg/mL of sulfated β-G. Treatment with 5 μg/mL of sulfated β-G for 6 h reduced the expression of pro-apoptotic genes and stress signaling genes, cell cycle arrest, damage and cell migration. The 5 μg/mL of non-sulfated β-G for 6 h reduced the expression of the stress response gene and signaling damage. These results indicate that the cytotoxicity in the MTT is not cell death, and that, in general, sulfated β-G have greater cytotoxicity compared to non-sulfated β-G. PMID:25970150

  14. Anxiolytic Effects of Royal Sun Medicinal Mushroom, Agaricus brasiliensis (Higher Basidiomycetes) on Ischemia-Induced Anxiety in Rats.

    PubMed

    Zhang, Chunjing; Gao, Xiulan; Sun, Yan; Sun, Xiaojie; Wu, Yanmin; Liu, Ying; Yu, Haitao; Cui, Guangcheng

    2015-01-01

    We investigated the anxiolytic effects Agaricus brasiliensis extract (AbSE) on ischemia-induced anxiety using the plus-maze test and the social interaction test. The animals were treated orally with AbSE (4, 8, and 10 mg/kg/d, respectively) for 30 d, followed by middle cerebral artery occlusion-induced cerebral ischemia. Levels of noradrenaline, dopamine, and serotonin in the cerebral cortex of rats, as well as oxidative stress and plasma corticosterone levels were analyzed, respectively. The rota-rod test was carried out to exclude any false positive results in experimental procedures related to anxiety disorders, and the catalepsy test was carried out to investigate whether AbSE induces catalepsy. Our results demonstrate that oral administration of AbSE presented anxiolytic-like effects in the elevated plus-maze test and the social interaction test. Furthermore, AbSE did not induce extrapyramidal symptoms in the catalepsy test. The mechanism underlying the anxiolytic effect of AbSE might be increased brain monoamine levels and plasma corticosterone levels and decreased oxidative stress in cerebral ischemia/reperfusion rats. PMID:25746401

  15. Complete Genome Sequence Analysis of Nocardia brasiliensis HUJEG-1 Reveals a Saprobic Lifestyle and the Genes Needed for Human Pathogenesis

    PubMed Central

    Vera-Cabrera, Lucio; Ortiz-Lopez, Rocio; Elizondo-Gonzalez, Ramiro; Ocampo-Candiani, Jorge

    2013-01-01

    Nocardia brasiliensis is an important etiologic agent of mycetoma. These bacteria live as a saprobe in soil or organic material and enter the tissue via minor trauma. Mycetoma is characterized by tumefaction and the production of fistula and abscesses, with no spontaneous cure. By using mass sequencing, we determined the complete genomic nucleotide sequence of the bacteria. According to our data, the genome is a circular chromosome 9,436,348-bp long with 68% G+C content that encodes 8,414 proteins. We observed orthologs for virulence factors, a higher number of genes involved in lipid biosynthesis and catabolism, and gene clusters for the synthesis of bioactive compounds, such as antibiotics, terpenes, and polyketides. An in silico analysis of the sequence supports the conclusion that the bacteria acquired diverse genes by horizontal transfer from other soil bacteria, even from eukaryotic organisms. The genome composition reflects the evolution of bacteria via the acquisition of a large amount of DNA, which allows it to survive in new ecological niches, including humans. PMID:23755230

  16. Quillaja brasiliensis saponins induce robust humoral and cellular responses in a bovine viral diarrhea virus vaccine in mice.

    PubMed

    Cibulski, Samuel Paulo; Silveira, Fernando; Mourglia-Ettlin, Gustavo; Teixeira, Thais Fumaco; dos Santos, Helton Fernandes; Yendo, Anna Carolina; de Costa, Fernanda; Fett-Neto, Arthur Germano; Gosmann, Grace; Roehe, Paulo Michel

    2016-04-01

    A saponin fraction extracted from Quillaja brasiliensis leaves (QB-90) and a semi-purified aqueous extract (AE) were evaluated as adjuvants in a bovine viral diarrhea virus (BVDV) vaccine in mice. Animals were immunized on days 0 and 14 with antigen plus either QB-90 or AE or an oil-adjuvanted vaccine. Two-weeks after boosting, antibodies were measured by ELISA; cellular immunity was evaluated by DTH, lymphoproliferation, cytokine release and single cell IFN-γ production. Serum anti-BVDV IgG, IgG1 and IgG2b were significantly increased in QB-90- and AE-adjuvanted vaccines. A robust DTH response, increased splenocyte proliferation, Th1-type cytokines and enhanced production of IFN-γ by CD4(+) and CD8(+) T lymphocytes were detected in mice that received QB-90-adjuvanted vaccine. The AE-adjuvanted preparation stimulated humoral responses but not cellular immune responses. These findings reveal that QB-90 is capable of stimulating both cellular and humoral immune responses when used as adjuvant. PMID:27012913

  17. Novel ISCOMs from Quillaja brasiliensis saponins induce mucosal and systemic antibody production, T-cell responses and improved antigen uptake.

    PubMed

    Cibulski, Samuel Paulo; Mourglia-Ettlin, Gustavo; Teixeira, Thais Fumaco; Quirici, Lenora; Roehe, Paulo Michel; Ferreira, Fernando; Silveira, Fernando

    2016-02-24

    In the last decades, significant efforts have been dedicated to the search for novel vaccine adjuvants. In this regard, saponins and its formulations as "immunostimulating complexes" (ISCOMs) have shown to be capable of stimulating potent humoral and cellular immune responses, enhanced cytokine production and activation of cytotoxic T cells. The immunological activity of ISCOMs formulated with a saponin fraction extracted from Quillaja brasiliensis (QB-90 fraction) as an alternative to classical ISCOMs based on Quil A(®) (IQA) is presented here. The ISCOMs prepared with QB-90, named IQB-90, typically consist of 40-50 nm, spherical, cage-like particles, built up by QB-90, cholesterol, phospholipids and antigen (ovalbumin, OVA). These nanoparticles were efficiently uptaken in vitro by murine bone marrow-derived dendritic cells. Subcutaneously inoculated IQB-90 induced strong serum antibody responses encompassing specific IgG1 and IgG2a, robust DTH reactions, significant T cell proliferation and increases in Th1 (IFN-γ and IL-2) cytokine responses. Intranasally delivered IQB-90 elicited serum IgG and IgG1, and mucosal IgA responses at distal systemic sites (nasal passages, large intestine and vaginal lumen). These results indicate that IQB-90 is a promising alternative to classic ISCOMs as vaccine adjuvants, capable of enhancing humoral and cellular immunity to levels comparable to those induced by ISCOMs manufactured with Quillaja saponaria saponins. PMID:26826546

  18. Mucin-Related Molecular Responses of Bronchial Epithelial Cells in Rats Infected with the Nematode Nippostrongylus brasiliensis

    PubMed Central

    Soga, Koichi; Yamada, Minoru; Naito, Yuji; Yoshikawa, Toshikazu; Arizono, Naoki

    2013-01-01

    Although mucins are essential for the protection of internal epithelial surfaces, molecular responses involving mucin production and secretion in response to various infectious agents in the airway have not been fully elucidated. The present study analysed airway goblet cell mucins in rats infected with the nematode Nippostrongylus brasiliensis, which migrates to the lungs shortly after infection. Goblet cell hyperplasia occurred in the bronchial epithelium 3–10 days after infection. The high iron diamine-alcian blue staining combined with neuraminidase treatment showed that sialomucin is the major mucin in hyperplastic goblet cells. Immunohistochemical studies demonstrated that goblet cell mucins were immunoreactive with both the major airway mucin core peptide, Muc5AC, and the major intestinal mucin core peptide Muc2. Reverse transcription real-time PCR studies demonstrated upregulation of gene transcription levels of Muc5AC, Muc2, the sialyltransferase St3gal4, and the resistin-like molecule beta (Retnlb) in the lungs. These results showed that nematode infection induces airway epithelial responses characterised by the production of sialomucin with Muc5AC and Muc2 core peptides. These mucins, as well as Retnlb, might have important roles in the protection of mucosa from migrating nematodes in the airway. PMID:27335862

  19. The combined use of Paracoccidioides brasiliensis Pb40 and Pb27 recombinant proteins enhances chemotherapy effects in experimental paracoccidioidomycosis.

    PubMed

    Fernandes, Viviane C; Martins, Estefânia M N; Boeloni, Jankerle N; Coitinho, Juliana B; Serakides, Rogéria; Goes, Alfredo M

    2011-11-01

    Paracoccidioides brasiliensis is the etiological agent of paracoccidioidomycosis (PCM), a chronic granulomatous mycosis prevalent in Latin America, and cell-mediated immunity represents the main mode of protection against this fungal infection. The conventional treatment for this mycosis involves long periods of therapy resulting in sequels and a high frequency of relapse. The search for new alternative methods of treatment is thus necessary. With this aim, the objective of this work was to evaluate the potential of rPb27 and rPb40 immunization to reduce treatment length and the frequency of relapse when used as an adjuvant to fluconazole chemotherapy in experimental PCM. Combined treatment with the drug and the two proteins reduced CFUs in the lung, liver and spleen to undetectable levels and largely preserved the tissue structure of these organs. At the same time, IFN-γ and TNF-α levels were higher in mice treated as described above than in infected-only mice, while very low production of IL-10 and TGF-β was observed in this treated group. Thus, the combined treatment, using immunization with the two recombinant proteins in addition to fluconazole chemotherapy, showed an additive protective effect after intratracheal challenge. These results provide new prospects for immunotherapy as a treatment for PCM. PMID:21726659

  20. Evaluation of seasonal chemical composition, antibacterial, antioxidant and anticholinesterase activity of essential oil from Eugenia brasiliensis Lam.

    PubMed

    Siebert, Diogo Alexandre; Tenfen, Adrielli; Yamanaka, Celina Noriko; de Cordova, Caio Maurício Mendes; Scharf, Dilamara Riva; Simionatto, Edésio Luiz; Alberton, Michele Debiasi

    2015-02-01

    This study describes the seasonal composition and the antibacterial, antioxidant and anticholinesterase activity of the essential oil from Eugenia brasiliensis leaves. Analysis by using GC allowed the identification of 40 compounds. It was observed that the monoterpenes varied more (42%) than the sesquiterpenes (14%), and that the monoterpene hydrocarbons suffered the greatest variation throughout the year (64%). Major compounds were spathulenol in the spring (16.02 ± 0.44%) and summer (18.17 ± 0.41%), τ-cadinol in the autumn (12.83 ± 0.03%) and α-pinene (15.94 ± 0.58%) in the winter. Essential oils were tested for their antibacterial activity, and the best result was obtained from the autumn oil, with MIC = 500 μg mL(- 1) against Staphylococcus saprophyticus and Pseudomonas aeruginosa. Antioxidant activity was evaluated using DPPH, lipid peroxidation and iron-reducing power assays, as well as the anticholinesterase activity. Both tests showed a weak performance of the essential oils. PMID:25219800

  1. Cloning: Past, Present, and the Exciting Future. Breakthroughs in Bioscience.

    ERIC Educational Resources Information Center

    Di Berardino, Marie A.

    This document explores the history of cloning by focusing on Dolly the Sheep, one of the first large animal clonings. The disadvantages and advantages of transgenic clones are discussed as well as the future implications of cloning from the perspective of human health. (Contains 10 resources.) (YDS)

  2. Cloning the entanglement of a pair of quantum bits

    SciTech Connect

    Lamoureux, Louis-Philippe; Navez, Patrick; Cerf, Nicolas J.; Fiurasek, Jaromir

    2004-04-01

    It is shown that any quantum operation that perfectly clones the entanglement of all maximally entangled qubit pairs cannot preserve separability. This 'entanglement no-cloning' principle naturally suggests that some approximate cloning of entanglement is nevertheless allowed by quantum mechanics. We investigate a separability-preserving optimal cloning machine that duplicates all maximally entangled states of two qubits, resulting in 0.285 bits of entanglement per clone, while a local cloning machine only yields 0.060 bits of entanglement per clone.

  3. HbIDI, SlIDI and EcIDI: A comparative study of isopentenyl diphosphate isomerase activity and structure.

    PubMed

    Berthelot, Karine; Estevez, Yannick; Quiliano, Miguel; Baldera-Aguayo, Pedro A; Zimic, Mirko; Pribat, Anne; Bakleh, Marc-Elias; Teyssier, Emeline; Gallusci, Philippe; Gardrat, Christian; Lecomte, Sophie; Peruch, Frédéric

    2016-08-01

    In this study, we cloned, expressed and purified the isopentenyl diphosphate isomerases (IDIs) from two plants, Hevea brasiliensis and Solanum lycopersicum, and compared them to the already well characterized Escherichia coli IDI. Phylogenetic analysis showed high homology between the three enzymes. Their catalytic activity was investigated in vitro with recombinant purified enzymes and in vivo by complementation colorimetric tests. The three enzymes displayed consistent activities both in vitro and in vivo. In term of structure, studied by ATR-FTIR and molecular modeling, it is clear that both plant enzymes are more related to their human homologue than to E. coli IDI. But it is assumed that EcIDI represent the minimalistic part of the catalytic core, as both plant enzymes present a supplementary sequence forming an extra α-helice surrounding the catalytic site that could facilitate the biocatalysis. New potential biotechnological applications may be envisaged. PMID:27163845

  4. Benefits and problems with cloning animals.

    PubMed Central

    Smith, L C; Bordignon, V; Babkine, M; Fecteau, G; Keefer, C

    2000-01-01

    Animal cloning is becoming a useful technique for producing transgenic farm animals and is likely to be used to produce clones from valuable adults. Other applications will also undoubtedly be discovered in the near future, such as for preserving endangered breeds and species. Although cloning promises great advantages for commerce and research alike, its outcome is not always certain due to high pregnancy losses and high morbidity and mortality during the neonatal period. Research into the mechanisms involved in the reprogramming of the nucleus is being conducted throughout the world in an attempt to better understand the molecular and cellular mechanisms involved in correcting these problems. Although the cause of these anomalies remains mostly unknown, similar phenotypes have been observed in calves derived through in vitro fertilization, suggesting that culture conditions are involved in these phenomena. In the meantime, veterinarians and theriogenologists have an important role to play in improving the efficiency of cloning by finding treatments to assure normal gestation to term and to develop preventative and curative care for cloned neonates. Images Figure 1. PMID:11143925

  5. Modeling Disease Vector Occurrence When Detection Is Imperfect II: Drivers of Site-Occupancy by Synanthropic Triatoma brasiliensis in the Brazilian Northeast

    PubMed Central

    Valença-Barbosa, Carolina; Lima, Marli M.; Sarquis, Otília; Bezerra, Claudia M.; Abad-Franch, Fernando

    2014-01-01

    Background Understanding the drivers of habitat selection by insect disease vectors is instrumental to the design and operation of rational control-surveillance systems. One pervasive yet often overlooked drawback of vector studies is that detection failures result in some sites being misclassified as uninfested; naïve infestation indices are therefore biased, and this can confound our view of vector habitat preferences. Here, we present an initial attempt at applying methods that explicitly account for imperfect detection to investigate the ecology of Chagas disease vectors in man-made environments. Methodology We combined triplicate-sampling of individual ecotopes (n = 203) and site-occupancy models (SOMs) to test a suite of pre-specified hypotheses about habitat selection by Triatoma brasiliensis. SOM results were compared with those of standard generalized linear models (GLMs) that assume perfect detection even with single bug-searches. Principal Findings Triatoma brasiliensis was strongly associated with key hosts (native rodents, goats/sheep and, to a lesser extent, fowl) in peridomestic environments; ecotope structure had, in comparison, small to negligible effects, although wooden ecotopes were slightly preferred. We found evidence of dwelling-level aggregation of infestation foci; when there was one such focus, same-dwelling ecotopes, whether houses or peridomestic structures, were more likely to become infested too. GLMs yielded negatively-biased covariate effect estimates and standard errors; both were, on average, about four times smaller than those derived from SOMs. Conclusions/Significance Our results confirm substantial population-level ecological heterogeneity in T. brasiliensis. They also suggest that, at least in some sites, control of this species may benefit from peridomestic rodent control and changes in goat/sheep husbandry practices. Finally, our comparative analyses highlight the importance of accounting for the various sources of

  6. Hitting an Unintended Target: Phylogeography of Bombus brasiliensis Lepeletier, 1836 and the First New Brazilian Bumblebee Species in a Century (Hymenoptera: Apidae)

    PubMed Central

    Santos Júnior, José Eustáquio; Santos, Fabrício R.; Silveira, Fernando A.

    2015-01-01

    This work tested whether or not populations of Bombus brasiliensis isolated on mountain tops of southeastern Brazil belonged to the same species as populations widespread in lowland areas in the Atlantic coast and westward along the Paraná-river valley. Phylogeographic and population genetic analyses showed that those populations were all conspecific. However, they revealed a previously unrecognized, apparently rare, and potentially endangered species in one of the most threatened biodiversity hotspots of the World, the Brazilian Atlantic Forest. This species is described here as Bombus bahiensis sp. n., and included in a revised key for the identification of the bumblebee species known to occur in Brazil. Phylogenetic analyses based on two mtDNA markers suggest this new species to be sister to B. brasiliensis, from which its workers and queens can be easily distinguished by the lack of a yellow hair-band on the first metasomal tergum. The results presented here are consistent with the hypothesis that B. bahiensis sp. n. may have originated from an ancestral population isolated in an evergreen-forest refuge (the so-called Bahia refuge) during cold, dry periods of the Pleistocene. This refuge is also known as an important area of endemism for several animal taxa, including other bees. Secondary contact between B. bahiensis and B. brasiliensis may be presently prevented by a strip of semi-deciduous forest in a climate zone characterized by relatively long dry seasons. Considering the relatively limited range of this new species and the current anthropic pressure on its environment, attention should be given to its conservation status. PMID:25992624

  7. Marking Triatoma brasiliensis, Triatoma pseudomaculata and Rhodnius nasutus Nymphs with Trace Elements: Element Persistence and Effects of Marking on Insect Mortality

    PubMed Central

    Valença-Barbosa, Carolina; Sarquis, Otília; Freire, Aline Soares; David, Mariana R.; Santelli, Ricardo E.; Monteiro, Fernando A.; Lima, Marli M.; Maciel-de-Freitas, Rafael

    2016-01-01

    Background Field ecologists often rely on mark-release-recapture (MRR) experiments to estimate population dynamics parameters for a given species. In the case of a medically important taxon, i.e., a disease vector, inferences on species survival and dispersal rates are particularly important as they have the potential to provide insights into disease transmission dynamics in endemic areas. Medical entomologists have traditionally used fluorescent dusts to externally mark the cuticle of insects. However, dust marking is usually restricted to the adult life stage because immature insects lose the mark when they molt. Methodology/Principal Findings We evaluated the efficacy of 13 trace elements in marking nymphs of three native Brazilian Chagas disease vectors: Triatoma brasiliensis, Triatoma pseudomaculata, and Rhodnius nasutus. Cr and Cu were detected in over 97% of T. brasiliensis (34/35 31/31 for Cr and Cu), while Cu and Mn were detected in more than 95% of T. pseudomaculata (29/29 for Cu and 28/29 for Mn) tested 120 days after marking. Only Mn marked over 90% of R. nasutus nymphs (38/41). Overall, trace elements had no negative effects on T. pseudomaculata longevity, but As-marked T. brasiliensis nymphs (p<0.01), and Cd-marked R. nasutus nymphs (p<0.01) had significantly shorter lifespan. Conclusions/Significance Previous evidence shows that there is little or no genetic differentiation between populations at the microgeographic level, which often precludes indirect estimations of dispersal capability based on genetic markers. In such situations, MRR studies are more suitable as they measure insect movement directly from one site to another, instead of effective migration (i.e. gene flow). The determination of a reliable and persistent marking method is the first step towards the development of meaningful ecological estimates through the application of MRR methodology. Here, we have identified trace elements that can be used for mark and recapture studies of

  8. Nippostrongylus brasiliensis infection in the rat: effect of iron and protein deficiency on the anthelmintic efficacy of mebendazole, pyrantel, piperazine, and levamisole.

    PubMed Central

    Duncombe, V M; Bolin, T D; Davis, A E; Fagan, M R; Kelly, J D

    1979-01-01

    The benzimidazole anthelmintics mebendazole and fenbendazole have been shown to be much less effective against Nippostrongylus brasiliensis infections in the rat on a combined iron and protein deficient diet. In the present experiments it was shown that the anthelmintic efficacy of mebendazole was significantly impaired in the rat on either an iron deficient or a protein deficient diet. Furthermore, iron and protein deficiency reduced the efficacy of the anthelmintics pyrantel and piperazine but not levamisole. The finding that nutritional deficiencies reduce anthelmintic efficacy may well be relevant to worm eradication programmes in iron deficient and protein calorie malnourished populations. PMID:447110

  9. Methods in molecular cardiology: in silico cloning

    PubMed Central

    Passier, R.; Doevendans, P.A.

    2004-01-01

    Advancements in sequencing technology have made it possible to obtain more information about the DNA sequence, structure and the transcript products of the genome from different species. This information is collected in DNA databases. These databases contain many genes of which the functions have not yet been discovered. By using online biotechnology tools novel genes and their transcripts can be identified. The identification of novel genes using DNA database analysis is referred to as in silico cloning. In silico cloning may not only provide new information on genes and their biological function, it may also lead to identification of molecular targets for drug discovery activities. In this review we describe the process of in silico cloning and its application in biomedical research. ImagesFigure 1Figure 3 PMID:25696371

  10. Bounds for state-dependent quantum cloning

    SciTech Connect

    Han Yongjian; Zhang Yongsheng; Guo Guangcan

    2002-11-01

    Due to the no-cloning theorem, the unknown quantum state can only be cloned approximately or exactly with some probability. There are two types of cloners: universal and state-dependent cloner. The optimal universal cloner has been found and can be viewed as a special state-dependent quantum cloner that has no information about the states. In this paper, we investigate the state-dependent cloning when the state set contains more than two states. We get some bounds of the global fidelity for these processes. This method is not dependent on the number of the states contained in the state set. It is also independent of the numbers of copying.

  11. Cloning quantum entanglement in arbitrary dimensions

    SciTech Connect

    Karpov, E.; Navez, P.; Cerf, N.J.

    2005-10-15

    We have found a quantum cloning machine that optimally duplicates the entanglement of a pair of d-dimensional quantum systems prepared in an arbitrary isotropic state. It maximizes the entanglement of formation contained in the two copies of any maximally entangled input state, while preserving the separability of unentangled input states. Moreover, it cannot increase the entanglement of formation of isotropic states. For large d, the entanglement of formation of each clone tends to one-half the entanglement of the input state, which corresponds to a classical behavior. Finally, we investigate a local entanglement cloner, which yields entangled clones with one-fourth the input entanglement in the large-d limit.

  12. Bac clones generated from sheared dna

    SciTech Connect

    Osoegawa, Kazutoyo; Vessere, Gery M.; Shu, Chung Li; Hoskins,Roger A.; Abad, Jose P.; de Pablos, Beatriz; Villasante, Alfredo; deJong, Pieter J.

    2006-08-09

    BAC libraries generated from restriction-digested genomic DNA display representational bias and lack some sequences. To facilitate completion of genome projects, procedures have been developed to create BACs from DNA physically sheared to create fragments extending up to 200kb. The DNA fragments were repaired to create blunt ends and ligated to a new BAC vector. This approach has been tested by generating BAC libraries from Drosophila DNA, with insert lengths of 50 kb to 150 kb. The libraries lack chimeric clone problems as determined by mapping paired BAC-end sequences of one library to the D. melanogaster genome sequence. The utility of ''sheared'' libraries was demonstrated by closure of a previous clone gap and by isolation of clones from telomeric regions, which were notably absent from previous Drosophila BAC libraries.

  13. Photonic Programmable Tele-Cloning Network

    PubMed Central

    Li, Wei; Chen, Ming-Cheng

    2016-01-01

    The concept of quantum teleportation allows an unknown quantum states to be broadcasted and processed in a distributed quantum network. The quantum information injected into the network can be diluted to distant multi-copies by quantum cloning and processed by arbitrary quantum logic gates which were programed in advance in the network quantum state. A quantum network combines simultaneously these fundamental quantum functions could lead to new intriguing applications. Here we propose a photonic programmable telecloning network based on a four-photon interferometer. The photonic network serves as quantum gate, quantum cloning and quantum teleportation and features experimental advantage of high brightness by photon recycling. PMID:27353838

  14. [MRSA clones identified in outpatient dermatology clinics].

    PubMed

    Hosoya, Shino; Ito, Teruyo; Misawa, Shigeki; Yoshiike, Takashi; Oguri, Toyoko; Hiramatsu, Keiichi

    2014-11-01

    To know the characteristics of methicillin-resistant Staphylococcus aureus (MRSA) strains disseminating through the Japanese community, we have determined types of Staphylococcal cassette chromosome mec (SCCmec) elements, Multi-Locus Sequence Typing (MLST), and carriages of four exotoxin genes (toxic-shock syndrome toxin, Panton-Valentine Leukocidine, and exfoliative toxins a and b) using 54 MRSA strains isolated from outpatients attending dermatology clinics at the four university hospitals of Juntendo University. Ten clonal complexes and 12 SCCmec types have been identified. As a result, more than 15 MRSA clones that were defined by the combination of genotype and SCCmec type, were identified. Among them, Clonal Complex (CC) 5-type IIa SCCmec strains were the most major (16 strains). In contrast to the fact that CC5- type IIa SCCmec strains known as a hospital-associated MRSA clone in Japan carried toxic-shock syndrome toxin gene (tst), only 2 of 16 strains have been shown to carry tst. Thirty-eight (70.4%) of isolates belonged to the clones distinct from the CC5-type IIa SCCmec strains. Among them, CC8 strains were major (12 strains), which contained 9 tst-positive CC8-type IVl SCCmec clones and a CC8-type IVa SCCmec strain carrying the Panton Valentine Leukocidin gene (lukS, F-PV). Clones related to impetigo were also identified: 7 exfoliative toxin b (etb) -positive clones, CC89-type IIa SCCmec and CC89-type V SCCmec strains; and 2 exfoliative toxin a (eta) -positive CC121-type V SCCmec strains. Other clones were as follows: CC1-type IVa SCCmec, CC8-type I SCCmec, CC81-type IVg SCCmec, CC97-type IVc SCCmec, CC91-type IVa SCCmec, CC59-type IVg SCCmec, CC45-type IIn SCCmec, CC89-SCCmec nontypeable, and CC8-type IVm, novel subtype of type IV SCCmec were identified in this study. Our data showed that many novel MRSA clones have emerged in the community. PMID:25764806

  15. Photonic Programmable Tele-Cloning Network.

    PubMed

    Li, Wei; Chen, Ming-Cheng

    2016-01-01

    The concept of quantum teleportation allows an unknown quantum states to be broadcasted and processed in a distributed quantum network. The quantum information injected into the network can be diluted to distant multi-copies by quantum cloning and processed by arbitrary quantum logic gates which were programed in advance in the network quantum state. A quantum network combines simultaneously these fundamental quantum functions could lead to new intriguing applications. Here we propose a photonic programmable telecloning network based on a four-photon interferometer. The photonic network serves as quantum gate, quantum cloning and quantum teleportation and features experimental advantage of high brightness by photon recycling. PMID:27353838

  16. Cloning of Gaussian states by linear optics

    SciTech Connect

    Olivares, Stefano; Paris, Matteo G. A.; Andersen, Ulrik L.

    2006-06-15

    We analyze in details a scheme for cloning of Gaussian states based on linear optical components and homodyne detection recently demonstrated by Andersen et al. [Phys. Rev. Lett. 94, 240503 (2005)]. The input-output fidelity is evaluated for a generic (pure or mixed) Gaussian state taking into account the effect of nonunit quantum efficiency and unbalanced mode mixing. In addition, since in most quantum information protocols the covariance matrix of the set of input states is not perfectly known, we evaluate the average cloning fidelity for classes of Gaussian states with the degree of squeezing and the number of thermal photons being only partially known.

  17. Photonic Programmable Tele-Cloning Network

    NASA Astrophysics Data System (ADS)

    Li, Wei; Chen, Ming-Cheng

    2016-06-01

    The concept of quantum teleportation allows an unknown quantum states to be broadcasted and processed in a distributed quantum network. The quantum information injected into the network can be diluted to distant multi-copies by quantum cloning and processed by arbitrary quantum logic gates which were programed in advance in the network quantum state. A quantum network combines simultaneously these fundamental quantum functions could lead to new intriguing applications. Here we propose a photonic programmable telecloning network based on a four-photon interferometer. The photonic network serves as quantum gate, quantum cloning and quantum teleportation and features experimental advantage of high brightness by photon recycling.

  18. Demography of the giant otter (Pteronura brasiliensis) in Manu National Park, south-eastern Peru: implications for conservation.

    PubMed

    Groenendijk, Jessica; Hajek, Frank; Johnson, Paul J; Macdonald, David W; Calvimontes, Jorge; Staib, Elke; Schenck, Christof

    2014-01-01

    The giant otter (Pteronura brasiliensis) is an endangered semi-aquatic carnivore of South America. We present findings on the demography of a population inhabiting the floodplain of Manu National Park, south-eastern Peru, arising from 14 annual dry season censuses over a 16 year period. The breeding system of territorial groups, including only a single breeding female with non-reproductive adult 'helpers', resulted in a low intrinsic rate of increase (0.03) and a slow recovery from decades of hunting for the pelt trade. This is explained by a combination of factors: (1) physiological traits such as late age at first reproduction and long generation time, (2) a high degree of reproductive skew, (3) small litters produced only once a year, and (4) a 50% mortality between den emergence and age of dispersal, as well as high mortality amongst dispersers (especially males). Female and male giant otters show similar traits with respect to average reproductive life-spans (female 5.4 yrs., male 5.2 yrs.) and average cub productivity (female 6.9, male 6.7 cubs per lifetime); the longest reproductive life spans were 11 and 13 years respectively. Individual reproductive success varied substantially and depended mainly on the duration of dominance tenure in the territory. When breeding females died, the reproductive position in the group was usually occupied by sisters or daughters (n = 11), with immigrant male partners. Male philopatry was not observed. The vulnerability of the Manu giant otter population to anthropogenic disturbance emphasises the importance of effective protection of core lake habitats in particular. Riverine forests are the most endangered ecosystem in the Department of Madre de Dios due to the concentration of gold mining, logging and agricultural activities in floodplains, highlighting the need for a giant otter habitat conservation corridor along the Madre de Dios River. PMID:25162684

  19. Oxidative Status Profile in Different Tissues of Parastacus brasiliensis promatensis (Crustacea, Decapoda, Parastacidae) over a Seasonal Cycle.

    PubMed

    Pinheiro, Ludimila Carneiro; Oliveira, Guendalina Turcato

    2016-06-01

    The objective of this study was to assess annual redox balance by quantifying activity of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST), as well as measuring lipid peroxidation levels by determination of thiobarbituric acid-reactive substances, in different tissues of male and female crayfish collected from an area little affected by anthropogenic impacts. A total of 37 male and 32 female of Parastacus brasiliensis promatensis were collected over an annual cycle, and gill, muscle, hepatopancreas, and gonad samples were harvested and subjected to the aforementioned analyses via spectrophotometry. Comparison of the annual oxidative status response profile in gill and in hepatopancreas between males and females revealed differences only in SOD activity. In abdominal muscle, this comparison revealed a distinct profile of SOD and GST activity, as well as of lipid peroxidation. SOD activity in gonads of females increased in the summer when compared to autumn; conversely, CAT and GST levels did not differ over the year. During reproduction, a possible increase of the energy requirements led to an increase in lipid peroxidation in all tested tissues, in males and females alike. It was least evident in female gonad tissue, which suggests this tissue type is relatively protected; within this context, one may establish a tissue-specific grading of Lipid Peroxidation intensity in females of the species during the spring: gonads < muscle < gills < hepatopancreas. Our results show that the responses of the defenses analyzed antioxidants, as well as lipid peroxidation levels, were sex and tissue dependent and a clear pattern of seasonal variation. PMID:27256498

  20. Demography of the Giant Otter (Pteronura brasiliensis) in Manu National Park, South-Eastern Peru: Implications for Conservation

    PubMed Central

    Groenendijk, Jessica; Hajek, Frank; Johnson, Paul J.; Macdonald, David W.; Calvimontes, Jorge; Staib, Elke; Schenck, Christof

    2014-01-01

    The giant otter (Pteronura brasiliensis) is an endangered semi-aquatic carnivore of South America. We present findings on the demography of a population inhabiting the floodplain of Manu National Park, south-eastern Peru, arising from 14 annual dry season censuses over a 16 year period. The breeding system of territorial groups, including only a single breeding female with non-reproductive adult ‘helpers’, resulted in a low intrinsic rate of increase (0.03) and a slow recovery from decades of hunting for the pelt trade. This is explained by a combination of factors: (1) physiological traits such as late age at first reproduction and long generation time, (2) a high degree of reproductive skew, (3) small litters produced only once a year, and (4) a 50% mortality between den emergence and age of dispersal, as well as high mortality amongst dispersers (especially males). Female and male giant otters show similar traits with respect to average reproductive life-spans (female 5.4 yrs., male 5.2 yrs.) and average cub productivity (female 6.9, male 6.7 cubs per lifetime); the longest reproductive life spans were 11 and 13 years respectively. Individual reproductive success varied substantially and depended mainly on the duration of dominance tenure in the territory. When breeding females died, the reproductive position in the group was usually occupied by sisters or daughters (n = 11), with immigrant male partners. Male philopatry was not observed. The vulnerability of the Manu giant otter population to anthropogenic disturbance emphasises the importance of effective protection of core lake habitats in particular. Riverine forests are the most endangered ecosystem in the Department of Madre de Dios due to the concentration of gold mining, logging and agricultural activities in floodplains, highlighting the need for a giant otter habitat conservation corridor along the Madre de Dios River. PMID:25162684

  1. Alterations in the pathogenicity of one Paracoccidioides brasiliensis isolate do not correlative with its in vitro growth.

    PubMed

    Kashino, S S; Singer-Vermes, L M; Calich, V L; Burger, E

    1990-09-01

    The in vitro subcultivation of some microorganisms for long periods causes measurable loss of their pathogenicity, which can be reverted by reisolation from infected hosts. We compared the pathogenicity and the in vitro growth pattern of one P. brasiliensis isolate (Pb 18) in its yeast phase, using the following samples: 1) The original pathogenic Pb 18 (OP). 2) Pb 18 attenuated by continuous in vitro subcultivation (AT). 3) Pb 18 (AT) reisolated from susceptible B10.A mice (RS). 4) Pb 18 (AT) reisolated from resistant A/SN mice (RR). Pathogenicity was evaluated by anatomopathology and mortality of mice infected i.p. with 5 x 10(6) fungi. Median survival times of mice infected with OP ranged from 74 to 117 days during the first 51 months of subculturing; with more cycles of subculturing the median survival time increased, reaching 250 days at the 64th month. This indicated decreasing virulence of OP during this period of subculturing. Survival of mice infected with RS and RR was respectively 112 and 123 days, which is similar to the behavior of the OP variant. The in vitro growth curve profile of RR showed significantly higher numbers of total and viable yeasts than the other studied variant. These results show that: 1) Pb 18 isolate loses its pathogenicity by continuous subcultivation. This phenomenon is reverted by reisolation from mice, independently from their susceptibility to the fungus; 2) the in vitro growth patterns of Pb 18 do not correlate with alterations in pathogenicity but are influenced by the host's environment. PMID:2233986

  2. Pathogenicity and immunogenicity of Paracoccidioides brasiliensis isolates in the human disease and in an experimental murine model.

    PubMed Central

    Singer-Vermes, L M; Burger, E; Calich, V L; Modesto-Xavier, L H; Sakamoto, T N; Sugizaki, M F; Meira, D A; Mendes, R P

    1994-01-01

    The pathogenicity and immunogenicity of six recently isolated Paracoccidioides brasiliensis samples derived from patients presenting distinct and well defined clinical forms of paracoccidioidomycosis (PCM) were compared as to their virulence, tropism to different organs and ability to induce specific cellular and humoral immune response in susceptible (B10.A) inbred mice. Isolates Pb44 and Pb47 were obtained from acute cases, Pb50 from a chronic severe form, Pb45 from a chronic moderate case and both Pb56 and Pb57 from chronic mild forms of PCM. Pathogenicity and tropism of each fungal sample were evaluated by LD50% estimation, examination of gross lesions on various organs at 2, 4, 12 and 16 weeks post-infection, and by colony-forming unit (CFU) counts in the lungs at week 16 post-infection of mice. Fungal tropism in human PCM and in B10.A mice was always dissociated. A well defined relationship between virulence of the fungal sample and the clinical findings of the correspondent patient was not evident, although a tendency to higher LD50% and less intense paracoccidioidic lesions was observed in mice infected with Pb56 and Pb57. The specific DTH response patterns varied according to the infectant sample, but positive DTH reactions at the beginning of the infection and a tendency to anergy or low DTH responses at week 12 and/or week 16 post-infection were always observed. A correspondence between the DTH response in humans and in mice was noticeable only when the isolates from the most benign cases (Pb56 and Pb57) were considered. The specific antibody patterns in mice and in the correspondent patients were also not analogous. Collectively, these results indicate that an association between the fungal pathogenicity and immunogenicity in the human disease and in susceptible mice was discernible only when isolates obtained from very mild cases (Pb56 and Pb57) were considered. PMID:8033408

  3. Low-level laser therapy to the mouse femur enhances the fungicidal response of neutrophils against Paracoccidioides brasiliensis.

    PubMed

    Burger, Eva; Mendes, Ana Carolina S C; Bani, Giulia M A C; Brigagão, Maísa R P L; Santos, Gérsika B; Malaquias, Luiz Cosme C; Chavasco, Jorge Kleber; Verinaud, Liana M; de Camargo, Zoilo P; Hamblin, Michael R; Sperandio, Felipe F

    2015-02-01

    Neutrophils (PMN) play a central role in host defense against the neglected fungal infection paracoccidioidomycosis (PCM), which is caused by the dimorphic fungus Paracoccidioides brasiliensis (Pb). PCM is of major importance, especially in Latin America, and its treatment relies on the use of antifungal drugs. However, the course of treatment is lengthy, leading to side effects and even development of fungal resistance. The goal of the study was to use low-level laser therapy (LLLT) to stimulate PMN to fight Pb in vivo. Swiss mice with subcutaneous air pouches were inoculated with a virulent strain of Pb or fungal cell wall components (Zymosan), and then received LLLT (780 nm; 50 mW; 12.5 J/cm2; 30 seconds per point, giving a total energy of 0.5 J per point) on alternate days at two points on each hind leg. The aim was to reach the bone marrow in the femur with light. Non-irradiated animals were used as controls. The number and viability of the PMN that migrated to the inoculation site was assessed, as well as their ability to synthesize proteins, produce reactive oxygen species (ROS) and their fungicidal activity. The highly pure PMN populations obtained after 10 days of infection were also subsequently cultured in the presence of Pb for trials of protein production, evaluation of mitochondrial activity, ROS production and quantification of viable fungi growth. PMN from mice that received LLLT were more active metabolically, had higher fungicidal activity against Pb in vivo and also in vitro. The kinetics of neutrophil protein production also correlated with a more activated state. LLLT may be a safe and non-invasive approach to deal with PCM infection. PMID:25675431

  4. Acetylcholine and ATP are coreleased from the electromotor nerve terminals of Narcine brasiliensis by an exocytotic mechanism.

    PubMed

    Unsworth, C D; Johnson, R G

    1990-01-01

    Although the exocytotic mechanism for quantal acetylcholine (ACh) release has been widely accepted for many years, it has repeatedly been challenged by reports that ACh released upon stimulation originates from the cytosol rather than synaptic vesicles. In this report, two independent experimental approaches were taken to establish the source of ACh released from the electromotor system of Narcine brasiliensis. Since ATP is colocalized with ACh in the cholinergic vesicle, the exocytotic theory predicts the corelease of these two components with a stoichiometry identical to that of the vesicle contents. The stimulated release of ATP from isolated synaptosomes could be accurately quantitated in the presence of the ATPase inhibitor adenosine 5'-[alpha, beta-methylene]triphosphate (500 microM), which prevented degradation of the released ATP. Various concentrations of elevated extracellular potassium (25-75 mM), veratridine (100 microM), and the calcium ionophore ionomycin (5 microM) all induced the corelease of ACh and ATP in a constant molar ratio of 5-6:1 (ACh/ATP), a stoichiometry consistent with that established for the vesicle content. In parallel to these stoichiometry studies, the compound 2-(4-phenylpiperidino)cyclohexanol (AH5183) was used to inhibit specifically the vesicular accumulation of newly synthesized (radiolabeled) ACh without affecting cytosolic levels of newly synthesized ACh in cholinergic nerve terminals. Treatment with AH5183 (10 microM) was shown to inhibit the release of newly synthesized ACh without markedly affecting total ACh release; thus, the entry of newly synthesized ACh into the synaptic vesicle is essential for its release. We conclude that ACh released upon stimulation originates exclusively from the vesicular pool and is coreleased stoichiometrically with other soluble vesicle contents. PMID:2137245

  5. The roles of quantum correlations in quantum cloning

    NASA Astrophysics Data System (ADS)

    Zhang, Jun; xiong Wu, Shao-; Yu, Chang-shui

    2014-12-01

    In this paper, we study the entanglement and quantum discord of the output modes in the unified 1 → 2 state-dependent cloning and probabilistic quantum cloning. The tripartite entanglement among the output modes and the quantum cloning machine is also considered. We find that the roles of the quantum correlations including the bipartite and tripartite entanglement and quantum discord strongly depend on the quantum cloning machines as well as the cloned state. In particular, it is found that this quantum cloning scheme can be realizable even without any quantum correlation.

  6. Whole genome comparison of donor and cloned dogs

    PubMed Central

    Kim, Hak-Min; Cho, Yun Sung; Kim, Hyunmin; Jho, Sungwoong; Son, Bongjun; Choi, Joung Yoon; Kim, Sangsoo; Lee, Byeong Chun; Bhak, Jong; Jang, Goo

    2013-01-01

    Cloning is a process that produces genetically identical organisms. However, the genomic degree of genetic resemblance in clones needs to be determined. In this report, the genomes of a cloned dog and its donor were compared. Compared with a human monozygotic twin, the genome of the cloned dog showed little difference from the genome of the nuclear donor dog in terms of single nucleotide variations, chromosomal instability, and telomere lengths. These findings suggest that cloning by somatic cell nuclear transfer produced an almost identical genome. The whole genome sequence data of donor and cloned dogs can provide a resource for further investigations on epigenetic contributions in phenotypic differences. PMID:24141358

  7. Changes in the gut microbiota of cloned and non-cloned control pigs during development of obesity: gut microbiota during development of obesity in cloned pigs

    PubMed Central

    2013-01-01

    Background Obesity induced by a high-caloric diet has previously been associated with changes in the gut microbiota in mice and in humans. In this study, pigs were cloned to minimize genetic and biological variation among the animals with the aim of developing a controlled metabolomic model suitable for a diet-intervention study. Cloning of pigs may be an attractive way to reduce genetic influences when investigating the effect of diet and obesity on different physiological sites. The aim of this study was to assess and compare the changes in the composition of the gut microbiota of cloned vs. non-cloned pigs during development of obesity by a high-fat/high-caloric diet. Furthermore, we investigated the association between diet-induced obesity and the relative abundance of the phyla Firmicutes and Bacteroidetes in the fecal-microbiota. The fecal microbiota from obese cloned (n = 5) and non-cloned control pigs (n= 6) was investigated biweekly over a period of 136 days, by terminal restriction fragment length polymorphism (T-RFLP) and quantitative real time PCR (qPCR). Results A positive correlation was observed between body-weight at endpoint and percent body-fat in cloned (r=0.9, P<0.0001) and in non-cloned control pigs (r=0.9, P<0.0001). Shannon Weaver and principal component analysis (PCA) of the terminal restriction fragments (T-RFs) revealed no differences in the bacterial composition or variability of the fecal microbiota between the cloned pigs or between cloned and non-cloned control pigs. Body-weight correlated positively with the relative abundance of Firmicutes in both cloned (r=0.37; P<0.02) and non cloned-control pigs (r=0.45; P<0.006), and negatively with the abundance of Bacteroidetes in cloned pigs (r=−0.33, P<0.04), but not in the non-cloned control pigs. Conclusion The cloned pigs did not have reduced inter-individual variation as compared to non-cloned pigs in regard to their gut microbiota in neither the obese nor the lean state. Diet

  8. Comparing quantum cloning: A Fisher-information perspective

    NASA Astrophysics Data System (ADS)

    Song, Hongting; Luo, Shunlong; Li, Nan; Chang, Lina

    2013-10-01

    Perfect cloning of an unknown quantum state is impossible. Approximate cloning, which is optimal in various senses, has been found in many cases. Paradigmatic examples are Wootters-Zurek cloning and universal cloning. These cloning machines aim at optimal cloning of the full quantum states. However, in practice, what is important and relevant may only involve partial information in quantum states, rather than quantum states themselves. For example, signals are often encoded as parameters in quantum states, whose information content is well synthesized by quantum Fisher information. This raises the basic issue of evaluating the information transferring capability (e.g., distributing quantum Fisher information) of quantum cloning. We assess and compare Wootters-Zurek cloning and universal cloning from this perspective and show that, on average, Wootters-Zurek cloning performs better than universal cloning for the phase (as well as amplitude) parameter, although they are incomparable individually, and universal cloning has many advantages over Wootters-Zurek cloning in other contexts. Physical insights and related issues are further discussed.

  9. Genetic Crossing vs Cloning by Computer Simulation

    NASA Astrophysics Data System (ADS)

    Dasgupta, Subinay

    We perform Monte Carlo simulation using Penna's bit string model, and compare the process of asexual reproduction by cloning with that by genetic crossover. We find them to be comparable as regards survival of a species, and also if a natural disaster is simulated.

  10. Genetic crossing vs cloning by computer simulation

    SciTech Connect

    Dasgupta, S.

    1997-06-01

    We perform Monte Carlo simulation using Penna`s bit string model, and compare the process of asexual reproduction by cloning with that by genetic crossover. We find them to be comparable as regards survival of a species, and also if a natural disaster is simulated.

  11. [Placental developmental defects in cloned mammalian animals].

    PubMed

    Ao, Zheng; Liu, Dewu; Cai, Gengyuan; Wu, Zhenfang; Li, Zicong

    2016-05-01

    The cloning technique, also called somatic cell nuclear transfer (SCNT), has been successfully established and gradually applied to various mammalian species. However, the developmental rate of SCNT mammalian embryos is very low, usually at 1% to 5%, which limits the application of SCNT. Placental developmental defects are considered as the main cause of SCNT embryo development inhibition. Almost all of SCNT-derived mammalian placentas exhibit various abnormalities, such as placental hyperplasia, vascular defects and umbilical cord malformation. Mechanistically, these abnormalities result from failure of establishment of correct epigenetic modification in the trophectoderm genome, which leads to erroneous expression of important genes for placenta development-related, particularly imprinted genes. Consequently, aberrant imprinted gene expression gives rise to placental morphologic abnormalities and functional defects, therefore decreases developmental competence of cloned embryos. Currently, although numerous methods that can improve the developmental ability of SCNT-derived embryos have been reported, most of them are unable to substantially enhance the success rate of SCNT due to failure to eliminate the placental development defects. In this review, we summarize placental abnormalities and imprinted gene expression in mammalian cloning, and propose directions for the future research aiming to improve the cloning efficiency. PMID:27232488

  12. Cloning: Learning to Replay the Genetic Tape.

    ERIC Educational Resources Information Center

    Holden, David J.

    1979-01-01

    Describes how plants can be produced by cloning by using tissue culture methods to mass-produce rare native prairie plants and trying to transfer some of the genetic characteristics of native grasses into cultivated cereals. The experiment was conducted at South Dakota State University. (HM)

  13. Computerized Adaptive Testing with Item Cloning.

    ERIC Educational Resources Information Center

    Glas, Cees A. W.; van der Linden, Wim J.

    2003-01-01

    Developed a multilevel item response (IRT) model that allows for differences between the distributions of item parameters of families of item clones. Results from simulation studies based on an item pool from the Law School Admission Test illustrate the accuracy of the item pool calibration and adaptive testing procedures based on the model. (SLD)

  14. Increasing efficiency in production of cloned piglets.

    PubMed

    Callesen, Henrik; Liu, Ying; Pedersen, Hanne S; Li, Rong; Schmidt, Mette

    2014-12-01

    The low efficiency in obtaining piglets after production of cloned embryos was challenged in two steps-first by performing in vitro culture for 5-6 days after cloning to obtain later-stage embryos for more precise selection for transfer, and second by reducing the number of embryos transferred per recipient sow. The data set consisted of combined results from a 4-year period where cloning was performed to produce piglets that were transgenic for important human diseases. For this, different transgenes and cell types were used, and the cloning work was performed by several persons using oocytes from different pig breeds, but following a standardized and optimized protocol. Results showed that in vitro culture is possible with a relatively stable rate of transferable embryos around 41% and a pregnancy rate around 90%. Furthermore, a reduction from around 80 embryos to 40 embryos transferred per recipient was possible without changing the efficiency of around 14% (piglets born out of embryos transferred). It was concluded that this approach can increase the efficiency in obtaining piglets by means of in vitro culture and selection of high-quality embryos with subsequent transfer into more recipients. Such changes can also reduce the need for personnel, time, and material when working with this technology. PMID:25333333

  15. Introduction to cloning by nuclear transplantation.

    PubMed

    Galli, Cesare; Lagutina, Irina; Lazzari, Giovanna

    2003-01-01

    Despite its long history, the cloning of animals by nuclear transplantation is going through a "renaissance" after the birth of Dolly. The amount of work and achievements obtained in the last seven years are probably greater than those obtained in half a century of research. However, the principal obstacles outlined years ago with the work on somatic cell cloning in amphybia, are all still there in mammals. The importance of somatic cell nuclear transfer is, without any doubt, beyond the scope of replicating superior animal genotypes. It is an invaluable experimental tool to address fundamental scientific issues such as nuclear potency, cell de-differentiation, chromatin structure and function, epigenetics, and genome manipulation. For these reasons the importance of cloning is not for what it can achieve but for the technical support it can provide to biomedical research and in particular to the study of epigenetics, cancer and stem cell biology, cell therapy and regenerative medicine. In this introductory paper we will summarize the intellectual and technical framework of cloning animals by nuclear transfer that still remains the only absolute way of judging the success of the procedure. Together with the achievements of the recent past we will mention the very last developments and the many questions that still remain open. Current research efforts are expected to provide some answers and certainly new questions. PMID:14733742

  16. Molecular Individual-Based Approach on Triatoma brasiliensis: Inferences on Triatomine Foci, Trypanosoma cruzi Natural Infection Prevalence, Parasite Diversity and Feeding Sources

    PubMed Central

    Almeida, Carlos Eduardo; Faucher, Leslie; Lavina, Morgane; Costa, Jane; Harry, Myriam

    2016-01-01

    We used an individual-based molecular multisource approach to assess the epidemiological importance of Triatoma brasiliensis collected in distinct sites and ecotopes in Rio Grande do Norte State, Brazil. In the semi-arid zones of Brazil, this blood sucking bug is the most important vector of Trypanosoma cruzi—the parasite that causes Chagas disease. First, cytochrome b (cytb) and microsatellite markers were used for inferences on the genetic structure of five populations (108 bugs). Second, we determined the natural T. cruzi infection prevalence and parasite diversity in 126 bugs by amplifying a mini-exon gene from triatomine gut contents. Third, we identified the natural feeding sources of 60 T. brasiliensis by using the blood meal content via vertebrate cytb analysis. Demographic inferences based on cytb variation indicated expansion events in some sylvatic and domiciliary populations. Microsatellite results indicated gene flow between sylvatic and anthropic (domiciliary and peridomiciliary) populations, which threatens vector control efforts because sylvatic population are uncontrollable. A high natural T. cruzi infection prevalence (52–71%) and two parasite lineages were found for the sylvatic foci, in which 68% of bugs had fed on Kerodon rupestris (Rodentia: Caviidae), highlighting it as a potential reservoir. For peridomiciliary bugs, Galea spixii (Rodentia: Caviidae) was the main mammal feeding source, which may reinforce previous concerns about the potential of this animal to link the sylvatic and domiciliary T. cruzi cycles. PMID:26891047

  17. Feeding habits of the congeneric species Stellifer rastrifer and Stellifer brasiliensis (Acanthopterygii: Sciaenidae) co-occurring in the coast of the state of Santa Catarina, Brazil.

    PubMed

    Sabinson, L M; Rodrigues-Filho, J L; Peret, A C; Branco, J O; Verani, J R

    2015-05-01

    The objective of the present study was to investigate the feeding habits allowing the occurrence of the congeneric species Stellifer rastrifer and Stellifer brasiensis, collected on a seasonal basis, in the operating area of the artisanal seabob-shrimp trawl fishery in Porto Belo, an important fishing area on the southern Brazilian coast. Fish were collected seasonally from November 2009 to August 2010, in isobaths of 10, 20 and 30 m. Of the 450 stomachs analyzed, 194 belonged to Stellifer rastrifer and 242 to Stellifer brasiliensis. From the totality of analyzed stomachs 33.49% were empty and 66.52% had some content. Crustacea were the predominant food category in the diet of S. rastrifer and S. brasiliensis, and the shrimp Acetes americanus was the most frequent crustacean in stomachs of both studied species. The overlapping in the diet of the species was high, however when analyzing the overlap in each season it was found that it occurred only during spring and winter. Although the most important items in the diet of these species are the same, the intake of each alternates in time, suggesting a difference in predation pressure, which leads to less direct competition in a given time. PMID:26132027

  18. Transcription levels of CHS5 and CHS4 genes in Paracoccidioides brasiliensis mycelial phase, respond to alterations in external osmolarity, oxidative stress and glucose concentration.

    PubMed

    Niño-Vega, Gustavo A; Sorais, Françoise; San-Blas, Gioconda

    2009-10-01

    The complete sequence of Paracoccidioides brasiliensis CHS5 gene, encoding a putative chitin synthase revealed a 5583nt open reading frame, interrupted by three introns of 82, 87 and 97bp (GenBank Accession No EF654132). The deduced protein contains 1861 amino acids with a predicted molecular weight of 206.9kDa. Both its large size and the presence of a N-terminal region of approx. 800 residues with a characteristic putative myosin motor-like domain, allow us to include PbrChs5 into class V fungal chitin synthases. Sequence analysis of over 4kb from the 5' UTR region in CHS5, revealed the presence of a previously reported CHS4 gene in P. brasiliensis, arranged in a head-to-head configuration with CHS5. A motif search in this shared region showed the presence of stress response elements (STREs), three binding sites for the transcription activators Rlm1p (known to be stimulated by hypo-osmotic stress) and clusters of Adr1 (related to glucose repression). A quantitative RT-PCR analysis pointed to changes in transcription levels for both genes following oxidative stress, alteration of external osmolarity and under glucose-repressible conditions, suggesting a common regulatory mechanism of transcription. PMID:19616626

  19. Babesia bovis clones: biochemical and enzymatic characterization

    SciTech Connect

    Rodriguez Camarillo, S.D.

    1985-01-01

    Studies were undertaken to generate additional knowledge of the biochemistry of Babesia bovis. A modified in vitro culture technique used for cloning B. bovis. This technique included a low oxygen concentration atmosphere (2%, O/sub 2/, 5% CO/sub 2/, 93% N/sub 2/) and 4 mm fluid level. Cultures initiated with one infected erythrocyte were maintained until parasitemias of positive wells reached 2% parasitemia. Primary clones were obtained and from these, nine clones were recloned twice and used for subsequent studies. A procedure was developed to concentrate and separate B. bovis merozoites and infected erythrocytes by Percoll density gradients. Merozoites separated at 1.087 g/ml specific density, whereas infected erythrocytes separated at 1.121 g/ml. Viability of purified parasites was not affected. Agarose gel electrophoresis was used to identify metabolic enzyme in B. bovis and B. bigemina. The enzymes LDH, GDH, GPI and HK were detected in both species. Molecular analysis by one and two-dimensional gel electrophoresis of proteins metabolically labeled with /sup 35/S-methionine indicated that two clones, derived from the same field strain, were similar but not identical to the parent. Fewer proteins were observed in the parental strain. Growth of two 60-Co irradiated B. bovis clones indicated a dose-effect relationship. Growth of parasites exposed for the longest period was initially retarded but returned to normal growth after two or three subcultures. Cultures exposed for shorter periods were unaffected with respect to the rate of growth. Analysis of electrophoretic mobility of metabolic enzyme showed a change in migration pattern.

  20. IVA cloning: A single-tube universal cloning system exploiting bacterial In Vivo Assembly

    PubMed Central

    García-Nafría, Javier; Watson, Jake F.; Greger, Ingo H.

    2016-01-01

    In vivo homologous recombination holds the potential for optimal molecular cloning, however, current strategies require specialised bacterial strains or laborious protocols. Here, we exploit a recA-independent recombination pathway, present in widespread laboratory E.coli strains, to develop IVA (In Vivo Assembly) cloning. This system eliminates the need for enzymatic assembly and reduces all molecular cloning procedures to a single-tube, single-step PCR, performed in <2 hours from setup to transformation. Unlike other methods, IVA is a complete system, and offers significant advantages over alternative methods for all cloning procedures (insertions, deletions, site-directed mutagenesis and sub-cloning). Significantly, IVA allows unprecedented simplification of complex cloning procedures: five simultaneous modifications of any kind, multi-fragment assembly and library construction are performed in approximately half the time of current protocols, still in a single-step fashion. This system is efficient, seamless and sequence-independent, and requires no special kits, enzymes or proprietary bacteria, which will allow its immediate adoption by the academic and industrial molecular biology community. PMID:27264908