WorldWideScience

Sample records for involving selective isolation

  1. Selective factors involved in oil flotation isolation of black yeasts from the environment

    NARCIS (Netherlands)

    Satow, M.M.; Attili-Angelis, D.; de Hoog, G.S.; Angelis, D.F.; Vicente, V.A.

    2008-01-01

    The oil flotation isolation technique has been successfully applied to recover chaetothyrialean black yeasts and relatives from the environment. The selective mechanisms playing a role in isolation are unknown. The fungi concerned are supposed to occupy specialized microniches in nature, taking adva

  2. Selective factors involved in oil flotation isolation of black yeasts from the environment

    OpenAIRE

    Satow, M.M.; Attili-Angelis, D. [UNESP; de Hoog, G. S.; Angelis, D. F. [UNESP; Vicente, V.A.

    2008-01-01

    The oil flotation isolation technique has been successfully applied to recover chaetothyrialean black yeasts and relatives from the environment. The selective mechanisms playing a role in isolation are unknown. The fungi concerned are supposed to occupy specialized microniches in nature, taking advantage of (1) oligotrophism. Mineral oil as a main selective agent may be based on (2) hydrophobicity or on (3) assimilation. All three hypotheses are tested in this paper. Results show that cell wa...

  3. Selective factors involved in oil flotation isolation of black yeasts from the environment.

    Science.gov (United States)

    Satow, M M; Attili-Angelis, D; de Hoog, G S; Angelis, D F; Vicente, V A

    2008-01-01

    The oil flotation isolation technique has been successfully applied to recover chaetothyrialean black yeasts and relatives from the environment. The selective mechanisms playing a role in isolation are unknown. The fungi concerned are supposed to occupy specialized microniches in nature, taking advantage of (1) oligotrophism. Mineral oil as a main selective agent may be based on (2) hydrophobicity or on (3) assimilation. All three hypotheses are tested in this paper. Results show that cell wall hydrophobicity is unlikely to be a selective factor. Incubation under poor nutrient conditions provides competitive advantage for black yeasts, especially for Exophiala strains, which are subsequently enriched by mineral oil which enhances growth in this group of fungi. Incubation under mineral media and mineral oil can be used as selective factor.

  4. Isolated lingual involvement in Wilson's disease.

    Science.gov (United States)

    Choudhary, Neera; Joshi, Laxmikant; Duggal, Ashish; Puri, Vinod; Khwaja, Geeta Anjum

    2015-01-01

    Lingual involvement can occur in a variety of neurological disorders including pyramidal, extrapyramidal and lower motor neuron disorders. It can be seen in the form of tremor, bradykinesia, dystonia, atrophy and weakness of tongue movements and can clinically present as difficulty in swallowing and dysarthria which can be a source of great discomfort to the patient. We describe a patient who presented with isolated lingual involvement and was diagnosed to have Wilsons's disease. This case emphasizes the clinical variability in presentation of Wilson's disease and importance of early clinical diagnosis.

  5. Selection and characterisation of monepantel resistance in Teladorsagia circumcincta isolates

    Directory of Open Access Journals (Sweden)

    D.J. Bartley

    2015-08-01

    To address these questions three MPTL-resistant Teladorsagia circumcincta isolates (MTci2-11, MTci5-13 and MTci7-12 have been artificially selected in vivo from phenotypically characterised parent isolates (MTci2, MTci5, MTci7 respectively. The selection process involved collecting and culturing eggs from surviving worms from sheep administered sub-optimal dosages of MPTL (Zolvix® to provide infective larvae to infect further sheep until resistant isolates were generated (between 9 and 13 rounds of selection. A controlled efficacy test was conducted using the original parental isolates and the newly generated MPTL resistant isolates (n = 5 per group. Selected isolates were assessed both under anthelmintic stress (Zolvix®, 2.5 mg/kg bodyweight; MTci-MPTL and at rest (untreated, MTci-CON. A number of life-history traits were assessed, namely, worm establishment rates, time to patency, faecal egg output, body length of adults and eggs in utero. The estimated resistance status of the selected isolates was confirmed with 48%, 28% and 9% reductions in worm burden at 7-days post Zolvix® administration for MTci2-11-MPTL, MTci5-13-MPTL and MTci7-12-MPTL, respectively, compared with untreated controls. One of the selected isolates MTci7-12-CON showed significantly greater total worm burden (p = 0.025, greater establishment rate (p = 0.033, decreased time to patency (p = 0.048, higher cumulative egg outputs (p = 0.002 compared with its parental derivative MTci7. The trial results suggest that anthelmintic selection in T. circumcincta, albeit under experimental conditions, can select for more prolific/fecund and quicker maturing populations. These data provide an insight into how parasites evolve in response to anthelmintic pressure.

  6. Isolated neurological involvement of lymphomatoid granulomatosis

    Institute of Scientific and Technical Information of China (English)

    HU Yu-hua; SHAO En-de; WU Jian-liang; MENG Xian-bing

    2010-01-01

    @@ Lymphomatoid granulomatosis (LG) is a rare EpsteinBarr virus (EBV)-associated systemic granulomatous disease that is characterized by an angiocentric and occasionally angiodestructive polymorphic cellular infiltrate. It most frequently affects the lungs, kidneys,and brain.1,2 Usually the lesions are multi-systemic and multifocal. Central nervous system (CNS) involvement occurs in approximately 30% of affected patients, and patients may present with nonspecific neurological symptoms.3 Although CNS lesions usually appear as a secondary metastasis of pulmonary lesions, in rare conditions, it may be the initial or only manifestation of the disease. In this report, we describe two cases presenting with unifocal mass in the brain, histologically confirmed to be LG,

  7. SELECTIVE EXTRACTION OF ISOLATED MITOTIC APPARATUS

    Science.gov (United States)

    Bibring, Thomas; Baxandall, Jane

    1971-01-01

    Mitotic apparatus isolated from sea urchin eggs has been treated with meralluride sodium under conditions otherwise resembling those of its isolation. The treatment causes a selective morphological disappearance of microtubules while extracting a major protein fraction, probably consisting of two closely related proteins, which constitutes about 10% of mitotic apparatus protein. Extraction of other cell particulates under similar conditions yields much less of this protein. The extracted protein closely resembles outer doublet microtubule protein from sea urchin sperm tail in properties considered typical of microtubule proteins: precipitation by calcium ion and vinblastine, electrophoretic mobility in both acid and basic polyacrylamide gels, sedimentation coefficient, molecular weight, and, according to a preliminary determination, amino acid composition. An antiserum against a preparation of sperm tail outer doublet microtubules cross-reacts with the extract from mitotic apparatus. On the basis of these findings it appears that microtubule protein is selectively extracted from isolated mitotic apparatus by treatment with meralluride, and is a typical microtubule protein. PMID:5543404

  8. Isolated lingual involvement in Wilson′s disease

    Directory of Open Access Journals (Sweden)

    Neera Choudhary

    2015-01-01

    Full Text Available Lingual involvement can occur in a variety of neurological disorders including pyramidal, extrapyramidal and lower motor neuron disorders. It can be seen in the form of tremor, bradykinesia, dystonia, atrophy and weakness of tongue movements and can clinically present as difficulty in swallowing and dysarthria which can be a source of great discomfort to the patient. We describe a patient who presented with isolated lingual involvement and was diagnosed to have Wilsons′s disease. This case emphasizes the clinical variability in presentation of Wilson′s disease and importance of early clinical diagnosis.

  9. Selected aspects of isolation of physically disabled people

    Directory of Open Access Journals (Sweden)

    Piotr Winczewski

    2014-04-01

    Full Text Available The presented material is a concise report of research carried out on the conditions of isolation of people with disabilities. This article highlights the influence of the selected factors on disabled people remaining isolated or escaping from solitude. The collected research material consists of biographical studies of selected cases.

  10. Visual pathways involvement in clinically isolated syndrome in children

    Institute of Scientific and Technical Information of China (English)

    Vladislav; Voitenkov; Natalia; Skripchenko; Andrey; Klimkin

    2015-01-01

    AIM: To investigate extent and nature of visual pathways involvement in children with clinically isolated syndrome(CIS).METHODS: Forty-seven patients(age 11-17y) with CIS, which later proved to be multiple sclerosis(MS)onset, and 30 controls underwent visual evoked potentials(VEP) investigation within 12 d from the appearance of the first signs of disease. Latency and amplitude of P100 peak were compared with normative data and between groups.RESULTS: In 58% patients, including those without signs of retrobulbar neuritis, significant slowing of conduction along the central visual pathways(P100latency lengthening) is seen. P100 amplitudes drop(signs of axonal damage) are registered less frequently(29% cases).CONCLUSION: The results indicate that visual pathways are often affected in the MS onset; mostly demyelination signs are seen. Despite MRI significance for MS diagnostic, VEPs proved to be still effective in early diagnosis of MS in children.

  11. Internalization of isolated functional mitochondria: involvement of macropinocytosis

    Science.gov (United States)

    Kitani, Tomoya; Kami, Daisuke; Matoba, Satoaki; Gojo, Satoshi

    2014-01-01

    In eukaryotic cells, mitochondrial dysfunction is associated with a variety of human diseases. Delivery of exogenous functional mitochondria into damaged cells has been proposed as a mechanism of cell transplant and physiological repair for damaged tissue. We here demonstrated that isolated mitochondria can be transferred into homogeneic and xenogeneic cells by simple co-incubation using genetically labelled mitochondria, and elucidated the mechanism and the effect of direct mitochondrial transfer. Intracellular localization of exogenous mitochondria was confirmed by PCR, real-time PCR, live fluorescence imaging, three-dimensional reconstruction imaging, continuous time-lapse microscopic observation, flow cytometric analysis and immunoelectron microscopy. Isolated homogeneic mitochondria were transferred into human uterine endometrial gland-derived mesenchymal cells in a dose-dependent manner. Moreover, mitochondrial transfer rescued the mitochondrial respiratory function and improved the cellular viability in mitochondrial DNA-depleted cells and these effects lasted several days. Finally, we discovered that mitochondrial internalization involves macropinocytosis. In conclusion, these data support direct transfer of exogenous mitochondria as a promising approach for the treatment of various diseases. PMID:24912369

  12. Hypertensive Encephalopathy: Isolated Pons Involvement Mimicking Central Pontine Myelinolysis

    Energy Technology Data Exchange (ETDEWEB)

    Gamanagatti, S.; Subramanian, S. [India Institute of Medical Sciences, New Delhi (India)

    2006-09-15

    MRI of the brain was performed, and it demonstrated an isolated high signal on the T2 weighted and fluid attenuated inversion recovery sequences that involved only the central pons with sparing the periphery. There was no restricted diffusion on diffusion weighted imaging. The differential diagnosis included posterior reversible syndrome and central pontine myelinolysis; however, the blood sodium on admission was normal. The pathogenesis of HE is that the auto-regulatory mechanisms that control the cerebral blood flow are exceeded, resulting in hyper-perfusion. The consequent over-distension of the cerebral vessels, the breakdown of the blood brain barrier and ultimately, the extravasation of fluid into the interstitium all cause vasogenic edema. In most cases, the changes of hypertensive encephalopathy represent reversible vasogenic edema, which can be seen on T2-weighted images, and restricted diffusion is not seen on the diffusion-weighted imaging (DWI) and the apparent diffusion coefficient (ADC) maps. Hypertensive encephalopathy that manifests as a reversible increased signal isolated to the pons on T2-weighted images is extremely uncommon. The differential diagnosis for such pontine T2 hyperintensity includes pontine glioma, ischemic and radiation changes (generally irreversible conditions), as well as central pontine myelinolysis (CPM) and demyelinating disorders such as multiple sclerosis, acute disseminated encephalomyelitis and rhomb-encephalitis. In CPM electrolyte imbalances provide a clue for the diagnosis, where as for glioma, there will be an expansion and mass effect. In conclusion, clinical recognition of brainstem HE may be difficult. The features of a lack of correlation between the severity of the radiological abnormality and the clinical status, combined with the rapid resolution following antihypertensive treatment, should suggest the diagnosis. It is important for the radiologist to be familiar with the imaging abnormalities of this life

  13. A wavelength selective bidirectional isolator for access optical networks

    Science.gov (United States)

    Hu, Xi-kui; Huang, Zhang-di; Li, Su-shan; Xu, Fei; Chen, Beckham; Lu, Yan-qing

    2011-05-01

    A wavelength selective bidirectional optical isolator is proposed. Being different from conventional isolators, a well-designed wave plate is employed and works together with the Faraday rotator. Different wavelengths thus experience different phase retardation so that wavelength-dependent polarization states are obtained for bidirectional beams. As an example, a (1.49 μm, 1.31 μm) wavelength selective isolator is proposed, which means only 1.49 μm light can propagate along one-direction while the opposite wave is just for 1.31 μm light. Over 60 dB optical isolation is obtained by selecting suitable wave plate thickness and orientation. This interesting isolator may have promising applications in access optical networks.

  14. Use of bacteriophage for the selective isolation of thermophilic actinomycetes from composted eucalyptus bark.

    Science.gov (United States)

    Kurtböke, D I; Murphy, N E; Sivasithamparam, K

    1993-01-01

    A method was developed to reduce the numbers of thermophilic bacteria on isolation plates, which in turn facilitated the detection and isolation of thermophilic actinomycetes. The method involves exposing the test material to bacteriophage suspensions prior to inoculation on isolation plates. This method was applied to composted eucalyptus bark samples, which were then inoculated on R8 and 1/2 TSA + 0.2% casein hydrolysate agar plates. The phage susceptibility of thermophilic bacteria provided a selective means of reducing their numbers on isolation plates and hence increased the numbers of Thermomonospora, Saccharopolyspora rectivirgula, and thermophilic Streptomyces spp. on these media in comparison with the numbers recorded from control plates.

  15. Patagonian red wines: selection of Lactobacillus plantarum isolates as potential starter cultures for malolactic fermentation.

    Science.gov (United States)

    Bravo-Ferrada, Bárbara Mercedes; Hollmann, Axel; Delfederico, Lucrecia; Valdés La Hens, Danay; Caballero, Adriana; Semorile, Liliana

    2013-09-01

    The aim of this study was to evaluate fifty-three Lactobacillus plantarum isolates obtained from a Patagonian red wine, molecularly identified and typified using RAPD analysis, in order to select starter cultures for malolactic fermentation (MLF). The results obtained suggest a considerable genetic diversity, taking into account that all L. plantarum isolates were obtained from one cellar and one vintage. Based on the capacity to tolerate a concentration of 14 % ethanol in MRS broth for 2 days, eight isolates were selected for the subsequent analysis. The incidence of various wine stress factors (ethanol, acid pH, lysozyme and sulfur dioxide) on isolates growth was studied. Besides, glucosidase and tannase activities were evaluated, and the presence of genes involved in the synthesis of biogenic amines was examined by PCR. A previously characterized indigenous Oenococcus oeni strain was included with comparative purposes. Differences in technologically relevant characteristics were observed among the eight L. plantarum selected isolates, revealing an isolate-dependent behavior. Detectable glucosidase and tannase activities were found in all isolates. The presence of genes encoding histidine and tyrosine descarboxylases and putrescine carbamoyltransferase was not detected. The ability of L. plantarum isolates to grow and consume L-malic acid in simulated laboratory-scale vinifications revealed that two of them could be considered as possible MLF starter cultures for Patagonian red wines. These isolates will be subjected to further analysis, for a final winery technological characterization.

  16. Isolation of microorganisms involved in reduction of crystalline iron(III oxides in natural environments

    Directory of Open Access Journals (Sweden)

    Tomoyuki eHori

    2015-05-01

    Full Text Available Reduction of crystalline Fe(III oxides is one of the most important electron sinks for organic compound oxidation in natural environments. Yet a limited number of isolates makes it difficult to understand physiology and ecological impact of the microorganisms involved. Here, two-staged cultivation was implemented to selectively enrich and isolate crystalline iron(III reducers in soils and sediments. Firstly, iron reducers were enriched and other untargeted eutrophs were depleted by two-year successive culture on a crystalline ferric iron oxide (i.e., goethite, lepidocrocite, hematite, or magnetite as electron acceptor. Fifty-eight out of 136 incubation conditions allowed the continued existence of microorganisms as confirmed by PCR amplification. High-throughput Illumina sequencing based on 16S rRNA genes revealed that the enrichment cultures on each of the ferric iron oxides contained bacteria belonging to the Deltaproteobacteria (mainly Geobacteraceae, followed by Firmicutes and Chloroflexi, which also comprised most of the operational taxonomic units (OTUs identified. The Venn diagrams indicated that the core OTUs enriched with all of the iron oxides were dominant in the Geobacteraceae. Secondly, 38 enrichment cultures including novel microorganisms were transferred to soluble-iron(III media in order to stimulate proliferation of the enriched iron reducers. Through extinction dilution-culture and single colony isolation, six strains within the Deltaproteobacteria were finally obtained; five strains belonged to the genus Geobacter and one strain to Pelobacter. These isolates had 94.8–98.1% sequence similarities of 16S rRNA genes to cultured relatives. All the isolates were able to grow on acetate and ferric iron but their physiological characteristics differed considerably in terms of growth rate. The results demonstrate the successful enrichment and isolation of novel iron(III reducers that were able to thrive by reducing highly

  17. Microalgae Isolation and Selection for Prospective Biodiesel Production

    Directory of Open Access Journals (Sweden)

    Peer M. Schenk

    2012-06-01

    Full Text Available Biodiesel production from microalgae is being widely developed at different scales as a potential source of renewable energy with both economic and environmental benefits. Although many microalgae species have been identified and isolated for lipid production, there is currently no consensus as to which species provide the highest productivity. Different species are expected to function best at different aquatic, geographical and climatic conditions. In addition, other value-added products are now being considered for commercial production which necessitates the selection of the most capable algae strains suitable for multiple-product algae biorefineries. Here we present and review practical issues of several simple and robust methods for microalgae isolation and selection for traits that maybe most relevant for commercial biodiesel production. A combination of conventional and modern techniques is likely to be the most efficient route from isolation to large-scale cultivation.

  18. Selective isolation of Yersinia pestis from plague-infected fleas

    Science.gov (United States)

    Sarovich, Derek S.; Colman, Rebecca E.; Price, Erin P.; Chung, Wai Kwan; Lee, Judy; Schupp, James M.; Alexander, James; Keim, Paul; Wagner., David M.

    2010-01-01

    We evaluated Yersinia CIN agar for the isolation of Yersinia pestis from infected fleas. CIN media is effective for the differentiation of Y. pestis from flea commensal flora and is sufficiently inhibitory to other bacteria that typically outcompete Y. pestis after 48 hours of growth using less selective media. PMID:20385178

  19. Characterization of root-nodule bacteria isolated from Vicia faba and selection of plant growth promoting isolates.

    Science.gov (United States)

    Saïdi, Sabrine; Chebil, Samir; Gtari, Maher; Mhamdi, Ridha

    2013-06-01

    A collection of 104 isolates from root-nodules of Vicia faba was submitted to 16S rRNA PCR-RFLP typing. A representative sample was further submitted to sequence analysis of 16S rRNA. Isolates were assigned to 12 genera. All the nodulating isolates (45 %) were closely related to Rhizobium leguminosarum USDA2370(T) (99.34 %). The remaining isolates, including potential human pathogens, failed to nodulate their original host. They were checked for presence of symbiotic genes, P-solubilization, phytohormone and siderophore production, and then tested for their growth promoting abilities. Results indicated that 9 strains could induce significant increase (41-71 %) in shoot dry yield of faba bean. A Pseudomonas strain was further assessed in on-farm trial in combination with a selected rhizobial strain. This work indicated that nodule-associated bacteria could be a valuable pool for selection of effective plant growth promoting isolates. Nevertheless, the possible involvement of nodules in increasing risks related to pathogenic bacteria should not be neglected and needs to be investigated further.

  20. Fault detection and isolation in processes involving induction machines

    Energy Technology Data Exchange (ETDEWEB)

    Zell, K.; Medvedev, A. [Control Engineering Group, Luleaa University of Technology, Luleaa (Sweden)

    1997-12-31

    A model-based technique for fault detection and isolation in electro-mechanical systems comprising induction machines is introduced. Two coupled state observers, one for the induction machine and another for the mechanical load, are used to detect and recognize fault-specific behaviors (fault signatures) from the real-time measurements of the rotor angular velocity and terminal voltages and currents. Practical applicability of the method is verified in full-scale experiments with a conveyor belt drive at SSAB, Luleaa Works. (orig.) 3 refs.

  1. [Takayasu's disease disclosed by isolated involvement of the ascending aorta].

    Science.gov (United States)

    Marcaggi, X; Courant, N; Soubrier, M; Kemeny, J L; Camilleri, L; Lusson, J R; Cassagnes, J

    1992-03-01

    The authors report the histological discovery of a case of Takayasu syndrome affecting the ascending aorta. This involvement appearing to concern only the aorta, with no symptomatic complaints nor any laboratory abnormalities indicative of an inflammatory syndrome, corticosteroids were not prescribed. Management consisted of biennial monitoring by transthoracic and transesophageal ultrasonography of the aorta and the supra-aortic main vessels together with monitoring of laboratory parameters.

  2. Involvement of chloride in renin secretion from isolated rat glomeruli

    DEFF Research Database (Denmark)

    Skøtt, O; Jensen, B L

    1992-01-01

    M) inhibited renin release reversibly. Substitution of Cl- with nitrate (101 mM) stimulated renin secretion. Substitution with iodide (15 or 101 mM) had no consistent effect. The stimulation induced by calcium-free solutions was high in May and low in September. In the absence of chloride, the response....... It is concluded that the renin secretory process has a demand for permeant anions. The stimulation caused by low external calcium involves at least two mechanisms: one is anion sensitive, powerful, varies with the season, and includes a recruitment phenomenon; another is anion insensitive and weak....

  3. Selection of Infective Arbuscular Mycorrhizal Fungal Isolates for Field Inoculation

    Directory of Open Access Journals (Sweden)

    Elisa Pellegrino

    2010-09-01

    Full Text Available Arbuscular mycorrhizal (AM fungi play a key role in host plant growth and health, nutrient and water uptake, plant community diversity and dynamics. AM fungi differ in their symbiotic performance, which is the result of the interaction of two fungal characters, infectivity and efficiency. Infectivity is the ability of a fungal isolate to establish rapidly an extensive mycorrhizal symbiosis and is correlated with pre-symbiotic steps of fungal life cycle, such as spore germination and hyphal growth. Here, different AM fungal isolates were tested, with the aim of selecting infective endophytes for field inoculation. Greenhouse and microcosm experiments were performed in order to assess the ability of 12 AM fungal isolates to produce spores, colonize host roots and to perform initial steps of symbiosis establishment, such as spore germination and hyphal growth. AM fungal spore production and root colonization were significantly different among AM fungal isolates. Spore and sporocarp densities ranged from 0.8 to 7.4 and from 0.6 to 2.0 per gram of soil, respectively, whereas root colonization ranged from 2.9 to 72.2%. Percentage of spore or sporocarp germination ranged from 5.8 to 53.3% and hyphal length from 4.7 to 79.8 mm. The ordination analysis (Redundancy Analysis, RDA showed that environmental factors explained about 60% of the whole variance and their effect on fungal infectivity variables was significant (P = 0.002. The biplot clearly showed that variables which might be used to detect infective AM fungal isolates were hyphal length and root colonization. Such analysis may allow the detection of the best parameters to select efficient AM fungal isolates to be used in agriculture.

  4. Selection, isolation, and identification of fungi for bioherbicide production.

    Science.gov (United States)

    Souza, Angélica Rossana Castro de; Baldoni, Daiana Bortoluzzi; Lima, Jessica; Porto, Vitória; Marcuz, Camila; Machado, Carolina; Ferraz, Rafael Camargo; Kuhn, Raquel C; Jacques, Rodrigo J S; Guedes, Jerson V C; Mazutti, Marcio A

    Production of a bioherbicide for biological control of weeds requires a series of steps, from selection of a suitable microbial strain to final formulation. Thus, this study aimed to select fungi for production of secondary metabolites with herbicidal activity using biological resources of the Brazilian Pampa biome. Phytopathogenic fungi were isolated from infected tissues of weeds in the Pampa biome. A liquid synthetic culture medium was used for production of metabolites. The phytotoxicity of fungal metabolites was assessed via biological tests using the plant Cucumis sativus L., and the most promising strain was identified by molecular analysis. Thirty-nine fungi were isolated, and 28 presented some phytotoxic symptoms against the target plant. Fungus VP51 belonging to the genus Diaporthe showed the most pronounced herbicidal activity. The Brazilian Pampa biome is a potential resource for the development of new and sustainable chemical compounds for modern agriculture. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  5. Selection, isolation, and identification of fungi for bioherbicide production

    Directory of Open Access Journals (Sweden)

    Angélica Rossana Castro de Souza

    Full Text Available Abstract Production of a bioherbicide for biological control of weeds requires a series of steps, from selection of a suitable microbial strain to final formulation. Thus, this study aimed to select fungi for production of secondary metabolites with herbicidal activity using biological resources of the Brazilian Pampa biome. Phytopathogenic fungi were isolated from infected tissues of weeds in the Pampa biome. A liquid synthetic culture medium was used for production of metabolites. The phytotoxicity of fungal metabolites was assessed via biological tests using the plant Cucumis sativus L., and the most promising strain was identified by molecular analysis. Thirty-nine fungi were isolated, and 28 presented some phytotoxic symptoms against the target plant. Fungus VP51 belonging to the genus Diaporthe showed the most pronounced herbicidal activity. The Brazilian Pampa biome is a potential resource for the development of new and sustainable chemical compounds for modern agriculture.

  6. Characterization of Staphylococcus caprae Clinical Isolates Involved in Human Bone and Joint Infections, Compared with Goat Mastitis Isolates.

    Science.gov (United States)

    d'Ersu, J; Aubin, G G; Mercier, P; Nicollet, P; Bémer, P; Corvec, S

    2016-01-01

    Staphylococcus caprae is an emerging microorganism in human bone and joint infections (BJI). The aim of this study is to describe the features of S. caprae isolates involved in BJI (H for human) compared with those of isolates recovered in goat mastitis (A for animal). Fourteen isolates of each origin were included. Identifications were performed using a Vitek 2 GP ID card, tuf gene sequencing, and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) Vitek MS. Molecular typing was carried out using pulsed-field gel electrophoresis (PFGE) and DiversiLab technology. The crystal violet method was used to determine biofilm-forming ability. Virulence factors were searched by PCR. Vitek MS technology provides an accurate identification for the two types of isolates compared to that of gold-standard sequencing (sensitivity, 96.4%), whereas the Vitek 2 GP ID card was more effective for H isolates. Molecular typing methods revealed two distinct lineages corresponding to the origin despite few overlaps: H and A. In our experimental conditions, no significant difference was observed in biofilm production ability between H and A isolates. Nine isolates (5 H isolates and 4 A isolates) behaved as weak producers while one A isolate was a strong producer. Concerning virulence factors, the autolysin atlC and the serine aspartate adhesin (sdrZ) genes were detected in 24 isolates (86%), whereas the lipase gene was always detected, except in one H isolate (96%). The ica operon was present in 23 isolates (82%). Fibrinogen-binding (fbe) or collagen-binding (cna) genes were not detected by using primers designed for Staphylococcus aureus or Staphylococcus epidermidis, even in low stringency conditions. Although S. caprae probably remains underestimated in human infections, further studies are needed to better understand the evolution and the adaptation of this species to its host.

  7. An Isolated Bee Sting Involving Multiple Cranial Nerves

    Directory of Open Access Journals (Sweden)

    Hassan Motamed

    2013-01-01

    Full Text Available Hymenoptera stings are self-limiting events or due to allergic reactions. Sometimes envenomation with Hymenoptera can cause rare complications such as acute encephalopathy, peripheral neuritis, acute renal failure, nephrotic syndrome, silent myocardial infarction, rhabdomyolysis, conjunctivitis, corneal infiltration, lens subluxation, and optic neuropathy. The mechanism of peripheral nervous system damage is not clearly known. In our studied case after bee sting on face between the eyebrows with little erythema and  cm in size, bilateral blindness developed and gradually improved. Lateral movement of eyes was restricted with no pain. Involvement of cranial nerves including II, V, and VI was found. With conservative therapy after a year significant improvement has been achieved.

  8. Isolating gene-corrected stem cells without drug selection.

    Science.gov (United States)

    Hatada, Seigo; Arnold, Larry W; Hatada, Tomoko; Cowhig, John E; Ciavatta, Dominic; Smithies, Oliver

    2005-11-08

    Progress in isolating stem cells from tissues, or generating them from adult cells by nuclear transfer, encourages attempts to use stem cells from affected individuals for gene correction and autologous therapy. Current viral vectors are efficient at introducing transgenic sequences but result in random integrations. Gene targeting, in contrast, can directly correct an affected gene, or incorporate corrective sequences into a site free from undesirable side effects, but efficiency is low. Most current targeting procedures, consequently, use positive-negative selection with drugs, often requiring >/=10 days. This drug selection causes problems with stem cells that differentiate in this time or require feeder cells, because the feeders must be drug resistant and so are not eliminated by the selection. To overcome these problems, we have developed a procedure for isolating gene-corrected stem cells free from feeder cells after 3-5 days culture without drugs. The method is still positive-negative, but the positive and negative drug-resistance genes are replaced with differently colored fluorescence genes. Gene-corrected cells are isolated by FACS. We tested the method with mouse ES cells having a mutant hypoxanthine phosphoribosyltransferase (Hprt) gene and grown on feeder cells. After 5 days in culture, gene-corrected cells were obtained free from feeder cells at a "purity" of >30%, enriched >2,000-fold and with a recovery of approximately 20%. Corrected cells were also isolated singly for clonal expansion. Our FACS-based procedure should be applicable at small or large scale to stem cells that can be cultured (with feeder cells, if necessary) for >/=3 days.

  9. TB or not TB?: a case of isolated testicular TB with scrotal involvement.

    LENUS (Irish Health Repository)

    Bhargava, A

    2009-06-01

    Despite the genitourinary tract being the most common site affected by extrapulmonary TB, isolated testicular TB remains a rare clinical entity. In patients with co-morbidities such as hepatic impairment, treatment proves a challenge, as first-line hepatotoxic pharmaceuticals are contraindicated. Here, we report a case of isolated testicular TB with scrotal involvement, on a background of hepatic dysfunction.

  10. Social Isolation among Caregivers of Court-Involved Youths: A Qualitative Investigation

    Science.gov (United States)

    Forsbrey, April D.; Frabutt, James M.; Smith, Heather L.

    2005-01-01

    The authors used qualitative research methodology to examine the lives of caregivers of court-involved youths. Caregiver social isolation, including overall lack of support, lack of school support, and isolation from self, emerged as a salient theme across 7 domains. Implications for counselors are discussed, and brief descriptions of several…

  11. Isolated Cardiac Involvement in Primary Amyloidosis: Presenting as Sick Sinus Syndrome and Heart Failure

    OpenAIRE

    Pattanshetty, Deepak J.; Bhat,Pradeep K; Chamberlain, Wendy A.; Lyons, Matthew R.

    2013-01-01

    Cardiac amyloidosis is an infiltrative cardiomyopathy with a grave prognosis. Its clinical manifestations include restrictive cardiomyopathy, diastolic heart failure, conduction defects, and arrhythmias. Isolated cardiac involvement and significant conduction disturbances are reported very infrequently. We report a rare case of isolated cardiac involvement in primary amyloidosis, in a 76-year-old man who initially presented with sick sinus syndrome that necessitated permanent pacemaker insert...

  12. Bioprospection and selection of bacteria isolated from environments contaminated with petrochemical residues for application in bioremediation.

    Science.gov (United States)

    Cerqueira, Vanessa S; Hollenbach, Emanuel B; Maboni, Franciele; Camargo, Flávio A O; Peralba, Maria do Carmo R; Bento, Fátima M

    2012-03-01

    The use of microorganisms with hydrocarbon degrading capability and biosurfactant producers have emerged as an alternative for sustainable treatment of environmental passives. In this study 45 bacteria were isolated from samples contaminated with petrochemical residues, from which 21 were obtained from Landfarming soil contaminated with oily sludge, 11 were obtained from petrochemical industry effluents and 13 were originated directly from oily sludge. The metabolization capability of different carbon sources, growth capacity and tolerance, biosurfactant production and enzymes detection were determined. A preliminary selection carried out through the analysis of capability for degrading hydrocarbons showed that 22% of the isolates were able to degrade all carbon sources employed. On the other hand, in 36% of the isolates, the degradation of the oily sludge started within 18-48 h. Those isolates were considered as the most efficient ones. Twenty isolates, identified based on partial sequencing of the 16S rRNA gene, were pre-selected. These isolates showed ability for growing in a medium containing 1% of oily sludge as the sole carbon source, tolerance in a medium containing up to 30% of oily sludge, ability for biosurfactant production, and expression of enzymes involved in degradation of aliphatic and aromatic compounds. Five bacteria, identified as Stenotrophomonas acidaminiphila BB5, Bacillus megaterium BB6, Bacillus cibi, Pseudomonas aeruginosa, and Bacillus cereus BS20 were shown to be promising for use as inoculum in bioremediation processes (bioaugmentation) of areas contaminated with petrochemical residues since they can use oily sludge as the sole carbon source and produce biosurfactants.

  13. Selection on plant male function genes identifies candidates for reproductive isolation of yellow monkeyflowers.

    Directory of Open Access Journals (Sweden)

    Jan E Aagaard

    Full Text Available Understanding the genetic basis of reproductive isolation promises insight into speciation and the origins of biological diversity. While progress has been made in identifying genes underlying barriers to reproduction that function after fertilization (post-zygotic isolation, we know much less about earlier acting pre-zygotic barriers. Of particular interest are barriers involved in mating and fertilization that can evolve extremely rapidly under sexual selection, suggesting they may play a prominent role in the initial stages of reproductive isolation. A significant challenge to the field of speciation genetics is developing new approaches for identification of candidate genes underlying these barriers, particularly among non-traditional model systems. We employ powerful proteomic and genomic strategies to study the genetic basis of conspecific pollen precedence, an important component of pre-zygotic reproductive isolation among yellow monkeyflowers (Mimulus spp. resulting from male pollen competition. We use isotopic labeling in combination with shotgun proteomics to identify more than 2,000 male function (pollen tube proteins within maternal reproductive structures (styles of M. guttatus flowers where pollen competition occurs. We then sequence array-captured pollen tube exomes from a large outcrossing population of M. guttatus, and identify those genes with evidence of selective sweeps or balancing selection consistent with their role in pollen competition. We also test for evidence of positive selection on these genes more broadly across yellow monkeyflowers, because a signal of adaptive divergence is a common feature of genes causing reproductive isolation. Together the molecular evolution studies identify 159 pollen tube proteins that are candidate genes for conspecific pollen precedence. Our work demonstrates how powerful proteomic and genomic tools can be readily adapted to non-traditional model systems, allowing for genome-wide screens

  14. Isolated cardiac involvement in primary amyloidosis: presenting as sick sinus syndrome and heart failure.

    Science.gov (United States)

    Pattanshetty, Deepak J; Bhat, Pradeep K; Chamberlain, Wendy A; Lyons, Matthew R

    2013-01-01

    Cardiac amyloidosis is an infiltrative cardiomyopathy with a grave prognosis. Its clinical manifestations include restrictive cardiomyopathy, diastolic heart failure, conduction defects, and arrhythmias. Isolated cardiac involvement and significant conduction disturbances are reported very infrequently. We report a rare case of isolated cardiac involvement in primary amyloidosis, in a 76-year-old man who initially presented with sick sinus syndrome that necessitated permanent pacemaker insertion. Subsequent symptoms of heart failure led to additional evaluation, including an endomyocardial biopsy that revealed primary cardiac amyloidosis. Medical therapy improved the patient's symptoms, and he was discharged from the hospital in stable condition. In addition to discussing the patient's case, we review the relevant medical literature.

  15. Dexmedetomidine-Induced Contraction Involves CPI-17 Phosphorylation in Isolated Rat Aortas

    Science.gov (United States)

    Ok, Seong-Ho; Kwon, Seong-Chun; Baik, Jiseok; Hong, Jeong-Min; Oh, Jiah; Han, Jeong Yeol; Sohn, Ju-Tae

    2016-01-01

    Dexmedetomidine, a highly selective α-2 adrenoceptor agonist, produces vasoconstriction, which leads to transiently increased blood pressure. The goal of this study was to investigate specific protein kinases and the associated cellular signal pathways responsible for the increased calcium sensitization induced by dexmedetomidine in isolated rat aortas, with a particular focus on phosphorylation-dependent inhibitory protein of myosin phosphatase (CPI-17). The effect of Y-27632 and chelerythrine on the dexmedetomidine-induced intracellular calcium concentration ([Ca2+]i) and tension were assessed using fura-2-loaded aortic strips. The effects of rauwolscine, Y-27632, chelerythrine, and ML-7 hydrochloride on the dexmedetomidine-induced phosphorylation of CPI-17 or of the 20-kDa regulatory light chain of myosin (MLC20) were investigated in rat aortic vascular smooth muscle cells. The effects of rauwolscine, Y-27632, and chelerythrine on the membrane translocation of Rho-kinase and protein kinase C (PKC) phosphorylation induced by dexmedetomidine were assessed. Y-27632 and chelerythrine each reduced the slopes of the [Ca2+]i-tension curves of dexmedetomidine-induced contraction, and Y-27632 more strongly reduced these slopes than did chelerythrine. Rauwolscine, Y-27632, chelerythrine, and ML-7 hydrochloride attenuated the dexmedetomidine-induced phosphorylation of CPI-17 and MLC20. Taken together, these results suggest that dexmedetomidine-induced contraction involves calcium sensitization, which appears to be mediated by CPI-17 phosphorylation via Rho-kinase or PKC. PMID:27706026

  16. Environmental Concern and Involvement of Individuals in Selected Voluntary Associations

    Science.gov (United States)

    Schuett, Michael A.; Ostergren, David.

    2003-01-01

    The authors examined environmental concern and involvement of individuals in two voluntary associations, the International Mountain Bicycling Association (IMBA) and National Off-Highway Vehicle Conservation Council, Inc. (NOHVCC). Environmental concerns were assessed using the New Environmental Paradigm (NEP). When compared, results from 10 of 12…

  17. Isolation and identification of a yeast strain involved in the degradation of Marrakech olive mill wastewater

    Directory of Open Access Journals (Sweden)

    Nawal Fakharedine

    2011-12-01

    Full Text Available Yeasts are widely used in various sectors of biomedical research, biotechnologies and environmental contexts. During the treatment of Olive oil Mill Wastewater (OMW by yeasts, extensive degradation of the polyphenols responsible for the highly toxic nature of this effluent occurs. In this study, one yeast isolate was selected for its ability to use the phenolic compounds in OMW as the sole carbon source for growth. The treatment of OMW using this isolate reduced the levels of polyphenols by about 50% and the fats by over 80%. Analysis of the polyphenol pool by HPLC revealed the presence of 6 compounds, most of which had disappeared after 5 and 25 days of treatment. Using API ID 32C, the taxonomic study of the selected isolate was found to be 99.9% identical to Candida valida. However, the molecular data using the Rep-PCR technique showed 83.6% similarity to Pichia membranifaciens and 80.6% to Pichia anomala.

  18. Isolated cutaneous involvement in a child with nodal anaplastic large cell lymphoma

    Directory of Open Access Journals (Sweden)

    Vibhu Mendiratta

    2016-01-01

    Full Text Available Non-Hodgkin lymphoma is a common childhood T-cell and B-cell neoplasm that originates primarily from lymphoid tissue. Cutaneous involvement can be in the form of a primary extranodal lymphoma, or secondary to metastasis from a non-cutaneous location. The latter is uncommon, and isolated cutaneous involvement is rarely reported. We report a case of isolated secondary cutaneous involvement from nodal anaplastic large cell lymphoma (CD30 + and ALK + in a 7-year-old boy who was on chemotherapy. This case is reported for its unusual clinical presentation as an acute febrile, generalized papulonodular eruption that mimicked deep fungal infection, with the absence of other foci of systemic metastasis.

  19. Selective involvement of the amygdala in systemic lupus erythematosus.

    Directory of Open Access Journals (Sweden)

    Bart J Emmer

    2006-12-01

    Full Text Available BACKGROUND: Antibodies specifically affect the amygdala in a mouse model of systemic lupus erythematosus (SLE. The aim of our study was to investigate whether there is also specific involvement of the amygdala in human SLE. METHODS AND FINDINGS: We analyzed a group of 37 patients with neuropsychiatric SLE (NP-SLE, 21 patients with SLE, and a group of 12 healthy control participants with diffusion weighted imaging (DWI. In addition, in a subset of eight patients, plasma was available to determine their anti-NMDAR antibody status. From the structural magnetic resonance imaging data, the amygdala and the hippocampus were segmented, as well as the white and gray matter, and the apparent diffusion coefficient (ADC was retrieved. ADC values between controls, patients with SLE, and patients with NP-SLE were tested using analysis of variance with post-hoc Bonferroni correction. No differences were found in the gray or white matter segments. The average ADC in the amygdala of patients with NP-SLE and SLE (940 x 10(-6 mm2/s; p = 0.006 and 949 x 10(-6 mm2/s; p = 0.019, respectively was lower than in healthy control participants (1152 x 10(-6 mm2/s. Mann-Whitney analysis revealed that the average ADC in the amygdala of patients with anti-NMDAR antibodies (n = 4; 802 x 10(-6 mm2/s was lower (p = 0.029 than the average ADC of patients without anti-NMDAR antibodies (n = 4; 979 x 10(-6 mm2/s and also lower (p = 0.001 than in healthy control participants. CONCLUSIONS: This is the first study to our knowledge to observe damage in the amygdala in patients with SLE. Patients with SLE with anti-NMDAR antibodies had more severe damage in the amygdala compared to SLE patients without anti-NMDAR antibodies.

  20. Selective isolation of bacteria from soil with hydrophobic materials

    OpenAIRE

    Oku, Shota; Nishiyama, Masaya; Takao, Yuji

    2011-01-01

    Bacterial strains having a hydrophobic cell surface have often been considered as degraders of hydrophobic organic pollutants in soil. In this study, bacterial strains were isolated using hydrophobic materials from 12 soil samples, and their cell surface hydrophobicity was determined by evaluating their adherence to n-hexane. Bacterial strains isolated using polytetrafluoroethylene (PTFE) membrane were more hydrophobic on an average than those isolated with styrene-divinylbenzene (DVB) partic...

  1. Isolation and selection of local isolates of chitinolitic bacteria that potent to biocontrol of larva stadia of Aedes aegypti L.

    Directory of Open Access Journals (Sweden)

    SRI PUJIYANTO

    2008-01-01

    Full Text Available The dengue fever was the most dangerously epidemic in Indonesia, so it is important to prevent the population of Aedes aegypti L. The larva stages of Ae. aegypti had an exosceleton from chitin, so if its exosceleton had been degraded the larva would be died. Many bacteria had chitin degradation activities, and the bacteria had potential as a biocontrol for Ae. aegypti larva stages. This research was got many of the local chitinolitic bacteria that had been potential as a biocontrol for Ae. aegypti larva stages. The chitinolitic bacteria were isolated with selective agar medium. The sources of isolates were collected from Central Java and West Java water resources. Selected isolates was done to get the higher chitinolitic activity in mineral water medium. The isolate LMB1-5 was potential as a bioinsecticide for Ae. aegypti larva stages. This isolate decreased 86.7% of the larva population in 7 days.

  2. Seismic response analysis of coupled building involving MR damper and elastomeric base isolation

    Directory of Open Access Journals (Sweden)

    M.K. Shrimali

    2015-06-01

    The present study investigates the comparative performance of three proposed schemes of coupled building control involving Magnetorheological (MR damper and elastomeric base isolation, named as, Semiactive, Hybrid 1 and Hybrid 2. The results of numerical study showed that Hybrid controls are more effective in controlling the response as compared to Semiactive control. Further, influence of device parameters on control performance has been investigated through a parametric study.

  3. GABA selectively increases mucin-1 expression in isolated pig jejunum.

    Science.gov (United States)

    Braun, Hannah-Sophie; Sponder, Gerhard; Pieper, Robert; Aschenbach, Jörg R; Deiner, Carolin

    2015-11-01

    The inhibitory neurotransmitter GABA (γ-aminobutyric acid) is synthesized by glutamic acid decarboxylase, which is expressed in the central nervous system and in various other tissues including the intestine. Moreover, GABA can be ingested in vegetarian diets or produced by bacterial commensals in the gastrointestinal tract. As previous studies in lung have suggested a link between locally increased GABA availability and mucin 5AC production, the present study sought to test whether the presence or lack of GABA (and its precursor glutamine) has an effect on intestinal mucin expression. Porcine jejunum epithelial preparations were incubated with two different amounts of GABA or glutamine on the mucosal side for 4 h, and changes in the relative gene expression of seven different mucins, enzymes involved in mucin shedding, GABA B receptor, enzymes involved in glutamine/GABA metabolism, glutathione peroxidase 2, and interleukin 10 were examined by quantitative PCR (TaqMan(®) assays). Protein expression of mucin-1 (MUC1) was analyzed by Western blot. On the RNA level, only MUC1 was significantly up-regulated by both GABA concentrations compared with the control. Glutamine-treated groups showed the same trend. On the protein level, all treatment groups showed a significantly higher MUC1 expression than the control group. We conclude that GABA selectively increases the expression of MUC1, a cell surface mucin that prevents the adhesion of microorganisms, because of its size and negative charge, and therefore propose that the well-described positive effects of glutamine on enterocytes and intestinal integrity are partly attributable to effects of its metabolite GABA.

  4. Isolation and selection of Bradyrhizobium from the root nodules of ...

    African Journals Online (AJOL)

    Makmal Sains Rumpai

    2012-07-19

    Jul 19, 2012 ... For isolation of Bradyrhizobium, root nodules were collected from indigo .... Colonies were observed and recorded the frequency of different forms of colonies. .... present study were rod shaped, motile and Gram negative.

  5. Synthesis and purification of iodoaziridines involving quantitative selection of the optimal stationary phase for chromatography.

    Science.gov (United States)

    Boultwood, Tom; Affron, Dominic P; Bull, James A

    2014-05-16

    The highly diastereoselective preparation of cis-N-Ts-iodoaziridines through reaction of diiodomethyllithium with N-Ts aldimines is described. Diiodomethyllithium is prepared by the deprotonation of diiodomethane with LiHMDS, in a THF/diethyl ether mixture, at -78 °C in the dark. These conditions are essential for the stability of the LiCHI2 reagent generated. The subsequent dropwise addition of N-Ts aldimines to the preformed diiodomethyllithium solution affords an amino-diiodide intermediate, which is not isolated. Rapid warming of the reaction mixture to 0 °C promotes cyclization to afford iodoaziridines with exclusive cis-diastereoselectivity. The addition and cyclization stages of the reaction are mediated in one reaction flask by careful temperature control. Due to the sensitivity of the iodoaziridines to purification, assessment of suitable methods of purification is required. A protocol to assess the stability of sensitive compounds to stationary phases for column chromatography is described. This method is suitable to apply to new iodoaziridines, or other potentially sensitive novel compounds. Consequently this method may find application in range of synthetic projects. The procedure involves firstly the assessment of the reaction yield, prior to purification, by (1)H NMR spectroscopy with comparison to an internal standard. Portions of impure product mixture are then exposed to slurries of various stationary phases appropriate for chromatography, in a solvent system suitable as the eluent in flash chromatography. After stirring for 30 min to mimic chromatography, followed by filtering, the samples are analyzed by (1)H NMR spectroscopy. Calculated yields for each stationary phase are then compared to that initially obtained from the crude reaction mixture. The results obtained provide a quantitative assessment of the stability of the compound to the different stationary phases; hence the optimal can be selected. The choice of basic alumina, modified to

  6. Influence of selected Rhizoctonia solani isolates on sugar beet seedlings

    Directory of Open Access Journals (Sweden)

    Skonieczek Paweł

    2016-04-01

    Full Text Available From 2008 to 2010 the levels of sugar beet seedlings infection caused by Rhizoctonia solani were compared in laboratory tests. Seven sugar beet lines were tested: H56, H66, S2, S3, S4, S5 and S6 as well as three control cultivars: Carlos, Esperanza and Janosik. Sugar beet lines with tolerance to rhizoctoniosis and cultivars without tolerance were infected artificially by R. solani isolates: R1, R28a and R28b. These isolates belong to the second anastomosis group (AG, which is usually highly pathogenic to beet roots. The aim of the experiment was to test whether the tolerance of sugar beet genotypes to R. solani AG 2 prevents both root rot, and damping-off of seedlings, induced by the pathogen. Sugar beet lines tolerant to brown root rot in laboratory tests were significantly less sensitive to infection of the seedlings by R. solani AG 2 isolates in comparison to control cultivars. Rhizoctonia solani AG 2 isolates demonstrated considerable differences in pathogenicity against seedlings of sugar beet lines and cultivars. The strongest infection of sugar beet seedlings occurred with the isolate R28b. The greatest tolerance to infection by AG 2 isolates was found for the S5 and S3 breeding lines.

  7. Selection for autochthonous bifidobacteial isolates adapted to simulated gastrointestinal fluid

    Directory of Open Access Journals (Sweden)

    H Jamalifar

    2010-03-01

    Full Text Available "nBackground and the purpose of the study: Bifidobacterial strains are excessively sensitive to acidic conditions and this can affect their living ability in the stomach and fermented foods, and as a result, restrict their use as live probiotic cultures. The aim of the present study was to obtain bifidobacterial isolates with augmented tolerance to simulated gastrointestinal condition using cross-protection method. "nMethods: Individual bifidobacterial strains were treated in acidic environment and also in media containing bile salts and NaCl. Viability of the acid and acid-bile-NaCl tolerant isolates was further examined in simulated gastric and small intestine by subsequent incubation of the probiotic bacteria in the corresponding media for 120 min. Antipathogenic activities of the adapted isolates were compared with those of the original strains. "nResults and major conclusion: The acid and acid-bile-NaCl adapted isolates showed improved viabilities significantly (p<0.05 in simulated gastric fluid compared to their parent strains. The levels of reduction in bacterial count (Log cfu/ml of the acid and acid-bile-NaCl adapted isolates obtained in simulated gastric fluid ranged from 0.64-3.06 and 0.36-2.43 logarithmic units after 120 min of incubation. There was no significant difference between the viability of the acid-bile-NaCl-tolerant isolates and the original strains in simulated small intestinal condition except for Bifidobacterium adolescentis (p<0.05. The presence of 15 ml of supernatants of acid-bile-NaCl-adapted isolates and also those of the initial Bifidobacterium strains inhibited pathogenic bacterial growth for 24 hrs. Probiotic bacteria with improved ability to survive in harsh gastrointestinal environment could be obtained by subsequent treatment of the strains in acid, bile salts and NaCl environments.

  8. Infection Profiles of Selected Aquabirnavirus Isolates in CHSE Cells.

    Directory of Open Access Journals (Sweden)

    Amr A A Gamil

    Full Text Available The wide host range and antigenic diversity of aquabirnaviruses are reflected by the presence of a collection of isolates with different sero- and genotypic properties that have previously been classified as such. Differences in cytopathogenic mechanisms and host responses induced by these isolates have not been previously examined. In the present study, we investigated infection profiles induced by genetically and serologically closely related as well as distant isolates in-vitro. CHSE-214 cells were infected with either E1S (serotype A3, genogroup 3, VR-299 (serotype A1, genogroup 1, highly virulent Sp (TA or avirulent Sp (PT (serotype A2, genogroup 5. The experiments were performed at temperatures most optimum for each of the isolates namely 15°C for VR-299, TA and PT strains and 20°C for E1S. Differences in virus loads and ability to induce cytopathic effect, inhibition of protein synthesis, apoptosis, and induction of IFNa, Mx1, PKR or TNFα gene expression at different times post infection were examined. The results showed on one hand, E1S with the highest ability to replicate, induce apoptosis and IFNa gene expression while VR-299 inhibited protein synthesis and induced Mx1 and PKR gene expression the most. The two Sp isolates induced the highest TNFα gene expression but differed in their ability to replicate, inhibit protein synthesis, and induce gene expression, with TA being more superior. Collectively, these findings point towards the adaptation by different virus isolates to suit environments and hosts that they patronize. Furthermore, the results also suggest that genetic identity is not prerequisite to functional similarities thus results of one aquabirnavirus isolate cannot necessarily be extrapolated to another.

  9. Isolation of an aldehyde dehydrogenase involved in the oxidation of fluoroacetaldehyde to fluoroacetate in Streptomyces cattleya.

    Science.gov (United States)

    Murphy, C D; Moss, S J; O'Hagan, D

    2001-10-01

    Streptomyces cattleya is unusual in that it produces fluoroacetate and 4-fluorothreonine as secondary metabolites. We now report the isolation of an NAD(+)-dependent fluoroacetaldehyde dehydrogenase from S. cattleya that mediates the oxidation of fluoroacetaldehyde to fluoroacetate. This is the first enzyme to be identified that is directly involved in fluorometabolite biosynthesis. Production of the enzyme begins in late exponential growth and continues into the stationary phase. Measurement of kinetic parameters shows that the enzyme has a high affinity for fluoroacetaldehyde and glycoaldehyde, but not acetaldehyde.

  10. Isolation and characterizacion of enzymes involved in the biosynthesis of secondary metabolites with phytotherapeutic interest

    OpenAIRE

    Trapero Mozos, Almudena

    2014-01-01

    Isolation and characterization of enzymes involved in the biosynthesis of secondary metabolites with phytotherapeutic interest. Las plantas medicinales han sido utilizadas como primer recurso sanitario desde la antigüedad. En los últimos años el interés de la industria farmacéutica por los productos naturales o metabolitos secundarios provenientes de plantas medicinales, se ha incrementado considerablemente, entre las que se incluyen el azafrán. Se conoce como especia azafrán a los estigma...

  11. In Vitro Susceptibility of Mycobacterium ulcerans Isolates to Selected Antimicrobials

    Directory of Open Access Journals (Sweden)

    Enid Owusu

    2017-01-01

    Full Text Available Background. The current definitive treatment of Buruli ulcer with antibiotics makes the issue of antimicrobial drug resistance an unavoidable one. This is as a result of drug misuse by health personnel and patients’ noncompliance to treatment regimen. Monitoring of these factors and screening for new effective antimicrobials are crucial to effective management of Buruli ulcer disease. This study therefore investigated the inhibitory activity of some antibiotics against isolates of Mycobacterium ulcerans. Methods. Activity of eight antibiotics was tested against twelve M. ulcerans isolates (2 reference strains and 10 clinical isolates. The anti-M. ulcerans activities were determined by the agar dilution method and the minimum inhibitory concentrations (MICs were determined by the agar proportion method. Results. All antimicrobials investigated had activity against M. ulcerans isolates tested. The MICs ranged from 0.16 μg/mL to 2.5 μg/mL. Azithromycin recorded the highest inhibitory activity at a mean MIC of 0.39 μg/mL, whilst clofazimine a second-line antileprosy drug, recorded the lowest at a mean MIC of 2.19 μg/mL. Among the four antituberculosis drugs, rifampicin had the highest activity with a mean MIC of 0.81 μg/mL. Conclusion. Azithromycin could be considered as a lucrative alternative to existing treatment methods for inhibiting M. ulcerans in Ghana.

  12. Medial Prefrontal Cortex Is Selectively Involved in Response Selection Using Visual Context in the Background

    Science.gov (United States)

    Lee, Inah; Shin, Ji Yun

    2012-01-01

    The exact roles of the medial prefrontal cortex (mPFC) in conditional choice behavior are unknown and a visual contextual response selection task was used for examining the issue. Inactivation of the mPFC severely disrupted performance in the task. mPFC inactivations, however, did not disrupt the capability of perceptual discrimination for visual…

  13. Evaluation of three selective media for isolation of Aggregatibacter actinomycetemcomitans

    NARCIS (Netherlands)

    Rurenga, P.; Raangs, E.; Singadji, Z.; Wekema-Mulder, G.; Veloo, A. C. M.; van Winkelhoff, A. J.

    2013-01-01

    Background and Objective: Aggregatibacter actinomycetemcomitans is a pathogen in oral and nonoral infections. Detection and quantification of this pathogen can be performed using selective culture techniques. The aim of this study was to establish the efficacy of two known selective media in their a

  14. Evaluation of three selective media for isolation of Aggregatibacter actinomycetemcomitans

    NARCIS (Netherlands)

    Rurenga, P; Raangs, E; Singadji, Z; Wekema - Mulder, G; Veloo, A C M; van Winkelhoff, A J

    2013-01-01

    BACKGROUND AND OBJECTIVE: Aggregatibacter actinomycetemcomitans is a pathogen in oral and nonoral infections. Detection and quantification of this pathogen can be performed using selective culture techniques. The aim of this study was to establish the efficacy of two known selective media in their a

  15. New rules for visual selection: Isolating procedural attention.

    Science.gov (United States)

    Ramamurthy, Mahalakshmi; Blaser, Erik

    2017-02-01

    High performance in well-practiced, everyday tasks-driving, sports, gaming-suggests a kind of procedural attention that can allocate processing resources to behaviorally relevant information in an unsupervised manner. Here we show that training can lead to a new, automatic attentional selection rule that operates in the absence of bottom-up, salience-driven triggers and willful top-down selection. Taking advantage of the fact that attention modulates motion aftereffects, observers were presented with a bivectorial display with overlapping, iso-salient red and green dot fields moving to the right and left, respectively, while distracted by a demanding auditory two-back memory task. Before training, since the motion vectors canceled each other out, no net motion aftereffect (MAE) was found. However, after 3 days (0.5 hr/day) of training, during which observers practiced selectively attending to the red, rightward field, a significant net MAE was observed-even when top-down selection was again distracted. Further experiments showed that these results were not due to perceptual learning, and that the new rule targeted the motion, and not the color of the target dot field, and global, not local, motion signals; thus, the new rule was: "select the rightward field." This study builds on recent work on selection history-driven and reward-driven biases, but uses a novel paradigm where the allocation of visual processing resources are measured passively, offline, and when the observer's ability to execute top-down selection is defeated.

  16. Patterns of Parental Involvement in Selected OECD Countries: Cross-National Analyses of PISA

    Science.gov (United States)

    Hartas, Dimitra

    2015-01-01

    Using data from the Programme for International Student Assessment (PISA), patterns of parental involvement were examined in selected OECD countries. The findings showed that, irrespective of educational qualifications, parents were frequently involved in their children's learning at the start of primary school and at age 15. Cross-national…

  17. Effects of natural and sexual selection on adaptive population divergence and premating isolation in a damselfly.

    Science.gov (United States)

    Svensson, Erik I; Eroukhmanoff, Fabrice; Friberg, Magne

    2006-06-01

    The relative strength of different types of directional selection has seldom been compared directly in natural populations. A recent meta-analysis of phenotypic selection studies in natural populations suggested that directional sexual selection may be stronger in magnitude than directional natural selection, although this pattern may have partly been confounded by the different time scales over which selection was estimated. Knowledge about the strength of different types of selection is of general interest for understanding how selective forces affect adaptive population divergence and how they may influence speciation. We studied divergent selection on morphology in parapatric, natural damselfly (Calopteryx splendens) populations. Sexual selection was stronger than natural selection measured on the same traits, irrespective of the time scale over which sexual selection was measured. Visualization of the fitness surfaces indicated that population divergence in overall morphology is more strongly influenced by divergent sexual selection rather than natural selection. Courtship success of experimental immigrant males was lower than that of resident males, indicating incipient sexual isolation between these populations. We conclude that current and strong sexual selection promotes adaptive population divergence in this species and that premating sexual isolation may have arisen as a correlated response to divergent sexual selection. Our results highlight the importance of sexual selection, rather than natural selection in the adaptive radiation of odonates, and supports previous suggestions that divergent sexual selection promotes speciation in this group.

  18. Isolation of genes (nif/hup cosmids) involved in hydrogenase and nitrogenase activities in Rhizobium japonicum.

    Science.gov (United States)

    Hom, S S; Graham, L A; Maier, R J

    1985-03-01

    Recombinant cosmids containing a Rhizobium japonicum gene involved in both hydrogenase (Hup) and nitrogenase (Nif) activities were isolated. An R. japonicum gene bank utilizing broad-host-range cosmid pLAFR1 was conjugated into Hup- Nif- R. japonicum strain SR139. Transconjugants containing the nif/hup cosmid were identified by their resistance to tetracycline (Tcr) and ability to grow chemoautotrophically (Aut+) with hydrogen. All Tcr Aut+ transconjugants possessed high levels of H2 uptake activity, as determined amperometrically. Moreover, all Hup+ transconjugants tested possessed the ability to reduce acetylene (Nif+) in soybean nodules. Cosmid DNAs from 19 Hup+ transconjugants were transferred to Escherichia coli by transformation. When the cosmids were restricted with EcoRI, 15 of the 19 cosmids had a restriction pattern with 13.2-, 4.0-, 3.0-, and 2.5-kilobase DNA fragments. Six E. coli transformants containing the nif/hup cosmids were conjugated with strain SR139. All strain SR139 transconjugants were Hup+ Nif+. Moreover, one nif/hup cosmid was transferred to 15 other R. japonicum Hup- mutants. Hup+ transconjugants of six of the Hup- mutants appeared at a frequency of 1.0, whereas the transconjugants of the other nine mutants remained Hup-. These results indicate that the nif/hup gene cosmids contain a gene involved in both nitrogenase and hydrogenase activities and at least one and perhaps other hup genes which are exclusively involved in H2 uptake activity.

  19. Functional divergence of the NIP III subgroup proteins involved altered selective constraints and positive selection

    Directory of Open Access Journals (Sweden)

    Zhu Zhujun

    2010-11-01

    Full Text Available Abstract Background Nod26-like intrinsic proteins (NIPs that belong to the aquaporin superfamily are unique to plants. According to homology modeling and phylogenetic analysis, the NIP subfamily can be further divided into three subgroups with distinct biological functions (NIP I, NIP II, and NIP III. In some grasses, the NIP III subgroup proteins (NIP2s were demonstrated to be permeable to solutes with larger diameter, such as silicic acid and arsenous acids. However, to date there is no data-mining or direct experimental evidences for the permeability of such larger solutes for dicot NIP2s, although they exhibit similar three-dimensional structures as those in grasses. It is therefore intriguing to investigate the molecular mechanisms that drive the evolution of plant NIP2s. Results The NIP III subgroup is more ancient with a divergence time that predates the monocot-dicot split. The proliferation of NIP2 genes in modern grass species is primarily attributed to whole genome and segmental chromosomal duplication events. The structure of NIP2 genes is relatively conserved, possessing five exons and four introns. All NIP2s possess an ar/R filter consisting of G, S, G, and R, except for the cucumber CsNIP2;2, where a small G in the H2 is substituted with the bulkier C residue. Our maximum likelihood analysis revealed that NIP2s, especially the loop A (LA region, have undergone strong selective pressure for adaptive evolution. The analysis at the amino acid level provided strong statistical evidences for the functional divergence between monocot and dicot NIP III subgroup proteins. In addition, several SDPs (Specificity Determining Positions responsible for functional specificity were predicted. Conclusions The present study provides the first evidences of functional divergence between dicot and monocot NIP2s, and suggests that positive selection, as well as a radical shift of evolutionary rate at some critical amino acid sites is the primary

  20. Contributions of natural and sexual selection to the evolution of premating reproductive isolation: a research agenda.

    Science.gov (United States)

    Safran, Rebecca J; Scordato, Elizabeth S C; Symes, Laurel B; Rodríguez, Rafael L; Mendelson, Tamra C

    2013-11-01

    Speciation by divergent natural selection is well supported. However, the role of sexual selection in speciation is less well understood due to disagreement about whether sexual selection is a mechanism of evolution separate from natural selection, as well as confusion about various models and tests of sexual selection. Here, we outline how sexual selection and natural selection are different mechanisms of evolutionary change, and suggest that this distinction is critical when analyzing the role of sexual selection in speciation. Furthermore, we clarify models of sexual selection with respect to their interaction with ecology and natural selection. In doing so, we outline a research agenda for testing hypotheses about the relative significance of divergent sexual and natural selection in the evolution of reproductive isolation.

  1. Calcium carbonate mineralization: involvement of extracellular polymeric materials isolated from calcifying bacteria.

    Science.gov (United States)

    Ercole, Claudia; Bozzelli, Paola; Altieri, Fabio; Cacchio, Paola; Del Gallo, Maddalena

    2012-08-01

    This study highlights the role of specific outer bacterial structures, such as the glycocalix, in calcium carbonate crystallization in vitro. We describe the formation of calcite crystals by extracellular polymeric materials, such as exopolysaccharides (EPS) and capsular polysaccharides (CPS) isolated from Bacillus firmus and Nocardia calcarea. Organic matrices were isolated from calcifying bacteria grown on synthetic medium--in the presence or absence of calcium ions--and their effect on calcite precipitation was assessed. Scanning electron microscopy observations and energy dispersive X-ray spectrometry analysis showed that CPS and EPS fractions were involved in calcium carbonate precipitation, not only serving as nucleation sites but also through a direct role in crystal formation. The utilization of different synthetic media, with and without addition of calcium ions, influenced the biofilm production and protein profile of extracellular polymeric materials. Proteins of CPS fractions with a molecular mass between 25 and 70 kDa were overexpressed when calcium ions were present in the medium. This higher level of protein synthesis could be related to the active process of bioprecipitation.

  2. Pathogenesis of selective insulin resistance in isolated hepatocytes.

    Science.gov (United States)

    Cook, Joshua R; Langlet, Fanny; Kido, Yoshiaki; Accili, Domenico

    2015-05-29

    The development of insulin resistance (IR) in the liver is a key pathophysiologic event in the development of type 2 diabetes. Although insulin loses its ability to suppress glucose production, it largely retains its capacity to drive lipogenesis. This selective IR results in the characteristic hyperglycemia and dyslipidemia of type 2 diabetes. The delineation of two branched pathways of insulin receptor (InsR) signaling to glucose versus triglyceride production, one through FoxO and the other through SREBP-1c, provides a mechanism to account for this pathophysiological abnormality. We tested the complementary hypothesis that selective IR arises due to different intrinsic sensitivities of glucose production versus de novo lipogenesis to insulin as a result of cell-autonomous down-regulation of InsR number in response to chronic hyperinsulinemia. We demonstrate in mouse primary hepatocytes that chronic hyperinsulinemia abrogates insulin's inhibition of glucose production, but not its stimulation of de novo lipogenesis. Using a competitive inhibitor of InsR, we show that there is a 4-fold difference between levels of InsR inhibition required to cause resistance of glucose production versus lipogenesis to the actions of insulin. Our data support a parsimonious model in which differential InsR activation underlies the selective IR of glucose production relative to lipogenesis, but both processes require signaling through Akt1/2. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  3. Imaging of chronic recurrent multifocal osteomyelitis of childhood first presenting with isolated primary spinal involvement

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, S.E. [Department of Radiology, University Hospital of Bern, Inselspital, 3010, Bern (Switzerland); Heini, P.; Kalbermatten, D. [Department of Orthopedic Surgery, University Hospital of Bern, Inselspital, Bern (Switzerland); Sauvain, M.J. [Department of Pediatric Rheumatology, University Hospital of Bern, Inselspital, Bern (Switzerland); Stauffer, E. [Department of Pathology, University Hospital of Bern, Inselspital, Bern (Switzerland); Geiger, L. [Section Nuclear Medicine, Department of Radiology, University Hospital of Bern, Inselspital, Bern (Switzerland); Johnston, J.O. [Department of Orthopedic Surgery, University of California at San Francisco, San Francisco, California (United States); Roggo, A. [Department of Surgery, University Hospital of Bern, Inselspital, Bern (Switzerland); Steinbach, L.S. [Department of Radiology, University of California at San Francisco, San Francisco, California (United States)

    2003-06-01

    Initial presentation with primary spinal involvement in chronic recurrent multifocal osteomyelitis of childhood (CRMO) is rare. Our objective was to review the imaging appearances of three patients who had CRMO who initially presented with isolated primary spinal involvement.Design and patients The imaging, clinical, laboratory and histology findings of the three patients were retrospectively reviewed. Imaging included seven spinal MR imaging scans, one computed tomography scan, nine bone scans, two tomograms and 16 radiographs. These were reviewed by two musculoskeletal radiologists and a consensus view is reported. All three patients presented with atraumatic spinal pain and had extensive bone spinal pathology. The patients were aged 11, 13 and 12 years. There were two females and one male.Results and conclusions The initial patient had thoracic T6 and T8 vertebra plana. Bone scan showed additional vertebral body involvement. Follow-up was available over a 3 year period. The second patient had partial collapse of T9 and, 2 years later, of C6. Subsequently extensive multifocal disease ensued and follow-up was available over 8 years. The third patient initially had L3 inferior partial collapse and 1 year later T8 involvement with multifocal disease. Follow-up was available over 3 years. The imaging findings of the three patients include partial and complete vertebra plana with a subchondral line adjacent to endplates associated with bone marrow MR signal alterations. Awareness of the imaging appearances may help the radiologist to include this entity in the differential diagnosis in children who present with spinal pathology and no history of trauma. Histopathological examination excludes tumor and infection but with typical imaging findings may not always be necessary. (orig.)

  4. The Kalash genetic isolate: ancient divergence, drift, and selection.

    Science.gov (United States)

    Ayub, Qasim; Mezzavilla, Massimo; Pagani, Luca; Haber, Marc; Mohyuddin, Aisha; Khaliq, Shagufta; Mehdi, Syed Qasim; Tyler-Smith, Chris

    2015-05-01

    The Kalash represent an enigmatic isolated population of Indo-European speakers who have been living for centuries in the Hindu Kush mountain ranges of present-day Pakistan. Previous Y chromosome and mitochondrial DNA markers provided no support for their claimed Greek descent following Alexander III of Macedon's invasion of this region, and analysis of autosomal loci provided evidence of a strong genetic bottleneck. To understand their origins and demography further, we genotyped 23 unrelated Kalash samples on the Illumina HumanOmni2.5M-8 BeadChip and sequenced one male individual at high coverage on an Illumina HiSeq 2000. Comparison with published data from ancient hunter-gatherers and European farmers showed that the Kalash share genetic drift with the Paleolithic Siberian hunter-gatherers and might represent an extremely drifted ancient northern Eurasian population that also contributed to European and Near Eastern ancestry. Since the split from other South Asian populations, the Kalash have maintained a low long-term effective population size (2,319-2,603) and experienced no detectable gene flow from their geographic neighbors in Pakistan or from other extant Eurasian populations. The mean time of divergence between the Kalash and other populations currently residing in this region was estimated to be 11,800 (95% confidence interval = 10,600-12,600) years ago, and thus they represent present-day descendants of some of the earliest migrants into the Indian sub-continent from West Asia.

  5. Uremic encephalopathy with isolated brainstem involvement revealed by magnetic resonance image: a case report.

    Science.gov (United States)

    Jia, Li-Jing; Qu, Zhen-Zhen; Zhang, Xue-Qian; Tian, Yu-Juan; Wang, Ying

    2017-08-08

    Uremic Encephalopathy (UE) is a neurological complication associated with acute or chronic renal failure. Imaging findings of UE may present involvement of the basal ganglia, cortical or subcortical regions, and white matter. We report a rare case of UE caused by neurogenic bladder with isolated brainstem involvement revealed by magnetic resonance imaging (MRI). Immediate therapy resulted in full recovery of neurological signs and changes on MRI. A 14-year-old Han Chinese woman with a history of chronic renal failure caused by neurogenic bladder. On admission, she was unconscious and her pupils presented different sizes, while her vital signs were normal. MRI showed high signal in the dorsal pontine base and in the mid brain on fluid-attenuated inversion-recovery (FLAIR) imaging and on T2-weighted imaging while the signal was normal on diffusion-weighted images (DWI). Blood analysis revealed renal failure and acidosis. After urinary retention treatment and acidosis correction, the patient soon recovered. Follow-up MRI 2 months after the discharge revealed complete resolution of UE in the brainstem. We reported a rare case of a patient with UE that had unusual imaging manifestations for whom timely diagnosis and treatment assured recovery.

  6. Investigation of genes involved in nisin production in Enterococcus spp. strains isolated from raw goat milk.

    Science.gov (United States)

    Perin, Luana Martins; Todorov, Svetoslav Dimitrov; Nero, Luís Augusto

    2016-09-01

    Different strains of Lactococcus lactis are capable of producing the bacteriocin nisin. However, genetic transfer mechanisms allow the natural occurrence of genes involved in nisin production in members of other bacterial genera, such as Enterococcus spp. In a previous study, nisA was identified in eight enterococci capable of producing antimicrobial substances. The aim of this study was to verify the presence of genes involved in nisin production in Enterococcus spp. strains, as well as nisin expression. The nisA genes from eight Enterococcus spp. strains were sequenced and the translated amino acid sequences were compared to nisin amino-acid sequences previously described in databases. Although containing nisin structural and maturation related genes, the enterococci strains tested in the present study did not present the immunity related genes (nisFEG and nisI). The translated sequences of nisA showed some point mutations, identical to those presented by Lactococcus strains isolated from goat milk. All enterococci were inhibited by nisin, indicating the absence of immunity and thus that nisin cannot be expressed. This study demonstrated for the first time the natural occurrence of nisin structural genes in Enterococcus strains and highlights the importance of providing evidence of a link between the presence of bacteriocin genes and their expression.

  7. Selection of Beauveria bassiana and Metarhizium anisopliae Isolates to Control Triatoma infestans

    Directory of Open Access Journals (Sweden)

    Luz Christian

    1998-01-01

    Full Text Available Twenty three isolates of Beauveria bassiana and 13 isolates of Metarhizium anisopliae were tested on third instar nymphs of Triatoma infestans, a serious vector of Chagas disease. Pathogenicity tests at saturated humidity showed that this insect is very susceptible to fungal infection. At lower relative humidity (50%, conditions expected in the vector microhabitat, virulence was significantly different among isolates. Cumulative mortality 15 days after treatment varied from 17.5 to 97.5%, and estimates of 50% survival time varied from 6 to 11 days. Maintaining lower relative humidity, four B. bassiana and two M. anisopliae isolates were selected for analysis of virulence at different conidial concentrations and temperatures. Lethal concentrations sufficient to kill 50% of insects (LC50 varied from 7.1x105 to 4.3x106 conidia/ml, for a B. bassiana isolate (CG 14 and a M. anisopliae isolate (CG 491 respectively. Most isolates, particularly B. bassiana isolates CG 24 and CG 306, proved to be more virulent at 25 and 30°C, compared to 15 and 20°C. The differential virulence at 50% humidity observed among some B. bassiana isolates was not correlated to phenetic groups in cluster analysis of RAPD markers. In fact, the B. bassiana isolates analyzed presented a high homogeneity (> 73% similarity.

  8. Selective microbial genomic DNA isolation using restriction endonucleases.

    Science.gov (United States)

    Barnes, Helen E; Liu, Guohong; Weston, Christopher Q; King, Paula; Pham, Long K; Waltz, Shannon; Helzer, Kimberly T; Day, Laura; Sphar, Dan; Yamamoto, Robert T; Forsyth, R Allyn

    2014-01-01

    To improve the metagenomic analysis of complex microbiomes, we have repurposed restriction endonucleases as methyl specific DNA binding proteins. As an example, we use DpnI immobilized on magnetic beads. The ten minute extraction technique allows specific binding of genomes containing the DpnI Gm6ATC motif common in the genomic DNA of many bacteria including γ-proteobacteria. Using synthetic genome mixtures, we demonstrate 80% recovery of Escherichia coli genomic DNA even when only femtogram quantities are spiked into 10 µg of human DNA background. Binding is very specific with less than 0.5% of human DNA bound. Next Generation Sequencing of input and enriched synthetic mixtures results in over 100-fold enrichment of target genomes relative to human and plant DNA. We also show comparable enrichment when sequencing complex microbiomes such as those from creek water and human saliva. The technique can be broadened to other restriction enzymes allowing for the selective enrichment of trace and unculturable organisms from complex microbiomes and the stratification of organisms according to restriction enzyme enrichment.

  9. Parental Involvement, Selected Student Attributes, and Learning Outcomes in Instrumental Music.

    Science.gov (United States)

    Zdzinski, Stephen F.

    1996-01-01

    Discusses relationships among selected aspects of parental involvement as they relate to the cognitive, affective, and performance outcomes of instrumental music students. Discovers that for cognitive musical and musical performance outcomes parental influence is strongest at the elementary level. For affective outcomes parental involvement…

  10. A new selective enrichment procedure for isolating Pasteurella multocida from avian and environmental samples

    Science.gov (United States)

    Moore, M.K.; Cicnjak-Chubbs, L.; Gates, R.J.

    1994-01-01

    A selective enrichment procedure, using two new selective media, was developed to isolate Pasteurella multocida from wild birds and environmental samples. These media were developed by testing 15 selective agents with six isolates of P. multocida from wild avian origin and seven other bacteria representing genera frequently found in environmental and avian samples. The resulting media—Pasteurella multocida selective enrichment broth and Pasteurella multocida selective agar—consisted of a blood agar medium at pH 10 containing gentamicin, potassium tellurite, and amphotericin B. Media were tested to determine: 1) selectivity when attempting isolation from pond water and avian carcasses, 2) sensitivity for detection of low numbers of P. multocida from pure and mixed cultures, 3) host range specificity of the media, and 4) performance compared with standard blood agar. With the new selective enrichment procedure, P. multocida was isolated from inoculated (60 organisms/ml) pond water 84% of the time, whereas when standard blood agar was used, the recovery rate was 0%.

  11. An improved culture method for selective isolation of Campylobacter jejuni from wastewater

    Directory of Open Access Journals (Sweden)

    Jinyong Kim

    2016-08-01

    Full Text Available Campylobacter jejuni is one of the leading foodborne pathogens worldwide. C. jejuni is isolated from a wide range of foods, domestic animals, wildlife, and environmental sources. The currently-available culture-based isolation methods are not highly effective for wastewater samples due to the low number of C. jejuni in the midst of competing bacteria. To detect and isolate C. jejuni from wastewater samples, in this study, we evaluated a few different enrichment conditions using five different antibiotics (i.e., cefoperazone, vancomycin, trimethoprim, polymyxin B, and rifampicin, to which C. jejuni is intrinsically resistant. The selectivity of each enrichment condition was measured with Ct value using quantitative real-time PCR (qRT-PCR, and multiplex PCR to determine Campylobacter species. In addition, the efficacy of Campylobacter isolation on different culture media after selective enrichment was examined by growing on Bolton and Preston agar plates. The addition of polymyxin B, rifampicin, or both to the Bolton selective supplements enhanced the selective isolation of C. jejuni. In particular, rifampicin supplementation and an increased culture temperature (i.e., 42°C had a decisive effect on the selective enrichment of C. jejuni from wastewater. The results of 16S rDNA sequencing also revealed that Enterococcus spp. and Pseudomonas aeruginosa are major competing bacteria in the enrichment conditions. Although it is known to be difficult to isolate Campylobacter from samples with heavy contamination, this study well exhibited that the manipulation of antibiotic selective pressure improves the isolation efficiency of fastidious Campylobacter from wastewater.

  12. On selection and scaling of ground motions for analysis of seismically isolated structures

    Science.gov (United States)

    Pant, Deepak R.; Maharjan, Manika

    2016-12-01

    A broader consensus on the number of ground motions to be used and the method of scaling to be adopted for nonlinear response history analysis (RHA) of structures is yet to be reached. Therefore, in this study, the effects of selection and scaling of ground motions on the response of seismically isolated structures, which are routinely designed using nonlinear RHA, are investigated. For this purpose, isolation systems with a range of properties subjected to bidirectional excitation are considered. Benchmark response of the isolation systems is established using large sets of unscaled ground motions systematically categorized into pulse-like, non-pulse-like, and mixed set of motions. Different subsets of seven to 14 ground motions are selected from these large sets using (a) random selection and (b) selection based on the best match of the shape of the response spectrum of ground motions to the target spectrum. Consequences of weighted scaling (also commonly referred to as amplitude scaling or linear scaling) as well as spectral matching are investigated. The ground motion selection and scaling procedures are evaluated from the viewpoint of their accuracy, efficiency, and consistency in predicting the benchmark response. It is confirmed that seven time histories are sufficient for a reliable prediction of isolation system displacement demands, for all ground motion subsets, selection and scaling procedures, and isolation systems considered. If ground motions are selected based on their best match to the shape of the target response spectrum (which should be preferred over randomly selected motions), weighted scaling should be used if pulse-like motions are considered, either of weighted scaling or spectral matching can be used if non-pulse-like motions are considered, and an average of responses from weighted-scaled and spectrum-matched ground motions should be used for a mixed set of motions. On the other hand, the importance of randomly selected motions in

  13. Muscle magnetic resonance imaging in spinal muscular atrophy type 3: Selective and progressive involvement.

    Science.gov (United States)

    Durmus, Hacer; Yilmaz, Ravza; Gulsen-Parman, Yesim; Oflazer-Serdaroglu, Piraye; Cuttini, Marina; Dursun, Memduh; Deymeer, Feza

    2017-05-01

    In this study we sought to identify magnetic resonance imaging (MRI) signs of selective muscle involvement and disease progression in patients with spinal muscular atrophy type 3b (SMA3b). Twenty-five patients with genetically confirmed SMA3b underwent MRI on a 1.5-Tesla MR scanner. MRI showed significantly more severe involvement of the iliopsoas than of the gluteus maximus muscles, and more severe involvement of the triceps brachii than of the biceps brachii muscles. The quadriceps femoris muscles were severely involved. The deltoid, adductor longus, portions of the hamstrings, gracilis, sartorius, and rectus abdominis muscles were well preserved. We found a significant positive correlation between MRI changes and disease duration for gluteus maximus and triceps brachii. Follow-up MRIs of 4 patients showed disease progression. This study confirms the pattern of selective muscle involvement suggested by previous studies and further refines muscle MRI changes in SMA3b. Progressive muscle involvement is implicated. Muscle Nerve 55: 651-656, 2017. © 2016 Wiley Periodicals, Inc.

  14. Receptors involved in moxonidine-stimulated atrial natriuretic peptide release from isolated normotensive rat hearts.

    Science.gov (United States)

    Mukaddam-Daher, Suhayla; Menaouar, Ahmed; Gutkowska, Jolanta

    2006-07-10

    Imidazoline I1-receptors are present in the heart and may be involved in atrial natriuretic peptide (ANP) release. The following studies investigated whether moxonidine (an antihypertensive imidazoline I1-receptor and alpha2-adrenoceptor agonist) acts directly on the heart to stimulate ANP release, and to characterize the receptor type involved in this action. Perfusion of rat (200-225 g) isolated hearts with moxonidine (10(-6) and 10(-5) M), for 30 min, resulted in ANP release (83+/-29 and 277+/-70 ng/30 min, above basal, respectively), significantly (Palpha1-adrenoceptors), and prazosin (alpha1>alpha2-adrenoceptors), but increased by rauwolscine (alpha2-adrenoceptors). Perfusion with 10(-5) M brimonidine (full alpha2-adrenoceptor agonist) inhibited moxonidine-stimulated ANP release. Similarly, moxonidine (10(-6) M) tended to reduce coronary flow, but significantly increased coronary flow in the presence of brimonidine, which was vasoconstrictive when perfused alone. Coronary flow was reduced by 10(-5) M each, brimonidine>clonidine>moxonidine; while similar bradycardia was observed with clonidine and moxonidine, but not with brimonidine. In conclusion, these results argue in favor of moxonidine acting primarily on imidazoline I1-receptors to release ANP, with both alpha2-adrenoceptor and imidazoline I1-receptors exerting inhibitory inter-relation. In contrast, the coronary vasodilatory effect of moxonidine requires full activation of alpha2-adrenoceptor. The sympatholytic and ANP-releasing effects of moxonidine appear to be mediated by cardiac imidazoline receptors that may be differentially localized. Most importantly, moxonidine can stimulate ANP release from the heart without contribution of the central nervous system.

  15. Residual NADPH Oxidase Activity and Isolated Lung Involvement in X-Linked Chronic Granulomatous Disease

    Directory of Open Access Journals (Sweden)

    Maria J. Gutierrez

    2012-01-01

    Full Text Available Chronic granulomatous disease (CGD is characterized by inherited immune defects resulting from mutations in the NADPH oxidase complex genes. The X-linked type of CGD is caused by defects in the CYBB gene that encodes gp91-phox, a fundamental component of the NADPH oxidase complex. This mutation originates the most common and severe form of CGD, which typically has absence of NADPH oxidase function and aggressive multisystemic infections. We present the case of a 9-year-old child with a rare CYBB mutation that preserves some NADPH oxidase activity, resulting in an atypical mild form of X-linked CGD with isolated lung involvement. Although the clinical picture and partially preserved oxidase function suggested an autosomal recessive form of CGD, genetic testing demonstrated a mutation in the exon 3 of CYBB gene (c.252 G>A, p.Ala84Ala, an uncommon X-linked CGD variant that affects splicing. Atypical presentation and diagnostic difficulties are discussed. This case highlights that the diagnosis of mild forms of X-linked CGD caused by rare CYBB mutations and partially preserved NADPH function should be considered early in the evaluation of atypical and recurrent lung infections.

  16. Identification, isolation and expansion of myoendothelial cells involved in leech muscle regeneration.

    Directory of Open Access Journals (Sweden)

    Annalisa Grimaldi

    Full Text Available Adult skeletal muscle in vertebrates contains myoendothelial cells that express both myogenic and endothelial markers, and which are able to differentiate into myogenic cells to contribute to muscle regeneration. In spite of intensive research efforts, numerous questions remain regarding the role of cytokine signalling on myoendothelial cell differentiation and muscle regeneration. Here we used Hirudo medicinalis (Annelid, leech as an emerging new model to study myoendothelial cells and muscle regeneration. Although the leech has relative anatomical simplicity, it shows a striking similarity with vertebrate responses and is a reliable model for studying a variety of basic events, such as tissue repair. Double immunohistochemical analysis were used to characterize myoendothelial cells in leeches and, by injecting in vivo the matrigel biopolymer supplemented with the cytokine Vascular Endothelial Growth Factor (VEGF, we were able to isolate this specific cell population expressing myogenic and endothelial markers. We then evaluated the effect of VEGF on these cells in vitro. Our data indicate that, similar to that proposed for vertebrates, myoendothelial cells of the leech directly participate in myogenesis both in vivo and in vitro, and that VEGF secretion is involved in the recruitment and expansion of these muscle progenitor cells.

  17. Isolation of single-base genome-edited human iPS cells without antibiotic selection.

    Science.gov (United States)

    Miyaoka, Yuichiro; Chan, Amanda H; Judge, Luke M; Yoo, Jennie; Huang, Miller; Nguyen, Trieu D; Lizarraga, Paweena P; So, Po-Lin; Conklin, Bruce R

    2014-03-01

    Precise editing of human genomes in pluripotent stem cells by homology-driven repair of targeted nuclease-induced cleavage has been hindered by the difficulty of isolating rare clones. We developed an efficient method to capture rare mutational events, enabling isolation of mutant lines with single-base substitutions without antibiotic selection. This method facilitates efficient induction or reversion of mutations associated with human disease in isogenic human induced pluripotent stem cells.

  18. Debaryomyces hansenii strains from Valle de los Pedroches Iberian dry meat products: isolation, identification, characterization and selection for starter cultures.

    Science.gov (United States)

    Ramos, José; Melero, Yessica; Ramos-Moreno, Laura; Michán, Carmen; Cabezas, Lourdes

    2017-07-07

    Yeasts, filamentous fungi and bacteria colonize the surface of fermented sausages during the ripening process. The source of this microbiota is their surrounding environment, and it is influenced by their maturing conditions and by starter cultures. Debaryomyces hansenii has been previously isolated from several dry-cured meat products, and it has been associated with the lipolytic and proteolytic changes that occur in these products, which influence their taste and flavour. In this work, the yeast microbiota present in casing from different meat products ("lomo", "chorizo" and "salchichón") from the Valle de los Pedroches region, South-Spain, were isolated for the first time. D. hansenii was by far the most abundant species in these products, as all 22 different isolates selected were identified as D. hansenii by biochemical and/or molecular methods. In contrast, no yeasts were found in the meat batter. Our data constitute the first study on the yeasts present in "lomo" sausages and particularly on the highly appreciated "Valle de los Pedroches" ones. Furthermore, the resistance of these isolates to different pH, temperatures and saline stress was studied, together with their catabolic characteristics. Based on those results, some isolates are proposed as valuable candidates for starter cultures that could improve both the manufacture and the flavor of this dry-cured meat products, together with the understanding of new mechanisms involved in stress tolerance. Applied medium-scale industrial tests are currently in progress.

  19. Membrane permeability, a pivotal function involved in antibiotic resistance and virulence in Enterobacter aerogenes clinical isolates.

    Science.gov (United States)

    Lavigne, J-P; Sotto, A; Nicolas-Chanoine, M-H; Bouziges, N; Bourg, G; Davin-Regli, A; Pagès, J-M

    2012-06-01

    Imipenem-susceptible E. aerogenes isolates exhibiting extended spectrum β-lactamases, target mutations and a basal efflux expression, were identified in five patients. After imipenem treatment, imipenem-intermediate susceptible (IMI-I) or resistant (IMI-R) isolates emerged in these patients. Alteration in porin synthesis and increase in efflux expression were observed in the IMI-I isolates whereas complete loss of the porins, LPS alteration and efflux overexpression were observed in the IMI-R isolates. Bacterial virulence of the strains was investigated by the Caenorhabditis elegans model. The IMI-R isolates were shown to be significantly less virulent than the IMI-susceptible or IMI-I isolates. The pleiotropic membrane alteration and its associated fitness burden exhibited by E. aerogenes isolates influence their antibiotic resistance and their virulence behaviour. These findings highlight the balance between the low permeability-related resistance and virulence and their relationships with the treatment of resistant pathogens.

  20. Selection of oleuropein-degrading lactic acid bacteria strains isolated from fermenting Moroccan green olives

    Energy Technology Data Exchange (ETDEWEB)

    Ghabbour, N.; Lamzira, Z.; Thonart, P.; Cidalia, P.; Markaouid, M.; Asehraoua, A.

    2011-07-01

    A total of 177 strains of lactic acid bacteria (LAB) were isolated from early-stage Moroccan Picholine green olive fermentation, including Lactobacillus plantarum (44.63%), Lactobacillus pentosus (25.99%), Lactobacillus brevis (9.61%) and Pediococcus pentosaceus (19.77%). All the isolates were screened for their tolerance to olive leaf extract and oleuropein. Most of the isolates (85.3%) were found able to degrade oleuropein, when evaluated by either oleuropein or 5-Bromo-4-chloro-3-indolyl {beta}-D-glucuronide (X-Gluc) as substrates. The biodegradation capacity of the selected strains of each species was confirmed by HPLC analysis. (Author).

  1. Isolation and characterization of Acinetobacter baumannii recovered from Campylobacter selective medium.

    Directory of Open Access Journals (Sweden)

    Dinesh M Fernando

    2016-11-01

    Full Text Available Acinetobacter baumannii, a Gram-negative opportunistic pathogen, is known to cause multidrug resistant infections. This organism has primarily been isolated from clinical environments and its environmental reservoirs remain largely unknown. In the present study, we recovered seven isolates of A. baumannii growing under conditions selective for Campylobacter spp. (microaerophilic at 42 oC and in the presence of antibiotics from dairy cattle manure storage tank or surface water impacted by livestock effluents. Antibiotic susceptibility tests revealed that all of these isolates were less susceptible to at least two different clinically relevant antibiotics, compared to the type strain A. baumannii ATCC17978. Expression of resistance-nodulation-division efflux pumps, an important mechanism of intrinsic resistance in these organisms, was analyzed and adeB was found to be overexpressed in one and adeJ was overexpressed in three isolates. Comparison of these isolates using genomic DNA Macro-Restriction Fragment Pattern Analysis (MRFPA revealed relatively low relatedness among themselves or with some of the clinical isolates from previous studies. This study suggests that A. baumannii isolates are capable of growing under selective conditions for Campylobacter spp. and that this organism can be present in manure and water.

  2. Selection Model of Optional Parts in Suppliers Involved in New Product Development

    Institute of Scientific and Technical Information of China (English)

    MU Lifeng; TANG Jiafu; PAN Zhendong; HAN Yi

    2006-01-01

    Quality function deployment (QFD), which can translate efficiently customer requirement (CRs) into technical attributes (TAs), is widely used in new product development (NPD) as an efficient customer-driven approach. After lots of QFD application is applied in manufacturing fields, it was proved that without considering suppliers involved, it is difficult to get precise and satisfying results in each process of QFD, particularly part deployment. The paper will address the issues of a model of optional parts selection in suppliers involved in part deployment process. Further, it has been applied to an ERP system reconstruction and updating problem as an illustration.

  3. Genetic diversity and positive selection analysis of classical swine fever virus isolates in south China.

    Science.gov (United States)

    Shen, Haiyan; Pei, Jingjing; Bai, Jialin; Zhao, Mingqiu; Ju, Chunmei; Yi, Lin; Kang, Yanmei; Zhang, Xuetao; Chen, Lijun; Li, Yinguang; Wang, Jiaying; Chen, Jinding

    2011-10-01

    Classical swine fever virus (CSFV) causes a highly contagious disease that leads to significant economic losses in the pig industry worldwide. However, there is a paucity of knowledge on the accurate genotyping of CSFV isolates in south China. This study genotyped the E2 gene of 14 CSFV strains isolated during 2008-2010 from domestic pigs in different districts of south China. Phylogenetic analyses revealed that all of the 14 CSFV isolates were clustered into genetic subgroup 1.1. This contrasts with most parts of China, where group 2 isolates are predominant. Furthermore, the positive selection pressures acting on the E(rns) and E2 envelope protein genes of CSFV were assessed and a site-by-site analysis of the dN/dS ratio was performed to identify specific codons that undergo diversification under positive selection. While no significant evidence for positive selection was observed in E(rns), two positively selected sites at amino acid residues 49 and 72 in the E2 encoding region were identified. Our results revealed that a predominance of subgroup 1.1 CSFV isolates is currently circulating in some districts of south China, which appear to be unrelated to the Chinese C-strain vaccine. Moreover, the envelope protein gene, E2, has undergone positive selection in 14 CSFV strains and two positively selected sites have been identified in this study. Understanding the molecular epidemiology and functional importance of these positively selected amino acid positions could help to predict possible changes in virulence, the development of vaccines and disease control.

  4. Selection of indigenous isolates of entomopathogenic soil fungus Metarhizium anisopliae under laboratory conditions.

    Science.gov (United States)

    Skalický, Aleš; Bohatá, Andrea; Šimková, Jana; Osborne, Lance S; Landa, Zdeněk

    2014-07-01

    Eight native isolates of the entomopathogenic fungus Metarhizium anisopliae (Metschnikoff) Sorokin were obtained by monitoring soils cultivated in a conventional manner. These isolates were compared in three areas: (a) conidial germination, (b) radial growth and sporulation and (c) ability of conidia to infect Tenebrio molitor larvae. All bioassays were carried out at constant temperatures of 10, 15, and 20 °C. Conidia of individual isolates demonstrated differences in germination after a 24-h long incubation at all evaluated temperatures. At 20 °C, the germination ranged from 67 to 100 % and at 15 °C from 5.33 to 46.67 %. At 10 °C, no germination was observed after 24 h; nevertheless, it was 8.67-44.67 % after 48 h. In terms of radial growth, the culture diameters and the associated production of spores of all isolates increased with increasing temperature. At 10 °C, sporulation was observed in three isolates while all remaining cultures appeared sterile. Three weeks post-inoculation, conidia of all assessed isolates caused 100 % cumulative mortality of treated larvae of T. molitor at 15 and 20 °C with the exception of isolate 110108 that induced 81.33 % mortality at 15 °C. At 10 °C, larval cumulative mortality ranged from 6.67 to 85.33 % depending on the isolate. Isolates 110108 and 110111 showed significantly slower outset and a much lower rate of infection at all temperatures compared to other tested isolates of M. anisopliae. The bioassays were carried out with the purpose to sort and select indigenous isolates of M. anisopliae useful as biocontrol agents in their original habitat.

  5. Isolation of cells for selective treatment and analysis using a magnetic microfluidic chip

    KAUST Repository

    Yassine, O.

    2014-05-01

    This study describes the development and testing of a magnetic microfluidic chip (MMC) for trapping and isolating cells tagged with superparamagnetic beads (SPBs) in a microfluidic environment for selective treatment and analysis. The trapping and isolation are done in two separate steps; first, the trapping of the tagged cells in a main channel is achieved by soft ferromagnetic disks and second, the transportation of the cells into side chambers for isolation is executed by tapered conductive paths made of Gold (Au). Numerical simulations were performed to analyze the magnetic flux and force distributions of the disks and conducting paths, for trapping and transporting SPBs. The MMC was fabricated using standard microfabrication processes. Experiments were performed with E. coli (K12 strand) tagged with 2.8 μm SPBs. The results showed that E. coli can be separated from a sample solution by trapping them at the disk sites, and then isolated into chambers by transporting them along the tapered conducting paths. Once the E. coli was trapped inside the side chambers, two selective treatments were performed. In one chamber, a solution with minimal nutrition content was added and, in another chamber, a solution with essential nutrition was added. The results showed that the growth of bacteria cultured in the second chamber containing nutrient was significantly higher, demonstrating that the E. coli was not affected by the magnetically driven transportation and the feasibility of performing different treatments on selectively isolated cells on a single microfluidic platform.

  6. Selective extraction of isolated mitotic apparatus. Evidence that typical microtubule protein is extracted by organic mercurial.

    Science.gov (United States)

    Bibring, T; Baxandall, J

    1971-02-01

    Mitotic apparatus isolated from sea urchin eggs has been treated with meralluride sodium under conditions otherwise resembling those of its isolation. The treatment causes a selective morphological disappearance of microtubules while extracting a major protein fraction, probably consisting of two closely related proteins, which constitutes about 10% of mitotic apparatus protein. Extraction of other cell particulates under similar conditions yields much less of this protein. The extracted protein closely resembles outer doublet microtubule protein from sea urchin sperm tail in properties considered typical of microtubule proteins: precipitation by calcium ion and vinblastine, electrophoretic mobility in both acid and basic polyacrylamide gels, sedimentation coefficient, molecular weight, and, according to a preliminary determination, amino acid composition. An antiserum against a preparation of sperm tail outer doublet microtubules cross-reacts with the extract from mitotic apparatus. On the basis of these findings it appears that microtubule protein is selectively extracted from isolated mitotic apparatus by treatment with meralluride, and is a typical microtubule protein.

  7. ISOLATED INVOLVEMENT OF CERVICAL LYMPH NODES IN CASTLEMAN’S DISEASE IN A YOUNG PATIENT: A RARE PRESENTATION

    Directory of Open Access Journals (Sweden)

    Madhu

    2016-05-01

    Full Text Available Isolated cervical nodal involvement of Castleman’s disease is very rare, accounting for 6% of all cases. Of the two histopathological variants of Castleman’s disease, the hyaline-vascular type is more common as seen in our patient and carries a better prognosis than the plasma cell type.

  8. Isolated Cu2+ ions: active sites for selective catalytic reduction of NO

    NARCIS (Netherlands)

    Korhonen, S.T.; Fickel, D.W.; Lobo, R.F.; Weckhuysen, B.M.; Beale, A.M.

    2011-01-01

    Cu chabazite catalysts show remarkable low temperature activity in selective catalytic reduction (SCR) of NO. This high activity is due to the unique character of the zeolite framework that allows only the presence of one type of isolated mononuclear Cu2+ species. These Cu2+ species are the active s

  9. Fractionation of five technical lignins by selective extraction in green solvents and characterization of isolated fractions

    NARCIS (Netherlands)

    Boeriu, C.G.; Fitigau, F.; Gosselink, R.J.A.; Frissen, A.E.; Stoutjesdijk, J.H.; Peter, F.

    2014-01-01

    Lignins from softwood, hardwood, grass and wheat straw were fractionated by selective extraction at ambient temperature using green solvents like acetone/water solutions of 10, 30, 50, 70 and 90% (v/v) acetone and ethyl acetate. A comparison between the isolated fractions and unfractionated lignins

  10. [A selective nutrient medium for isolating clinical strains of Escherichia coli O157:H7].

    Science.gov (United States)

    Sultanov, Z Z; Stepanova, E D; Kakulina, E A

    2000-01-01

    A dried selective culture medium, electrolyte-deficient sorbitol agar (EDS agar), for the isolation and preliminary identification of E. coli O157:H7 from clinical material has been developed. The medium is not inferior in its quality to analogous foreign media and requires no scarce ingredients for its manufacture.

  11. Identification of bap and icaA genes involved in biofilm formation in coagulase negative staphylococci isolated from feline conjunctiva.

    Science.gov (United States)

    Płoneczka-Janeczko, Katarzyna; Lis, Paweł; Bierowiec, Karolina; Rypuła, Krzysztof; Chorbiński, Paweł

    2014-12-01

    Bap and icaA genes are commonly known to be involved in the biofilm formation. The prevalence of bap and icaA genes and biofilm formation was determined in conjunctival isolates of coagulase negative staphylococci (CNS) collected from cats. The study was conducted on 90 archival CNS isolates collected from feline conjunctiva obtained from clinically healthy cats and cats with ocular problems. Biofilm formation was examined using the microtiter plate (MTP) method. The prevalence of icaA and bap genes was determined using polymerase chain reaction (PCR). Genetic profiles of the bap-positive isolates were examined using the modified random amplified polymorphic DNA (RAPD) method. Of the 90 CNS isolates investigated, 58.9% (53/90) were confirmed to form biofilms on a polystyrene plate after 24 h, and the intensity of the biofilm production varied strongly between positive strains. Among the biofilm-producing isolates, 24.5% (13/53) carried the icaA gene and 3.8% (2/53) carried the bap gene. Among the isolates that did not produce biofilms, the icaA gene and bap gene were detected in 8.1% (3/37) and 2.7% (1/37) of isolates, respectively. This is the first report demonstrating that CNS isolated from feline conjunctiva can potentially be a bap gene reservoir. Preliminary comparison of the genetic profiles of three bap-positive isolates collected from cats showed that each of the isolates has a different genetic background with a high similarity with the human strain of S. epidermidis.

  12. Esophageal Cicatricial Pemphigoid as an Isolated Involvement Treated with Mycophenolate Mofetil

    Directory of Open Access Journals (Sweden)

    Sandra Sánchez Prudencio

    2015-01-01

    Full Text Available Cicatricial pemphigoid (CP is a rare blistering autoimmune disease. Esophageal involvement occurs in widespread disease and rarely appears as the only affected organ. We report a 67-year-old Caucasian female with esophageal dysphagia and weight loss. Several oral panendoscopies showed multiple exudative ulcerations with fibrin and webs in mid- and proximal esophagus and a peeling mucosa. There were no lesions in other organs. We established the diagnosis performing a direct immunofluorescence (DIF, demonstrating IgG3 and complement deposition along the basement membrane. As initial treatment the patient received prednisone 60 mg and 1 gr twice daily of mycophenolate mofetil (MMF as a steroid-sparing agent due to its lower toxicity and its selective mechanism of action. Six months later there was a significant clinical improvement and the esophageal ulcerations had disappeared, developing cicatricial fibrous rings, although no stenosis was present. Four years later, the patient remains asymptomatic with a low maintenance dose of MMF.

  13. Actuarial risk of isolated CNS involvement in Ewing's sarcoma following prophylactic cranial irradiation and intrathecal methotrexate

    Energy Technology Data Exchange (ETDEWEB)

    Trigg, M.E.; Makuch, R.; Glaubiger, D.

    1985-04-01

    Records of 154 patients with Ewing's sarcoma treated at the National Cancer Institute were reviewed to assess the incidence and risk of developing isolated central nervous system (CNS) Ewing's sarcoma. Sixty-two of the 154 patients had received CNS irradiation and intrathecal (i.t.) methotrexate as part of their initial therapy to prevent the occurrence of isolated CNS Ewing's sarcoma. The risk of developing isolate CNS Ewing's sarcoma was greatest within the first two years after diagnosis and was approximately 10%. The overall risk of CNS recurrence in the group of patients receiving DNS treatment was similar to the group receiving no therapy directed to the CNS. The occurrence of isolated CNS involvement was not prevented by the use of CNS irradiation and i.t. methotrexate. Because of a lack of efficacy to the CNS irradiation regimen, current treatment regimens do not include therapy directed to CNS.

  14. Fungicide sensitivity of selected Verticillium fungicola isolates from Agaricus bisporus farms

    Directory of Open Access Journals (Sweden)

    Potočnik Ivana

    2008-01-01

    Full Text Available Five isolates of Verticillium fungicola, isolated from diseased fruiting bodies of Agaricus bisporus collected from mushroom farms in Serbia during 2002-2003, were studied. By observing their colony morphology under different growth conditions and their pathogenic characteristics, the isolates were identified as V. fungicola var. fungicola. The peat/lime casing was the primary source of infection. Testing of sensitivity to selected fungicides showed that all isolates were highly resistant to benomyl (EC50 values were higher than 200.00 mg/l, moderately sensitive to iprodione (EC50 values were between 11.93 and 22.80 mg/l, and highly sensitive to prochloraz-Mn (EC50 values were less than 3.00 mg/l.

  15. Isolates of citrus exocortis viroid recovered by host and tissue selection.

    Science.gov (United States)

    Semancik, J S; Szychowski, J A; Rakowski, A G; Symons, R H

    1993-11-01

    Isolates of citrus exocortis viroid (CEV) from a single sweet orange citrus source have been selected by sequential passage through the alternative hosts citron, Gynura aurantiaca, a hybrid tomato Lycopersicon esculentum x L. peruvianum, and from disorganized callus culture of the hybrid tomato. The distinctions in symptom expression, titre and electrophoretic mobility among the CEV isolates, operationally termed CEVc (citron), CEVg (Gynura), CEVt (tomato) and CEVcls (callus) are supported by characteristically different nucleotide sequences. The nucleotide sequence of full-length cDNA clones of CEVc purified from citron shows exchanges not reported for any previously described CEV variant. An unusual number of exchanges have been localized in the terminal domains of all the isolates analysed here. A common pattern of nucleotide exchanges, described as a 'tomato signature', can be detected in all of the isolates derived from hybrid tomato tissues.

  16. Clonality of Providencia stuartii isolates involved in outbreak that occurred in a burn unit.

    Science.gov (United States)

    Saida, N Ben; Thabet, L; Messadi, A; Bouselmi, K; Turki, A; Boukadida, J

    2008-09-01

    In order to investigate an outbreak of multidrug-resistant Providencia stuartii that occurred in a burn unit, we analyzed by pulsed-field gel electrophoresis (PFGE) all isolates of P. stuartii collected during 4 months of 2005 from patients and from a tracheal aspirator. Seventeen clinical isolates of P. stuartii, extended-spectrum beta-lactamase (ESBL) producing, were collected from 17 patients. All these isolates were nosocomially acquired. Three other isolates were collected from the aspirator probe, the aspirator reservoir and from the aspirator tube. Three different antibiotypes were identified without correlation with the genotype. Two PFGE types were obtained (types A and B) with predominance of one (type A) that was observed for 15 isolates. P. stuartii isolates collected from different components of the aspirator (probe, reservoir and tube) yielded PFGE type A. This study suggests the bi-clonality of the outbreak and that transmission of epidemic P. stuartii isolates was through a common source. The aspirator probe, contaminated from aspirator that functioned as a reservoir of bacteria, seems to be the route of transmission of P. stuartii. Furthermore, this study shows the utility of PFGE in typing for the purpose of understanding the epidemiological behaviour of P. stuartii and as a basis for the development of rational control strategies.

  17. Involvement of prefrontal AMPA receptors in encounter stimulation-induced hyperactivity in isolation-reared mice.

    Science.gov (United States)

    Araki, Ryota; Ago, Yukio; Hasebe, Shigeru; Nishiyama, Saki; Tanaka, Tatsunori; Oka, Satoshi; Takuma, Kazuhiro; Matsuda, Toshio

    2014-06-01

    We recently showed that social encounter stimulation induces hyperactivity in mice reared in social isolation from early life and this is associated with the transient activation of prefrontal dopaminergic and serotonergic systems. In the present study, we examined the effect of the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptor antagonist 2, 3-dioxo-6-nitro-1, 2, 3, 4-tetrahydrobenzo[f]quinoxaline-7-sulfonamide (NBQX) on encounter-induced behavioural and neurochemical changes to study the role of the receptor in abnormal behaviours in isolation-reared mice. The encounter to an intruder mouse induced hyperactivity with transient increases in prefrontal dopamine and serotonin levels in isolation-reared mice. NBQX attenuated the encounter-induced hyperactivity and the associated neurochemical changes in isolation-reared mice. In addition, NBQX reduced aggressive behaviour and cognitive impairment in isolation-reared mice, but did not affect depressive-like behaviour or spontaneous hyper-locomotion in these animals. The AMPA receptor agonist (S)-AMPA increased prefrontal dopamine and serotonin release, and this effect was higher in isolation-reared mice than in the group-reared mice, suggesting higher prefrontal AMPA receptor activity in isolation-reared mice. Furthermore, isolation rearing increased the expression of AMPA receptor subunits (GluR1, GluR2 and GluR3) and GluR1 Ser845 phosphorylation in the prefrontal cortex, but not in the hippocampus or nucleus accumbens. Taken together, these results suggest that an increase in AMPA receptor activity in the prefrontal cortex contributes to some, but not all, abnormal behaviours in isolation-reared mice.

  18. Genetic polymorphism and natural selection of Duffy binding protein of Plasmodium vivax Myanmar isolates.

    Science.gov (United States)

    Ju, Hye-Lim; Kang, Jung-Mi; Moon, Sung-Ung; Kim, Jung-Yeon; Lee, Hyeong-Woo; Lin, Khin; Sohn, Woon-Mok; Lee, Jin-Soo; Kim, Tong-Soo; Na, Byoung-Kuk

    2012-03-01

    Plasmodium vivax Duffy binding protein (PvDBP) plays an essential role in erythrocyte invasion and a potential asexual blood stage vaccine candidate antigen against P. vivax. The polymorphic nature of PvDBP, particularly amino terminal cysteine-rich region (PvDBPII), represents a major impediment to the successful design of a protective vaccine against vivax malaria. In this study, the genetic polymorphism and natural selection at PvDBPII among Myanmar P. vivax isolates were analysed. Fifty-four P. vivax infected blood samples collected from patients in Myanmar were used. The region flanking PvDBPII was amplified by PCR, cloned into Escherichia coli, and sequenced. The polymorphic characters and natural selection of the region were analysed using the DnaSP and MEGA4 programs. Thirty-two point mutations (28 non-synonymous and four synonymous mutations) were identified in PvDBPII among the Myanmar P. vivax isolates. Sequence analyses revealed that 12 different PvDBPII haplotypes were identified in Myanmar P. vivax isolates and that the region has evolved under positive natural selection. High selective pressure preferentially acted on regions identified as B- and T-cell epitopes of PvDBPII. Recombination may also be played a role in the resulting genetic diversity of PvDBPII. PvDBPII of Myanmar P. vivax isolates displays a high level of genetic polymorphism and is under selective pressure. Myanmar P. vivax isolates share distinct types of PvDBPII alleles that are different from those of other geographical areas. These results will be useful for understanding the nature of the P. vivax population in Myanmar and for development of PvDBPII-based vaccine.

  19. Population genomics of the killer whale indicates ecotype evolution in sympatry involving both selection and drift.

    Science.gov (United States)

    Moura, Andre E; Kenny, John G; Chaudhuri, Roy; Hughes, Margaret A; J Welch, Andreanna; Reisinger, Ryan R; de Bruyn, P J Nico; Dahlheim, Marilyn E; Hall, Neil; Hoelzel, A Rus

    2014-11-01

    The evolution of diversity in the marine ecosystem is poorly understood, given the relatively high potential for connectivity, especially for highly mobile species such as whales and dolphins. The killer whale (Orcinus orca) has a worldwide distribution, and individual social groups travel over a wide geographic range. Even so, regional populations have been shown to be genetically differentiated, including among different foraging specialists (ecotypes) in sympatry. Given the strong matrifocal social structure of this species together with strong resource specializations, understanding the process of differentiation will require an understanding of the relative importance of both genetic drift and local adaptation. Here we provide a high-resolution analysis based on nuclear single-nucleotide polymorphic markers and inference about differentiation at both neutral loci and those potentially under selection. We find that all population comparisons, within or among foraging ecotypes, show significant differentiation, including populations in parapatry and sympatry. Loci putatively under selection show a different pattern of structure compared to neutral loci and are associated with gene ontology terms reflecting physiologically relevant functions (e.g. related to digestion). The pattern of differentiation for one ecotype in the North Pacific suggests local adaptation and shows some fixed differences among sympatric ecotypes. We suggest that differential habitat use and resource specializations have promoted sufficient isolation to allow differential evolution at neutral and functional loci, but that the process is recent and dependent on both selection and drift.

  20. [Selective-differential nutrient medium "Shewanella IRHLS agar" for isolation of Shewanella genus bacteria].

    Science.gov (United States)

    Sivolodsky, E P

    2015-01-01

    Development of a selective-differential nutrient medium for isolation of Shewanella genus bacteria. 73 strains of Shewanella bacteria (S. algae--3, S. baltica--26, S. putrefaciens--44) and 80 strains of 22 other bacteria genera were used. Shewanella species were identified by methods and criteria proposed by Nozue H. et al., 1992; Khashe S. et al., 1998. Nutrient media "Shewanella IRHLS Agar" for shewanella isolation was developed. Medium selective factors: irgazan DP-300 (I). 0.14-0.2 g/l and rifampicin (R) 0.0005-0.001 g/l. Shevanella colonies were detected by the production of hydrogen sulfide (H), lipase presence (L), lack of sorbitol fermentation (S). The medium suppressed the growth of hydrogen sulfide producers (Salmonella, Proteus) and blocked hydrogen sulfide production by Citrobacter. Growth of Escherichia, Enterobacter, Klebsiella, Shigella, Staphylococcus, Bacillus was also suppressed, Analytical sensitivity of the medium was 1-2 CFU/ml for Shewanella and Stenotrophomonas, Aerombnas, Serratia genera bacteria. 72 strains of Shewanella were isolated from water of Neva river in this medium, 91.7 ± 3.2% of those produced H2S. 1 strain of S. algae was isolated from clinical material. The developed media allows to use it in a complex for Stenotrophomo- nas sp., Aeromonas sp., Serratia sp., Citrobactersp. and Shewanella bacteria isolation.

  1. Isolation of UmRrm75, a gene involved in dimorphism and virulence of Ustilago maydis.

    Science.gov (United States)

    Rodríguez-Kessler, Margarita; Baeza-Montañez, Lourdes; García-Pedrajas, María D; Tapia-Moreno, Alejandro; Gold, Scott; Jiménez-Bremont, Juan F; Ruiz-Herrera, José

    2012-05-20

    Ustilago maydis displays dimorphic growth, alternating between a saprophytic haploid yeast form and a filamentous dikaryon, generated by mating of haploid cells and which is an obligate parasite. Induction of the dimorphic transition of haploid strains in vitro by change in ambient pH has been used to understand the mechanisms governing this differentiation process. In this study we used suppression subtractive hybridization to generate a cDNA library of U. maydis genes up-regulated in the filamentous form induced in vitro at acid pH. Expression analysis using quantitative RT-PCR showed that the induction of two unigenes identified in this library coincided with the establishment of filamentous growth in the acid pH medium. This expression pattern suggested that they were specifically associated to hyphal development rather than merely acid pH-induced genes. One of these genes, UmRrm75, encodes a protein containing three RNA recognition motifs and glycine-rich repeats and was selected for further study. The UmRrm75 gene contains 4 introns, and produces a splicing variant by a 3'-alternative splicing site within the third exon. Mutants deleted for UmRrm75 showed a slower growth rate than wild type strains in liquid and solid media, and their colonies showed a donut-like morphology on solid medium. Interestingly, although ΔUmRrm75 strains were not affected in filamentous growth induced by acid pH and oleic acid, they exhibited reduced mating, post-mating filamentous growth and virulence. Our data suggest that UmRrm75 is probably involved in cell growth, morphogenesis, and pathogenicity in U. maydis.

  2. A 'select and swap' strategy for the isolation of clones with tightly regulated transgenes.

    Science.gov (United States)

    Sullivan, M J; Carpenter, A J; Porter, A C

    2001-03-01

    Increasing numbers of biological problems are being addressed by genetic approaches that rely on inducible expression of transgenes. It is desirable that expression of such a transgene is tightly regulated, from close to zero expression in the 'off' state, to appreciable (at least physiological) expression in the 'on' state. Although there are many examples where tight regulation has been achieved, certain factors, including chromosomal position effects due to random integration of the transgene, often cause suboptimal inducibility and make the isolation of tightly regulated clones difficult and/or laborious. Here we describe a 'select and swap' strategy for the isolation, from a population of stable transfectants, of clones with tightly regulated transgenes. In this approach, a positively and negatively selectable, inducible marker gene is used to select for clones with optimal transgene regulation. After isolation of such clones, the marker gene is swapped with a linked gene of interest by the use of site-specific recombination. To test this strategy we introduced into human cells a plasmid with a tetracycline-inducible bacterial gpt gene linked to a promoterless luciferase gene, isolated clones with tight gpt expression and used the Cre/loxP site-specific recombination system to swap the gpt gene with the luciferase gene. We discuss ways for refining and developing the system and widening its applicability.

  3. Isolation and Characterization of Gut Bacterial Proteases Involved in Inducing Pathogenicity of Bacillus thuringiensis Toxin in Cotton Bollworm, Helicoverpa armigera

    Science.gov (United States)

    Regode, Visweshwar; Kuruba, Sreeramulu; Mohammad, Akbar S.; Sharma, Hari C.

    2016-01-01

    Bacillus thuringiensis toxin proteins are deployed in transgenic plants for pest management. The present studies were aimed at characterization of gut bacterial proteases involved in activation of inactive Cry1Ac protoxin (pro-Cry1Ac) to active toxin in Helicoverpa armigera. Bacterial strains were isolated from H. armigera midgut and screened for their proteolytic activation toward pro-Cry1Ac. Among 12 gut bacterial isolates seven isolates showed proteolytic activity, and proteases from three isolates (IVS1, IVS2, and IVS3) were found to be involved in the proteolytic conversion of pro-Cry1Ac into active toxin. The proteases from IVS1, IVS2, and IVS3 isolates were purified to 11.90-, 15.50-, and 17.20-fold, respectively. The optimum pH and temperature for gut bacterial protease activity was 8.0 and 40°C. Maximum inhibition of total proteolytic activity was exerted by phenylmethane sulfonyl fluoride followed by EDTA. Fluorescence zymography revealed that proteases from IVS1, IVS2, and IVS3 were chymotrypsin-like and showing protease band at ~15, 65, and 15 kDa, respectively. Active Cry1Ac formed from processing pro-Cry1Ac by gut bacterial proteases exhibited toxicity toward H. armigera. The gut bacterial isolates IVS1, IVS2, and IVS3 showed homology with B. thuringiensis (CP003763.1), Vibrio fischeri (CP000020.2), and Escherichia coli (CP011342.1), respectively. Proteases produced by midgut bacteria are involved in proteolytic processing of B. thuringiensis protoxin and play a major role in inducing pathogenicity of B. thuringiensis toxins in H. armigera. PMID:27766093

  4. Isolation and characterization of gut bacterial proteases involved in inducing pathogenicity of Bacillus thuringiensis toxin in cotton bollworm, Helicoverpa armigera

    Directory of Open Access Journals (Sweden)

    Visweshwar Regode

    2016-10-01

    Full Text Available Bacillus thuringiensis (Bt toxin proteins are deployed in transgenic plants for pest management. The present studies were aimed at characterization of gut bacterial proteases involved in activation of inactive Cry1Ac protoxin (pro-Cry1Ac to active toxin in Helicoverpa armigera. Bacterial strains were isolated from H. armigera midgut and screened for their proteolytic activation towards pro-Cry1Ac. Among twelve gut bacterial isolates seven isolates showed proteolytic activity, and proteases from three isolates (IVS1, IVS2 and IVS3 were found to be involved in the proteolytic conversion of pro-Cry1Ac into active toxin. The proteases from IVS1, IVS2 and IVS3 isolates were purified to 11.90-, 15.50- and 17.20-fold, respectively. The optimum pH and temperature for gut bacterial protease activity was 8.0 and 40 oC. Maximum inhibition of total proteolytic activity was exerted by PMSF followed by EDTA. Fluorescence zymography revealed that proteases from IVS1, IVS2, and IVS3 were chymotrypsin-like and showing protease band at ~15, 65 and 15 kDa, respectively. Active Cry1Ac formed from processing pro-Cry1Ac by gut bacterial proteases exhibited toxicity towards H. armigera. The gut bacterial isolates IVS1, IVS2 and IVS3 showed homology with Bacillus thuringiensis (CP003763.1, Vibrio fischeri (CP000020.2 and Escherichia coli (CP011342.1, respectively. Proteases produced by midgut bacteria are involved in proteolytic processing of Bt protoxin and play a major role in inducing pathogenicity of Bt toxins in H. armigera.

  5. Isolated Post-Transplantation Lymphoproliferative Disease Involving the Breast and Axilla as Peripheral T-cell Lymphoma

    Energy Technology Data Exchange (ETDEWEB)

    Hwang, Ji-Young [Department of Radiology, Kangnam Sacred Heart Hospital, Hallym University College of Medicine, Seoul 150-950 (Korea, Republic of); Cha, Eun Suk; Lee, Jee Eun [Department of Radiology, Ewha Womans University School of Medicine, Seoul 158-710 (Korea, Republic of); Sung, Sun Hee [Department of Pathology, Ewha Womans University School of Medicine, Seoul 158-710 (Korea, Republic of)

    2013-07-01

    Post-transplantation lymphoproliferative disorders (PTLDs) are a heterogeneous group of diseases that represent serious complications following immunosuppressive therapy for solid organ or hematopoietic-cell recipients. In contrast to B-cell PTLD, T-cell PTLD is less frequent and is not usually associated with Epstein Barr Virus infection. Moreover, to our knowledge, isolated T-cell PTLD involving the breast is extremely rare and this condition has never been reported previously in the literature. Herein, we report a rare case of isolated T-cell PTLD of the breast that occurred after a patient had been treated for allogeneic peripheral blood stem cell transplantation due to acute myeloblastic leukemia.

  6. SELECTION OF INDIGENOUS YEAST STRAINS ISOLATED FROM CV. GEWURZTRAMINER FROM ILOK WINEGROWING REGION

    Directory of Open Access Journals (Sweden)

    Dora DUJMOVIĆ

    2008-11-01

    Full Text Available A total of 36 indigenous strains belonging to the Saccharomyces sensu stricto complex isolated from the variety of Gewurztraminer from Ilok winegrowing region were tested with the purpose of making top-quality wines. Microfermentation was applied in order to examine the synthesis of ethyl alcohol and volatile acidity. Killer factor, synthesis of pectinase, ß-glucosidase and H2S was determined in isolated yeast strains. In the selection results, strains RO 1344, RO 1351, RO 1364 were the best evaluated and they should be examined in midifermentation.

  7. Learners Involvement in Materials Selection for Teaching English in Language Classroom at Aligarh Muslim University

    Directory of Open Access Journals (Sweden)

    Sheema Fatima

    2014-06-01

    Full Text Available The onset of the present paper throws light on materials selection and traditional outlook. Then the paper discusses four types of materials, more specifically, instructional, experiential, exploratory and elicitative materials and their use in language classroom. It discusses the role relationship between teachers, learners and materials in the present scenario at Aligarh Muslim University, keeping in consideration the requirements of the changing times. The paper aims to highlight the positive effects of learner’s involvement in Materials Selection process. The researcher has tried to analyze the findings on the basis of a questionnaire distributed among teachers and students at Undergraduate level. Some suggestions are recommended, on the basis of the findings from the data collected.

  8. Isolation of viable porcine islets by selective osmotic shock without enzymatic digestion.

    Science.gov (United States)

    Atwater, I; Guajardo, M; Caviedes, P; Jeffs, S; Parrau, D; Valencia, M; Romero, C; Arriagada, C; Caamaño, E; Salas, A; Olguin, F; Atlagich, M; Maas, R; Mears, D; Rojas, E

    2010-01-01

    Islet transplantation is a potential cure for type 1 diabetes, but clinical results have been disappointing. Currently, islet isolation is by enzymatic digestion of the pancreas which has significant pitfalls: warm ischemia exposure, collagenase-induced damage to the islet mass and viability, poor reproducibility, high cost, a relatively low number of islets obtained per whole pancreas, and selection of islets for collagenase resistance rather than for glucose responsiveness. In the present study we performed a series of experiments in a porcine model to demonstrate the feasibility of a new isolation method based on selective osmotic shock (SOS) using very high glucose solutions, doubling or tripling physiological osmotic strength. The SOS method can be carried out at room temperature or in the cold eliminating warm ischemia time which damages the islets. The SOS method does not depend on the texture of the pancreas so all pancreases can be processed identically and the process can be fully automated. The SOS method isolates all the islets of the pancreas regardless of size and shape allowing a greater number of islets to be harvested. The SOS method avoids exposure to toxins in collagenase solutions, is inexpensive and selects for islets with high concentrations of Glut 2 transporters, representing the best glucose responding islets. The SOS method showed a comparable recovery of islets from young pig pancreas and the islets showed improved viability. We conclude that the selective osmotic shock (SOS) method of separating islets from the pancreatic tissue is superior to the collagenase method.

  9. Engineering of Targeted Nanoparticles for Cancer Therapy Using Internalizing Aptamers Isolated by Cell-Uptake Selection

    Science.gov (United States)

    Xiao, Zeyu; Levy-Nissenbaum, Etgar; Alexis, Frank; Lupták, Andrej; Teply, Benjamin A.; Chan, Juliana M.; Shi, Jinjun; Digga, Elise; Cheng, Judy; Langer, Robert; Farokhzad, Omid C.

    2012-01-01

    One of the major challenges in the development of targeted nanoparticles (NPs) for cancer therapy is to discover targeting ligands that allow for differential binding and uptake by the target cancer cells. Using prostate cancer (PCa) as a model disease, we developed a cell-uptake selection strategy to isolate PCa-specific internalizing 2'-Omethyl RNA aptamers (Apts) for NP incorporation. Twelve cycles of selection and counter-selection were done to obtain a panel of internalizing Apts, which can distinguish PCa cells from non-prostate and normal prostate cells. After Apt characterization, size minimization, and conjugation of the Apts with fluorescently-labeled polymeric NPs, the NP-Apt bioconjugates exhibit PCa specificity and enhancement in cellular uptake when compared to non-targeted NPs lacking the internalizing Apts. Furthermore, when docetaxel, a chemotherapeutic agent used for the treatment of PCa, was encapsulated within the NP-Apt, a significant improvement in cytotoxicity was achieved in targeted PCa cells. Rather than isolating high-affinity Apts as reported in previous selection processes, our selection strategy was designed to enrich cancer-cell specific internalizing Apts. A similar cell-uptake selection strategy may be used to develop specific internalizing ligands for a myriad of other diseases and can potentially facilitate delivering various molecules, including drugs and siRNAs, into cells. PMID:22214176

  10. Isolation and Characterization of Frankia sp. Strain FaC1 Genes Involved in Nitrogen Fixation

    OpenAIRE

    Ligon, James M.; James P. Nakas

    1987-01-01

    Genomic DNA was isolated from Frankia sp. strain FaC1, an Alnus root nodule endophyte, and used to construct a genomic library in the cosmid vector pHC79. The genomic library was screened by in situ colony hybridization to identify clones of Frankia nitrogenase (nif) genes based on DNA sequence homology to structural nitrogenase genes from Klebsiella pneumoniae. Several Frankia nif clones were isolated, and hybridization with individual structural nitrogenase gene fragments (nifH, nifD, and n...

  11. Isolated bronchial artery involvement by polyarteritis nodosa presenting as hemoptysis: A case report

    Energy Technology Data Exchange (ETDEWEB)

    Park, Su Rin; Kwon, Jae Hyun; Lee, Kwang Hoon [Dongguk University Ilsan Hospital, Goyang (Korea, Republic of)

    2017-03-15

    Polyarteritis nodosa (PAN) is a systemic necrotizing vasculitis that involves medium- and small-sized arteries. PAN may affect any organ, and the presenting symptom of PAN varies depending on the organs affected. However, PAN generally spares the lung; thus, a report of PAN involving the bronchial artery is extremely rare, and hemoptysis has not been reported as the sole presenting symptom. Here, we report the case of a 39-year-old woman with hemoptysis who was diagnosed with PAN involving only the bronchial artery by angiography without involvement of the visceral arteries. Details of this case and a literature review are presented.

  12. SELECTIVE ISOLATION OF A GRAM NEGATIVE CARBAMATE PESTICIDE DEGRADING BACTERIUM FROM BRINJAL CULTIVATED SOIL

    Directory of Open Access Journals (Sweden)

    Dilara Islam Sharif

    2013-01-01

    Full Text Available A bacterial species having the ability to grow in the presence of carbosulfan pesticide “Marshall” was isolated from Solanum melongena (brinjal cultivated soil in Khulna region, Bangladeh, having a history of pesticide usage. The strain was morphologically and biochemically identified to belong to the genera Pseudomonas. A comparative study of growth of this strain with other isolated bacteria showed its ability to grow in the presence of different concentrations of Marshall. The susceptibility of the stain to Marshall was also assessed through disk diffusion assay which showed the strain to be resistant at concentrations of Marshal commonly used under field conditions. The selected strain also showed its capability to degrade Marshall through observed characteristics on sublimated agar plates. The biodegradation capability of the strain isolated in this study can be valuable for further study towards bioremediation of pesticide contaminated soils.

  13. Involvement of the MEN1 gene locus in familial isolated hyperparathyroidism

    DEFF Research Database (Denmark)

    Villablanca, Andrea; Wassif, Wassif S; Birket-Smith, Niels Thomas

    2002-01-01

    Familial isolated hyperparathyroidism (FIHP) is a hereditary disorder characterised by uni- or multiglandular parathyroid disease. A subset of families are likely to be genetic variants of other familial tumour syndromes in which PHPT is the main feature, for example multiple endocrine neoplasia...

  14. Molecular typing of Campylobacter jejuni isolates involved in a neonatal outbreak indicates nosocomial transmission

    DEFF Research Database (Denmark)

    Llovo, J.; Mateo, E.; Munoz, A.

    2003-01-01

    Genotypic typing by restriction fragment length polymorphism and pulsed-field gel electrophoresis showed that two neonates in a neonatal ward were infected with the same Campylobacter jejuni strain. Isolates from the mother and brother of the index patient were identical to each other but distinct...... from the neonatal type. Genotyping results therefore suggested that the neonatal C. jejuni infection was nosocomial in origin....

  15. Molecular phylogenetic and positive selection analysis of Japanese encephalitis virus strains isolated from pigs in China.

    Science.gov (United States)

    Liu, Wen-Jun; Zhu, Ming; Pei, Jing-Jing; Dong, Xiao-Ying; Liu, Wei; Zhao, Ming-Qiu; Wang, Jia-Ying; Gou, Hong-Chao; Luo, Yong-Wen; Chen, Jin-Ding

    2013-12-26

    Japanese encephalitis virus (JEV) is one of the most important virus which causes encephalitis. This disease is most prevalent in the south, southeast and the east region of Asia. In this study, two JEV strains, named JEV/SW/GD/01/2009 and JEV/SW/GZ/09/2004, were isolated from aborted fetuses and seminal fluid of pigs in China. To determine the characteristic of these virus isolates, the virulence of two newly JEV isolates was investigated, the result evidenced that the JEV/SW/GD/01/2009 did not kill mice, while the JEV/SW/GZ/09/2004 displayed neurovirulence with 0.925log10 p.f.u./LD50. Additionally, the full genome sequences of JEV were determined and compared with other known JEV strains. Results demonstrated that the genome of two JEV isolates was 10,976 nucleotides (nt) in length. As compared to the Chinese vaccine strain SA14-14-2, the JEV/SW/GD/01/2009 and the JEV/SW/GZ/09/2004 showed 99.7% and 97.5% identity at the nucleotide level, 99.6% and 96.7% identity at the amino acid level, respectively. Phylogenetic analysis, based on the full-length genome revealed that two JEV isolates were all clustered into genotype III compared to the reference strains. Furthermore, selection analyses revealed that dominant selective pressure acting on the JEV genome was purifying selection. Four sites under positive selection were identified: codon 521 (amino acid E-227), 2296 (amino acid NS4b-24), 3048 (amino acid NS5-521) and 3055 (amino acid NS5-528). Amino acid E-227 was proved to be related to neurovirulence. Taken together, the molecular epidemiology and functional of positively selected amino acid sites of two newly JEV isolates were fully understood, which might be helpful to predict possible changes in virulence. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. ISOLATION AND SELECTION OF THE NON TRANSDUCING E. COLI BACTERIOPHAGES FOR ANTI-COLIBACILLOSIS DRUGS

    Directory of Open Access Journals (Sweden)

    Skoblikov N. E.

    2016-10-01

    Full Text Available The isolation of E.coli phages from samples of natural and waste water obtained during expeditions in the different regions of Russian Federation was carried out. The obtained phages (286 isolates were tested for their ability to lyse the pathogenic strains of E. coli – pathogenic agents of pig colibacteriosis in Krasnodar region. The study was conducted of their ability to phage transduction, the molecular-genetic characterization and biotechnological parameters of selected bacteriophages. For first experimental design of veterinary drugs was selected 5 coliphages having no ability of plasmids transduction. It has been shown that all the investigated phages are representatives of T4-type phages of family Myoviridae. The reported study was partially supported by RFBR, research projects No. 08-04-99111, 09-04-10132, 16-44- 230855

  17. Laboratory identification of anaerobic bacteria isolated on Clostridium difficile selective medium.

    Science.gov (United States)

    Rodriguez, Cristina; Warszawski, Nathalie; Korsak, Nicolas; Taminiau, Bernard; Van Broeck, Johan; Delmée, Michel; Daube, Georges

    2016-06-01

    Despite increasing interest in the bacterium, the methodology for Clostridium difficile recovery has not yet been standardized. Cycloserine-cefoxitin fructose taurocholate (CCFT) has historically been the most used medium for C. difficile isolation from human, animal, environmental, and food samples, and presumptive identification is usually based on colony morphologies. However, CCFT is not totally selective. This study describes the recovery of 24 bacteria species belonging to 10 different genera other than C. difficile, present in the environment and foods of a retirement establishment that were not inhibited in the C. difficile selective medium. These findings provide insight for further environmental and food studies as well as for the isolation of C. difficile on supplemented CCFT.

  18. Selection and characterization of biofuel-producing environmental bacteria isolated from vegetable oil-rich wastes.

    Directory of Open Access Journals (Sweden)

    Almudena Escobar-Niño

    Full Text Available Fossil fuels are consumed so rapidly that it is expected that the planet resources will be soon exhausted. Therefore, it is imperative to develop alternative and inexpensive new technologies to produce sustainable fuels, for example biodiesel. In addition to hydrolytic and esterification reactions, lipases are capable of performing transesterification reactions useful for the production of biodiesel. However selection of the lipases capable of performing transesterification reactions is not easy and consequently very few biodiesel producing lipases are currently available. In this work we first isolated 1,016 lipolytic microorganisms by a qualitative plate assay. In a second step, lipolytic bacteria were analyzed using a colorimetric assay to detect the transesterification activity. Thirty of the initial lipolytic strains were selected for further characterization. Phylogenetic analysis revealed that 23 of the bacterial isolates were Gram negative and 7 were Gram positive, belonging to different clades. Biofuel production was analyzed and quantified by gas chromatography and revealed that 5 of the isolates produced biofuel with yields higher than 80% at benchtop scale. Chemical and viscosity analysis of the produced biofuel revealed that it differed from biodiesel. This bacterial-derived biofuel does not require any further downstream processing and it can be used directly in engines. The freeze-dried bacterial culture supernatants could be used at least five times for biofuel production without diminishing their activity. Therefore, these 5 isolates represent excellent candidates for testing biofuel production at industrial scale.

  19. Measurement of partition coefficients for selected polycyclic aromatic hydrocarbons between isolated plant cuticles and water.

    Science.gov (United States)

    Kim, Su-Jin; Lee, Hwang; Kwon, Jung-Hwan

    2014-10-01

    Partition coefficients between plant cuticles and water (Kcutw) were measured for 10 selected polycyclic aromatic hydrocarbons (PAHs) to evaluate the sorption capacity of plant cuticular layers for hydrophobic organic chemicals. The partitioning properties of PAHs between cuticles and water were evaluated by using (1) isolated cuticular layers and (2) leaf homogenate. The abaxial and adaxial cuticular layers of Euonymus japonicus were isolated by enzymatic digestion. A third-phase partitioning method using poly(dimethylsiloxane) (PDMS) was used to obtain Kcutw. The Kcutw values for the selected PAHs showed no significant differences between the abaxial and adaxial cuticular layers and ranged between 10(4.1) and 10(7.6). These values are close to or slightly higher than their 1-octanol/water partition coefficient (log Kow), indicating high sorption capacity of plant cuticles. On the contrary, partition coefficients between the lipid tissues of homogenized leaves and water were lower than those obtained using isolated cuticular layers by factors of 3.7-190, which is likely due to the breakdown of lipid layers. This indicates that the sorption of hydrophobic organic chemicals by plant leaves is better evaluated using isolated cuticles and that the sorption potential of plant leaves may be underestimated when leaf homogenates are used. Copyright © 2014 Elsevier B.V. All rights reserved.

  20. Selection and characterization of biofuel-producing environmental bacteria isolated from vegetable oil-rich wastes.

    Science.gov (United States)

    Escobar-Niño, Almudena; Luna, Carlos; Luna, Diego; Marcos, Ana T; Cánovas, David; Mellado, Encarnación

    2014-01-01

    Fossil fuels are consumed so rapidly that it is expected that the planet resources will be soon exhausted. Therefore, it is imperative to develop alternative and inexpensive new technologies to produce sustainable fuels, for example biodiesel. In addition to hydrolytic and esterification reactions, lipases are capable of performing transesterification reactions useful for the production of biodiesel. However selection of the lipases capable of performing transesterification reactions is not easy and consequently very few biodiesel producing lipases are currently available. In this work we first isolated 1,016 lipolytic microorganisms by a qualitative plate assay. In a second step, lipolytic bacteria were analyzed using a colorimetric assay to detect the transesterification activity. Thirty of the initial lipolytic strains were selected for further characterization. Phylogenetic analysis revealed that 23 of the bacterial isolates were Gram negative and 7 were Gram positive, belonging to different clades. Biofuel production was analyzed and quantified by gas chromatography and revealed that 5 of the isolates produced biofuel with yields higher than 80% at benchtop scale. Chemical and viscosity analysis of the produced biofuel revealed that it differed from biodiesel. This bacterial-derived biofuel does not require any further downstream processing and it can be used directly in engines. The freeze-dried bacterial culture supernatants could be used at least five times for biofuel production without diminishing their activity. Therefore, these 5 isolates represent excellent candidates for testing biofuel production at industrial scale.

  1. Consumer involvement in topic and outcome selection in the development of clinical practice guidelines.

    Science.gov (United States)

    Tong, Allison; Lopez-Vargas, Pamela; Howell, Martin; Phoon, Richard; Johnson, David; Campbell, Denise; Walker, Rowan G; Craig, Jonathan C

    2012-12-01

    Consumer involvement in guideline development is advocated, but minimal participation, such as a nominated consumer representative on a guideline working group, can inhibit their decision-making power and contribution. Little is known about how to involve consumers more effectively in guideline development. To describe a targeted approach for involving consumers actively in guideline development, by focusing on topic and outcome selection, and to discuss the impact on content and structure of the final guideline. Descriptive study. Patients and carers (n = 24) from a tertiary hospital in Sydney attended three structured peer-facilitated workshops to complete group-based exercises on topic and outcome selection for guidelines for early stage chronic kidney disease. These workshops were run in parallel with the guideline-writing group. For each exercise, participants formed small groups and facilitated their own discussion, recorded their responses and presented them to the wider group. The topics and outcomes identified were fed back to the guideline writers. The participants actively engaged in the workshop discussions and articulated topics and outcomes they perceived should be included in clinical guidelines. Four main changes to guideline-related outputs were observed. A new guideline subtopic was introduced, guidelines were consumer-endorsed, guideline recommendations and suggestions for clinical care were augmented with consumer-focused issues, and plain English guidelines were developed. Consumer workshops in parallel and feeding into guideline development can be a feasible and effective approach for active consumer contribution. This process can inform the development of both consumer-focused guidelines for clinicians and specific versions for consumers. © 2011 Blackwell Publishing Ltd.

  2. Resistance to Antibiotics, Biocides, Preservatives and Metals in Bacteria Isolated from Seafoods: Co-Selection of Strains Resistant or Tolerant to Different Classes of Compounds

    Science.gov (United States)

    Romero, José L.; Grande Burgos, María J.; Pérez-Pulido, Rubén; Gálvez, Antonio; Lucas, Rosario

    2017-01-01

    Multi-drug resistant bacteria (particularly those producing extended-spectrum β-lactamases) have become a major health concern. The continued exposure to antibiotics, biocides, chemical preservatives, and metals in different settings such as the food chain or in the environment may result in development of multiple resistance or co-resistance. The aim of the present study was to determine multiple resistances (biocides, antibiotics, chemical preservatives, phenolic compounds, and metals) in bacterial isolates from seafoods. A 75.86% of the 87 isolates studied were resistant to at least one antibiotic or one biocide, and 6.90% were multiply resistant to at least three biocides and at least three antibiotics. Significant (P isolates selected according to antimicrobial resistance profile and food type were identified by 16S rDNA sequencing and tested for copper and zinc tolerance. Then, the genetic determinants for biocide and metal tolerance and antibiotic resistance were investigated. The selected isolates were identified as Pseudomonas (63.33%), Acinetobacter (13.33%), Aeromonas (13.33%), Shewanella, Proteus and Listeria (one isolate each). Antibiotic resistance determinants detected included sul1 (43.33% of tested isolates), sul2 (6.66%), blaTEM (16.66%), blaCTX−M (16.66%), blaPSE (10.00%), blaIMP (3.33%), blaNDM−1 (3.33%), floR (16.66%), aadA1 (20.0%), and aac(6′)-Ib (16.66%). The only biocide resistance determinant detected among the selected isolates was qacEΔ1 (10.00%). A 23.30 of the selected isolates were able to grow on media containing 32 mM copper sulfate, and 46.60% on 8 mM zinc chloride. The metal resistance genes pcoA/copA, pcoR, and chrB were detected in 36.66, 6.66, and 13.33% of selected isolates, respectively. Twelve isolates tested positive for both metal and antibiotic resistance genes, including one isolate positive for the carbapenemase gene blaNDM−1 and for pcoA/copA. These results suggest that exposure to metals could co-select for

  3. DEVELOPMENT AND EVALUATION OF A SELECTIVE AND INDICATIVE MEDIUM FOR ISOLATION OF ACTINOBACILLUS-PLEUROPNEUMONIAE FROM TONSILS

    DEFF Research Database (Denmark)

    Jakobsen, Marianne; Nielsen, Jens

    1995-01-01

    In order to isolate ActinobacillIus pleuropneumoniae from mixed bacterial flora a selective and indicative medium was developed. The optimal concentrations of antibiotics were determined for selective chocolate agar (S-TSA) and selective blood agar (S-MBA) using a set of 25 strains of A. pleuropn......In order to isolate ActinobacillIus pleuropneumoniae from mixed bacterial flora a selective and indicative medium was developed. The optimal concentrations of antibiotics were determined for selective chocolate agar (S-TSA) and selective blood agar (S-MBA) using a set of 25 strains of A...

  4. The Micro-mechanism Involved and Wollastonite Signature in the Calcareous Precipitates of Marine Isolates.

    Science.gov (United States)

    Sarayu, K; Iyer, Nagesh R; Annaselvi, M; Ramachandra Murthy, A

    2016-03-01

    Micro-mechanical studies connecting the influence of extrinsic factors over intrinsic factors on 30 calcareous isolates obtained from marine sediment biofilms of the Bay of Bengal (Indian Ocean) revealed that the fate of calcareous crystal precipitation is highly dependent on factors like extracellular polysaccharides (EPS), organic carbon and nutrition. Further studies exemplified that EPS and the organic carbon secreted by the isolates controlled the dissemination of the calcareous crystals precipitated. From the study, it is evident that an EPS concentration of 7-15 mg l(-1) was found to enhance the dissemination of the calcareous crystals. Atomic force micrographs explain the nucleation behaviour and morphology of the calcareous crystals precipitated. X-ray diffraction (XRD) and energy-dispersive X-ray spectroscopy (EDAX) showed that the crystals were mainly composed of calcite and partially wollastonite.

  5. Prevalence, aetiology and antibiotic sensitivity profile of asymptomatic bacteriuria isolates from pregnant women in selected antenatal clinic from Nairobi, Kenya.

    Science.gov (United States)

    Ayoyi, Adelaide Ogutu; Kikuvi, Gideon; Bii, Christine; Kariuki, Samuel

    2017-01-01

    Asymptomatic bacteriuria (ASB) is the presence of bacteria in urine without apparent symptoms of urinary tract infections. The importance of asymptomatic bacteriuria lies in the insight it provides into symptomatic infections. To determine prevalence, bacterial isolates and Antibiotic Sensitivity Profile of asymptomatic bacterial urinary tract infection in pregnant women in selected clinics in Nairobi. This was a cross-sectional study involving women attending antenatal clinic at selected clinics of Nairobi County. The women who met the inclusion criteria were included in the study. The midstream urine samples of these women were subjected to microscopy, culture and sensitivity. A total of 1020 of women on their first antenatal clinic visit participated in the study; 219 of them had ASB, giving a prevalence of 21.5 % at 95% confidence level. Escherichia coli were the common organism isolated at 38.8%. The majority of the organisms were sensitive to imipenem and gentamycin. There is a high prevalence of ASB among pregnant women included in the study from the Nairobi county clinics. Therefore, routine ASB screening of pregnant women is recommended among the women attending antennal clinics in Nairobi county clinics.

  6. Tritium Suicide Selection Identifies Proteins Involved in the Uptake and Intracellular Transport of Sterols in Saccharomyces cerevisiae▿

    Science.gov (United States)

    Sullivan, David P.; Georgiev, Alexander; Menon, Anant K.

    2009-01-01

    Sterol transport between the plasma membrane (PM) and the endoplasmic reticulum (ER) occurs by a nonvesicular mechanism that is poorly understood. To identify proteins required for this process, we isolated Saccharomyces cerevisiae mutants with defects in sterol transport. We used Upc2-1 cells that have the ability to take up sterols under aerobic conditions and exploited the observation that intracellular accumulation of exogenously supplied [3H]cholesterol in the form of [3H]cholesteryl ester requires an intact PM-ER sterol transport pathway. Upc2-1 cells were mutagenized using a transposon library, incubated with [3H]cholesterol, and subjected to tritium suicide selection to isolate mutants with a decreased ability to accumulate [3H]cholesterol. Many of the mutants had defects in the expression and trafficking of Aus1 and Pdr11, PM-localized ABC transporters that are required for sterol uptake. Through characterization of one of the mutants, a new role was uncovered for the transcription factor Mot3 in controlling expression of Aus1 and Pdr11. A number of mutants had transposon insertions in the uncharacterized Ydr051c gene, which we now refer to as DET1 (decreased ergosterol transport). These mutants expressed Aus1 and Pdr11 normally but were severely defective in the ability to accumulate exogenously supplied cholesterol. The transport of newly synthesized sterols from the ER to the PM was also defective in det1Δ cells. These data indicate that the cytoplasmic protein encoded by DET1 is involved in intracellular sterol transport. PMID:19060182

  7. Compost as a source of microbial isolates for the bioremediation of heavy metals: In vitro selection

    Energy Technology Data Exchange (ETDEWEB)

    Vargas-Garcia, Maria del Carmen, E-mail: mcvargas@ual.es; Lopez, Maria Jose, E-mail: mllopez@ual.es; Suarez-Estrella, Francisca, E-mail: fsuarez@ual.es; Moreno, Joaquin, E-mail: jcasco@ual.es

    2012-08-01

    Heavy metal pollution has become a major environmental concern nowadays and the bioremediation of polluted habitats is an increasingly popular strategy due to both its efficiency and safety. A screening and selection protocol based on different composting processes was designed in order to isolate heavy metal-resistant microorganisms. A collection of 51 microorganisms was obtained and most of them showed the capability to tolerate heavy metals in multi-polluted aqueous systems (Cd(II), Cr(VI), Ni, Pb, Zn(II)), as well as to remove them. The highest detoxification ratios were observed for Pb. Some of the isolates detoxifying more than a 90% of this metal, while the other metals were removed in a range between 20% and 60%. The best isolates (Graphium putredinis, Fusarium solani, Fusarium sp. and Penicillium chrysogenum) were further assayed in order to determine the predominant removal mechanism and the potential use of their dead biomass as a biosorbent. Intracellular accumulation was the prevalent mechanism for most isolates and metals, with the exception of Ni. In this case, the proportion removed by extracellular adsorption was similar or even higher than that removed by intracellular accumulation. Thus, the efficiency of living cells was higher than that of dead biomass except in the case of Ni. - Highlights: Black-Right-Pointing-Pointer Composting is a good reservoir for the isolation of HM-resistant microorganisms. Black-Right-Pointing-Pointer Pb was the most removed heavy metal in multi-polluted aqueous systems. Black-Right-Pointing-Pointer Intracellular accumulation was the predominant mechanism for heavy metal removal. Black-Right-Pointing-Pointer Graphium putredinis, which detoxifies organic pollutants, was the most efficient isolate.

  8. Planning for selective amygdalohippocampectomy involving less neuronal ifber damage based on brain connectivity using tractography

    Institute of Scientific and Technical Information of China (English)

    Seung-Hak Lee; Mansu Kim; Hyunjin Park

    2015-01-01

    Temporal lobe resection is an important treatment option for epilepsy that involves removal of potentially essential brain regions. Selective amygdalohippocampectomy is a widely performed temporal lobe surgery. We suggest starting the incision for selective amygdalohippocampec-tomy at the inferior temporal gyrus based on diffusion magnetic resonance imaging (MRI) tractography. Diffusion MRI data from 20 normal participants were obtained from Parkinson’s Progression Markers Initiative (PPMI) database (www.ppmi-info.org). A tractography algorithm was applied to extract neuronal fiber information for the temporal lobe, hippocampus, and amygdala. Fiber information was analyzed in terms of the number of fibers and betweenness centrality. Distances between starting incisions and surgical target regions were also considered to explore the length of the surgical path. Middle temporal and superior temporal gyrus regions have higher connectivity values than the inferior temporal gyrus and thus are not good candi-dates for starting the incision. The distances between inferior temporal gyrus and surgical target regions were shorter than those between middle temporal gyrus and target regions. Thus, the in-ferior temporal gyrus is a good candidate for starting the incision. Starting the incision from the inferior temporal gyrus would spare the important (in terms of betweenness centrality values) middle region and shorten the distance to the target regions of the hippocampus and amygdala.

  9. A lateralized functional auditory network is involved in anuran sexual selection

    Indian Academy of Sciences (India)

    FEI XUE; GUANGZHAN FANG; XIZI YUE; ERMI ZHAO; STEVEN E BRAUTH; YEZHONG TANG

    2016-12-01

    Right ear advantage (REA) exists in many land vertebrates in which the right ear and left hemisphere preferentiallyprocess conspecific acoustic stimuli such as those related to sexual selection. Although ecological and neural mechanismsfor sexual selection have been widely studied, the brain networks involved are still poorly understood. In this study weused multi-channel electroencephalographic data in combination with Granger causal connectivity analysis to demonstrate,for the first time, that auditory neural network interconnecting the left and right midbrain and forebrain functionasymmetrically in the Emei music frog (Babina daunchina), an anuran species which exhibits REA. The results showedthe network was lateralized. Ascending connections between the mesencephalon and telencephalon were stronger in theleft side while descending ones were stronger in the right, which matched with the REA in this species and implied thatinhibition from the forebrainmay induce REA partly. Connections from the telencephalon to ipsilateral mesencephalon inresponse to white noise were the highest in the non-reproductive stage while those to advertisement calls were the highestin reproductive stage, implying the attention resources and living strategy shift when entered the reproductive season.Finally, these connection changes were sexually dimorphic, revealing sex differences in reproductive roles.

  10. Susceptibility of Mycosphaerella fijiensis isolates to the Higromicine B and Carbendazim selection agents

    Directory of Open Access Journals (Sweden)

    Mileidy Cruz-Martín

    2006-10-01

    Full Text Available The  study  of  Musa  -  Mycosphaerella  fijiensis  interactions  supported  by  molecular  tools  such  as  the  genetic transformation could contribute to elucidate new mechanisms related with the pathogenesis and through it to allow  the  development  of  new  selective  fungicides  as  well  as  the  elaboration  of  more  durable  resistance strategies.  This  work  had  as  objective,  to  determine  the  Minimum  Inhibitory  Concentration  (MIC  of  different antimicrobial substances commonly used as selection agents in the genetic transformation study of different species of Mycosphaerella (Hygromycin B and carbendazim. The agar dilution method was used and mycelial suspensions of nine of M. fijiensis isolated by several regions of Villa Clara and Ciego de Ávila was used as inoculums. Concentrations of these substances from 32.0 to 0.015 mg.l l -1  were rehearsed. It was possible to determine  the  MIC  of  both  antimicrobials  substances  against  the  M.fijiensis  isolated  using  this  method.  The MCI of the Hygromycin B for 100% of the isolated was less to 8 μg.ml l -1 and the rehearsed 90% of the isolated ones showed, for the carbendazim, MIC less or similar to 0.125 μg.ml l -1 . CCIBP-Pf-57 isolated did not achieve to be inhibited to the rehearsed concentrations of the Carbendazim. The results obtained in this work evidence the importance of knowing the MIC of the selective agents that are used in the transformation avoiding escapes in the selection. Key words: antimicrobial substance, genetic transformation, Minimal inhibitory concentration, mycelia suspension, Pseudosercospora  fijiensis

  11. Atypical scrapie isolates involve a uniform prion species with a complex molecular signature.

    Directory of Open Access Journals (Sweden)

    Dorothea R Götte

    Full Text Available The pathobiology of atypical scrapie, a prion disease affecting sheep and goats, is still poorly understood. In a previous study, we demonstrated that atypical scrapie affecting small ruminants in Switzerland differs in the neuroanatomical distribution of the pathological prion protein (PrP(d. To investigate whether these differences depend on host-related vs. pathogen-related factors, we transmitted atypical scrapie to transgenic mice over-expressing the ovine prion protein (tg338. The clinical, neuropathological, and molecular phenotype of tg338 mice is similar between mice carrying the Swiss atypical scrapie isolates and the Nor98, an atypical scrapie isolate from Norway. Together with published data, our results suggest that atypical scrapie is caused by a uniform type of prion, and that the observed phenotypic differences in small ruminants are likely host-dependant. Strikingly, by using a refined SDS-PAGE technique, we established that the prominent proteinase K-resistant prion protein fragment in atypical scrapie consists of two separate, unglycosylated peptides with molecular masses of roughly 5 and 8 kDa. These findings show similarities to those for other prion diseases in animals and humans, and lay the groundwork for future comparative research.

  12. Atypical scrapie isolates involve a uniform prion species with a complex molecular signature.

    Science.gov (United States)

    Götte, Dorothea R; Benestad, Sylvie L; Laude, Hubert; Zurbriggen, Andreas; Oevermann, Anna; Seuberlich, Torsten

    2011-01-01

    The pathobiology of atypical scrapie, a prion disease affecting sheep and goats, is still poorly understood. In a previous study, we demonstrated that atypical scrapie affecting small ruminants in Switzerland differs in the neuroanatomical distribution of the pathological prion protein (PrP(d)). To investigate whether these differences depend on host-related vs. pathogen-related factors, we transmitted atypical scrapie to transgenic mice over-expressing the ovine prion protein (tg338). The clinical, neuropathological, and molecular phenotype of tg338 mice is similar between mice carrying the Swiss atypical scrapie isolates and the Nor98, an atypical scrapie isolate from Norway. Together with published data, our results suggest that atypical scrapie is caused by a uniform type of prion, and that the observed phenotypic differences in small ruminants are likely host-dependant. Strikingly, by using a refined SDS-PAGE technique, we established that the prominent proteinase K-resistant prion protein fragment in atypical scrapie consists of two separate, unglycosylated peptides with molecular masses of roughly 5 and 8 kDa. These findings show similarities to those for other prion diseases in animals and humans, and lay the groundwork for future comparative research.

  13. Efficacy of Syringe Filtration for the Selective Isolation of Campylobacter from Chicken Carcass Rinse.

    Science.gov (United States)

    Chon, Jung-Whan; Kim, Hong-Seok; Kim, Dong-Hyeon; Kim, Young-Ji; Sung, Kidon; Kim, Hyunsook; Seo, Kun-Ho

    2017-06-01

    We investigated the efficacy of syringe filtration for selective isolation of Campylobacter from chicken carcass rinse by combining syringe filtration with the conventional culture method. Whole chicken carcass rinses were incubated in Bolton enrichment broth, set aside or subjected to syringe filtration, and streaked on Campy-Cefex agar with or without cefoperazone antibiotic supplement. Compared with the conventional method without filtration, 0.65-μm-pore-size syringe filtration resulted in a significantly higher number of Campylobacter-positive samples (23.8 to 37.5% versus 70.0 to 72.5%; P Campylobacter (93.8% versus 6.3 to 26.3%), and a lower growth index (1 = growth of a few colonies; 2 = growth of colonies on about half of the plate; and 3 = growth on most of the plate) for competing microbiota (2.9 to 3.0 versus 1.2 to 1.4). When syringe filtration was applied, agar plates containing the antibiotic had significantly less contamination (6.3% versus 26.3%; P Campylobacter isolation rate was similar (P > 0.05). Syringe filtration combined with conventional enrichment improved the rate and selectivity of Campylobacter isolation from chicken carcasses.

  14. Isolation and Fatty Acid Profile of Selected Microalgae Strains from the Red Sea for Biofuel Production

    Directory of Open Access Journals (Sweden)

    Khalid M. Abu-Salah

    2013-05-01

    Full Text Available The isolation of lipid-rich autochthonous strains of microalgae is a crucial stage for the development of a microalgae-based biofuel production plant, as these microalgae already have the necessary adaptations to withstand competition, predation and the temperatures observed at each production site. This is particularly important in extreme climates such as in Saudi Arabia. Resorting to fluorescence activated cell sorting (FACS we screened for and isolated several microalgal strains from samples collected from the Red Sea. Relying on the fluorescence of BODIPY 505/515 (4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diazasindacene and growth performance, four promising candidates were identified and the total lipid content and fatty acid profile was assessed for biofuels production. Selected isolates were classified as chlorophytes, belonging to three different genera: Picochlorum, Nannochloris and Desmochloris. The lipid contents were assessed microscopically by means of BODIPY 505/515-associated fluorescence to detect intracellular lipid bodies, which revealed several lipid drops in all selected strains. This result was confirmed by lipid gravimetric determination, which demonstrated that all strains under study presented inner cell lipid contents ranging from 20% to 25% of the biomass dry weight. Furthermore, the fatty acid methyl esters profile of all strains seems ideal for biodiesel production due to a low degree of polyunsaturated fatty acid methyl esters and high amount of palmitic and oleic acids.

  15. Virulence potential of Enterococcus gallinarum strains isolated from selected Nigerian traditional fermented foods

    Directory of Open Access Journals (Sweden)

    IYABO C. OLADIPO

    2014-08-01

    Full Text Available Five Enterococcus isolates from some Nigerian traditional fermented foods were identified as Enterococcus gallinarum by using phenotypic and genotypic tests. Safety properties such as antibiotic susceptibility, virulence gene detection, haemolysin, gelatinase and bacteriocin production were determined using standard methods. There was no resistance to clinically relevant antibiotics. Virulence gene for collagen binding antigen and aggregation substance were detected in 60% of the E. gallinarum strains; while surface adhesin was detected in 20%, but none of the strains had cytolysin activator and gelatinase. Phenotype characterizations of the E. gallinarum isolates indicated that none of the isolates produced haemolysin and gelatinase. Enterococcus gallinarum C103 and U82 had no antimicrobial activity against all the selected bacteria pathogens while E. gallinarum W184, T71 and W21 were active against some of the indicator bacteria pathogens. Only E. gallinarum T71 and W21 showed broad spectra of antimicrobial activity. Combination of virulence factors did not appear in these food isolates. Therefore, these strains particularly the two strains with high spectra of antimicrobial activity could be exploited as functional starters in foods.

  16. Compost as a source of microbial isolates for the bioremediation of heavy metals: in vitro selection.

    Science.gov (United States)

    Vargas-García, María del Carmen; López, María José; Suárez-Estrella, Francisca; Moreno, Joaquín

    2012-08-01

    Heavy metal pollution has become a major environmental concern nowadays and the bioremediation of polluted habitats is an increasingly popular strategy due to both its efficiency and safety. A screening and selection protocol based on different composting processes was designed in order to isolate heavy metal-resistant microorganisms. A collection of 51 microorganisms was obtained and most of them showed the capability to tolerate heavy metals in multi-polluted aqueous systems (Cd(II), Cr(VI), Ni, Pb, Zn(II)), as well as to remove them. The highest detoxification ratios were observed for Pb. Some of the isolates detoxifying more than a 90% of this metal, while the other metals were removed in a range between 20% and 60%. The best isolates (Graphium putredinis, Fusarium solani, Fusarium sp. and Penicillium chrysogenum) were further assayed in order to determine the predominant removal mechanism and the potential use of their dead biomass as a biosorbent. Intracellular accumulation was the prevalent mechanism for most isolates and metals, with the exception of Ni. In this case, the proportion removed by extracellular adsorption was similar or even higher than that removed by intracellular accumulation. Thus, the efficiency of living cells was higher than that of dead biomass except in the case of Ni.

  17. Isolation and Selection of Anti-Candida albicans Producing Lactic Acid Bacteria

    Directory of Open Access Journals (Sweden)

    Monthon LERTCANAWANICHAKUL

    2005-06-01

    Full Text Available The forty isolates of lactic acid bacteria (LAB were obtained from various fermented foods. The cross streak plate method was used to preliminary screen for antimicrobial activity. LAB were isolated by selective medium, Mann Rogosa Sharpe (MRS. Most of the isolates showed inhibition against Staphylococcus aureus TISTR 517, Bacillus subtilis TISTR 008, Micrococcus luteus TISTR 884, Escherichia coli TISTR 887, Pseudomonas aeruginosa TISTR 781, and Candida albicans DMST 5239. Only sterile culture supernatant of isolate No. L14, later identified as Lactococcus lactis, showed antifungal activity by means of agar well diffusion assay. The activity was stable during heat treatment and was retained even after autoclaving at 121 °C for 15 minutes. Maximum activity was observed at pH values between 2.5-4.0, and was lost at higher pH values. The anti-C. albicans activity was fully regained after readjustment of the pH to the initial value (pH 3.5.

  18. The genetic diversity and structure of 18 sheep breeds exposed to isolation and selection.

    Science.gov (United States)

    Ćurković, M; Ramljak, J; Ivanković, S; Mioč, B; Ivanković, A; Pavić, V; Brka, M; Veit-Kensch, C; Medugorac, I

    2016-02-01

    The phylogenetic layout of the genotyped (30 microsatellite) 18 sheep breeds in this study demands and provides the opportunity to evaluate both neutral and adaptive components of genetic diversity in a naturally and artificially selected and subdivided sheep population. Seven Pramenka strains from Bosnia and Herzegovina and Croatia characterized by a very low intensity of artificial selection, preserved the highest neutral genetic variability. Eight central and north-western European breeds under considerable artificial isolation and selection preserved the lowest genetic variability. Only combinations of various phylogenetic parameters offer a reasonable explanation for underlying evolutionary forces working in the investigated island and mainland sheep breeds under variable natural and artificial selection. More than 60% of total genetic, diversity was allocated to virtually unselected Pramenka strains, and an additional 25% to native moderately selected Graue Gehoernte Heidschnucke and intensively selected Ostfriesische Milchschafe. Some economically very important breeds and strains did not contribute to a pool with maximal genetic diversity, while they play an important role in the cultural heritage of respective countries.

  19. Selection of tomato plants resistant to a local Polish isolate of tomato spotted wilt virus (TSWV).

    Science.gov (United States)

    Czech, Andrzej S; Szklarczyk, Marek; Gajewski, Zbigniew; Zukowska, Ewa; Michalik, Barbara; Kobyłko, Tadeusz; Strzałka, Kazimierz

    2003-01-01

    We found that the Sw-5 gene confers resistance to one of the Polish isolates of tomato spotted wilt virus (TSWV). A series of tomato breeding accessions was analysed along with standards of resistance and susceptibility to TSWV. The presence of the Sw-5 gene was determined using the available PCR marker. Subsequently plants from these accessions were grown in the presence of the TSWV isolate from Poland. Some of them developed severe symptoms of the TSWV disease. Expression of the virus proteins was also assayed in tissues of the investigated plants. We found general agreement between either lack or presence of the disease symptoms, virus proteins and resistance gene. Some observed discrepancies of these data are also discussed. Our results indicate that marker-assisted selection can be used for breeding of the TSWV-resistant tomato in Poland.

  20. Isolation of a gene that is involved in Campylobacter jejuni 81116 cytotoxin activation.

    Science.gov (United States)

    Liu, Kaiyan; Fry, Benjamin N; Coloe, Peter J

    2007-02-01

    Cytotoxin fractions were isolated from Campylobacter jejuni 81116 and semi-purified by size-exclusion liquid chromatography. The fraction showing the strongest toxicity was injected into mice to produce antiserum. The antiserum was used to screen a C. jejuni 81116 cosmid library. Nine genes were identified in overlapping cosmid inserts that induced reactivity with the antiserum. One of these genes showed high similarity to a periplasmic protein of unknown function and its isogenic mutant showed decreased toxicity compared to the C. jejuni 81116 wild type. This gene contains a Gram-negative bacterial RTX toxin-activating protein C signature, which suggests it may play a role in C. jejuni 81116 cytotoxin activation.

  1. Potential of novel phytoecdysteroids isolated from Vitex doniana in the treatment depression: involvement of monoaminergic systems.

    Science.gov (United States)

    Ishola, Ismail O; Ochieng, Charles O; Olayemi, Sunday O; Jimoh, Memunat O; Lawal, Saheed M

    2014-12-01

    Vitex doniana Sweet (Verbanaceae) is used in traditional African medicine for the treatment of neurological disorders including depression. In our previous studies, three new phytoecdysteroids were isolated from methanol stem bark extract of V. doniana (VD) (11β-hydroxy-20-deoxyshidasterone, 21-hydroxyshidasterone, and 2,3-acetonide-24-hydroxyecdysone) along with known ecdysteroids. This study was designed to investigate antidepressant-like effect of VD and the isolated phytoecdysteroids in behavioral models of despair, forced-swim test (FST) and tail-suspension test (TST) in mice. VD (100 and 200mg/kg, p.o.) treatment reduced (Psulpiride (dopamine D2 receptor antagonist) prevented the antidepressant-like effect of 21-hydroxyshidasterone while the effects of 11β-hydroxy-20-deoxyshidasterone and 24-hydroxyecdysone were blocked by yohimbine or ketanserin in the FST. Moreover, the anti-immobility effect elicited by ajugasterone was prevented by prazosin (62.5μg/kg, i.p., α1-adrenoceptor antagonist) pretreatment. Our findings demonstrated that V. doniana and its phytoecdysteroids constituents elicited antidepressant-like effect in behavioral paradigm of despair. Furthermore, 21-hydroxyshidasterone produces its antidepressant-like effect through interaction with α2-adrenoceptor, 5-HT2A/2C receptor and dopamine D2-receptors but 11β-hydroxy-20-deoxyshidasterone and 24-hydroxyecdysone effects depend on interaction with α2-adrenoceptor and 5-HT2A/2C receptors while ajugasterone produces its action through interaction with post-synaptic α1-adrenoceptors. Thus, phytoecdysteroids could play a pivotal role in the treatment of major depression.

  2. Selection and geographic isolation influence hummingbird speciation: genetic, acoustic and morphological divergence in the wedge-tailed sabrewing (Campylopterus curvipennis

    Directory of Open Access Journals (Sweden)

    Ornelas Juan

    2011-02-01

    Full Text Available Abstract Background Mesoamerica is one of the most threatened biodiversity hotspots in the world, yet we are far from understanding the geologic history and the processes driving population divergence and speciation for most endemic taxa. In species with highly differentiated populations selective and/or neutral factors can induce rapid changes to traits involved in mate choice, promoting reproductive isolation between allopatric populations that can eventually lead to speciation. We present the results of genetic differentiation, and explore drift and selection effects in promoting acoustic and morphological divergence among populations of Campylopterus curvipennis, a lekking hummingbird with an extraordinary vocal variability across Mesoamerica. Results Analyses of two mitochondrial genes and ten microsatellite loci genotyped for 160 individuals revealed the presence of three lineages with no contemporary gene flow: C. c. curvipennis, C. c. excellens, and C. c. pampa disjunctly distributed in the Sierra Madre Oriental, the Tuxtlas region and the Yucatan Peninsula, respectively. Sequence mtDNA and microsatellite data were congruent with two diversification events: an old vicariance event at the Isthmus of Tehuantepec (c. 1.4 Ma, and a more recent Pleistocene split, isolating populations in the Tuxtlas region. Hummingbirds of the excellens group were larger, and those of the pampa group had shorter bills, and lineages that have been isolated the longest shared fewer syllables and differed in spectral and temporal traits of a shared syllable. Coalescent simulations showed that fixation of song types has occurred faster than expected under neutrality but the null hypothesis that morphological divergence resulted from drift was not rejected. Conclusions Our phylogeographic analyses uncovered the presence of three Mesoamerican wedge-tailed sabrewing lineages, which diverged at different time scales. These results highlight the importance of the

  3. Selective nitration of PsbO1 inhibits oxygen evolution from isolated Arabidopsis thylakoid membranes.

    Science.gov (United States)

    Takahashi, Misa; Shigeto, Jun; Sakamoto, Atsushi; Morikawa, Hiromichi

    2017-04-03

    Treatment of isolated Arabidopsis thaliana thylakoid membranes with nitrogen dioxide (NO2) induces selective nitration of the tyrosine residue at the ninth amino acid ((9)Tyr) of PsbO1. This selective nitration is triggered by light and is inhibited by photosynthetic electron transport inhibitors. Therefore, we postulated that, similar to (161)Tyr of D1 (YZ), (9)Tyr of PsbO1 is redox active and is selectively oxidized by photosynthetic electron transport in response to illumination to a tyrosyl radical that is highly susceptible to nitration. This tyrosyl radical may combine rapidly at diffusion-controlled rates with NO2 to form 3-nitrotyrosine. If this postulation is correct, the nitration of (9)Tyr of PsbO1 should decrease oxygen evolution activity. We investigated the effects of PsbO1 nitration on oxygen evolution from isolated thylakoid membranes, and found that nitration decreased oxygen evolution to ≥ 0% of the control. Oxygen evolution and nitration were significantly negatively correlated. This finding is consistent with redox active properties of the (9)Tyr gene of PsbO1, and suggests that PsbO1 (9)Tyr acts as an electron relay, such as YZ in the photosystem II oxygenic electron transport chain.

  4. Selection of candidate genes involved in the defense mechanisms of Phytophthora infestans against fungicides by EST analysis

    Science.gov (United States)

    Preliminary research using a functional genomics approach was conducted to gain insights on how P. infestans responds to fungicides and the possible implications of these responses on its ability to adapt to such selection pressure. Two isolates with subtle differences in mefenoxam resistance were e...

  5. [Isolation of peptide antibiotic virginiamycin components and selection of their producer Streptomyces virginiae].

    Science.gov (United States)

    Zvenigorodskiĭ, V I; Tiaglov, B V; Voeĭkova, T A

    2001-01-01

    A method for chromatographic separation and quantitative determination of individual components of the antibiotic virginiamycin, produced by microbiological synthesis (Streptomyces virginiae strain 147), is described. The components, M1-2 and S1-5, were isolated from fermentation broth and identified by HPTLC and HPLC (the results obtained using the two methods correlate well with each other). Conditions of culturing of the producer and compositions of nutritive media were optimized. Using UV irradiation as a mutagenic factor, the producer was selected for increased level of synthesis of the antibiotic; this was achieved by inducing mutations that impart resistance to virginiamycin and meta-fluorophenylalanine, an analog of phenylalanine.

  6. Involvement of the modifier gene of a human Mendelian disorder in a negative selection process.

    Directory of Open Access Journals (Sweden)

    Isabelle Jéru

    Full Text Available BACKGROUND: Identification of modifier genes and characterization of their effects represent major challenges in human genetics. SAA1 is one of the few modifiers identified in humans: this gene influences the risk of renal amyloidosis (RA in patients with familial Mediterranean fever (FMF, a Mendelian autoinflammatory disorder associated with mutations in MEFV. Indeed, the SAA1 alpha homozygous genotype and the p.Met694Val homozygous genotype at the MEFV locus are two main risk factors for RA. METHODOLOGY/PRINCIPAL FINDINGS: HERE, WE INVESTIGATED ARMENIAN FMF PATIENTS AND CONTROLS FROM TWO NEIGHBORING COUNTRIES: Armenia, where RA is frequent (24%, and Karabakh, where RA is rare (2.5%. Sequencing of MEFV revealed similar frequencies of p.Met694Val homozygotes in the two groups of patients. However, a major deficit of SAA1 alpha homozygotes was found among Karabakhian patients (4% as compared to Armenian patients (24% (p = 5.10(-5. Most importantly, we observed deviations from Hardy-Weinberg equilibrium (HWE in the two groups of patients, and unexpectedly, in opposite directions, whereas, in the two control populations, genotype distributions at this locus were similar and complied with (HWE. CONCLUSIONS/SIGNIFICANCE: The excess of SAA1alpha homozygotes among Armenian patients could be explained by the recruitment of patients with severe phenotypes. In contrast, a population-based study revealed that the deficit of alpha/alpha among Karabakhian patients would result from a negative selection against carriers of this genotype. This study, which provides new insights into the role of SAA1 in the pathophysiology of FMF, represents the first example of deviations from HWE and selection involving the modifier gene of a Mendelian disorder.

  7. Factors involved in selection of a career in surgery and orthopedics for medical students in Malawi.

    Science.gov (United States)

    Kollias, C; Banza, L; Mkandawire, N

    2010-03-01

    There is a critical shortage of Orthopedic Surgeons in Malawi as well as all countries in sub-Saharan Africa. To date, there is no published literature that has investigated surgical or Orthopedic career selection amongst African medical trainees. With the goal of facilitating recruitment into Surgery and Orthopedics in Malawi, we explored the key aspects of Malawian Medical Students' choice of careers in surgical disciplines. An on-line survey of all students in clinical years at the College of Medicine in Blantyre, Malawi was performed. The survey was anonymous and constructed de novo by a stringent process including Item Generation, Item reduction, Survey composition, Pre-testing, Assessment of Validity by a recognized survey expert, Pilot testing in on-line format by several Malawian Medical Students, and then formal survey testing. Surgery was the most popular specialty choice among the medical trainees (46%). General Surgery was the popular surgical specialty (27%), followed by Neurosurgery (22%) and Orthopedics (19%). The majority of students (67%) feared occupational exposure to HIV but this did not appear to be a factor in specialty choice (p = 0.9). Students with Orthopedic mentors were significantly more likely to choose Orthopedics as their first choice surgical specialty (p = 0.01). Despite limited resources and surgeons in sub-Saharan Africa, surgical specialties are desirable career choices. This is the first evaluation of factors involved in surgical or Orthopedic career selection in any African context. Future initiatives to improve exposure and mentorship in Orthopedics are fundamental to recruitment into the specialty.

  8. Selective vulnerability of the medullary thick ascending limb to anoxia in the isolated perfused rat kidney.

    OpenAIRE

    Brezis, M; Rosen, S.; Silva, P.; Epstein, F H

    1984-01-01

    A specific anatomical lesion sharply localized to the cells of the medullary thick ascending limbs (mTAL) and characterized by mitochondrial swelling progressing to nuclear pyknosis and cell death is elicited reproducibly in isolated rat kidneys perfused for 15 or 90 min with cell-free albumin-Ringer's medium gassed with 5% CO2, 95% O2 (O2 content, 1.5 vol/100 ml). The lesion, involving about half of mTALs, appears first in mTALs removed from vascular bundles and near the inner medulla, areas...

  9. Isolation and identification of Paenibacillus sp. FM-6, involved in the biotransformation of albendazole.

    Science.gov (United States)

    Jin, Lei; Zhang, Xiaojun; Sun, Xiumei; Shi, Hui; Li, Tiejun

    2014-10-01

    A strain, designated as FM-6, was isolated from fish. Based on the results of phenotypic, physiological characteristics, genotypic and phylogenetic analysis, strain FM-6 was finally identified as Paenibacillus sp. When albendazole was provided as the sole carbon source, strain FM-6 could grow and transform albendazole. About 82.7 % albendazole (50 mg/L) was transformed by strain FM-6 after 5 days incubation at 30 °C, 160 rpm. With HPLC-MS method, the transforming product of albendazole was researched. Based on the molecular weight and the retention time, product was identified as albendazole sulfoxide and the transforming pathway of albendazole by strain FM-6 was proposed finally. The optimum temperature and pH for the bacterium growth and albendazole transformation by strain FM-6 were both 30 °C and 7.0. Moreover, the optimum concentration of albendazole for the bacterium growth was 50 mg/L. Coupled with practical production, 50 mg/L was the optimum concentration of albendazole transformation for strain FM-6. This study highlights an important potential use of strain FM-6 for producing albendazole sulfoxide.

  10. Cardiac actions of phencyclidine in isolated guinea pig and rat heart: possible involvement of slow channels

    Energy Technology Data Exchange (ETDEWEB)

    Temma, K.; Akera, T.; Ng, Y.C.

    1985-03-01

    The mechanisms responsible for the positive inotropic effect of phencyclidine were studied in isolated preparations of guinea pig and rat heart. In electrically paced left atrial muscle preparations, phencyclidine increased the force of contraction; rat heart muscle preparations were more sensitive than guinea pig heart muscle preparations. The positive inotropic effect of phencyclidine was not significantly reduced by a combination of phentolamine and nadolol; however, the effect was competitively blocked by verapamil in the presence of phentolamine and nadolol. Inhibition of the outward K+ current by tetraethylammonium chloride also produced a positive inotropic effect; however, the effect of tetraethylammonium was reduced by phentolamine and nadolol, and was almost insensitive to verapamil. The inotropic effect of phencyclidine was associated with a marked prolongation of the action potential duration and a decrease in maximal upstroke velocity of the action potential, with no change in the resting membrane potential. The specific (/sup 3/H)phencyclidine binding observed with membrane preparations from guinea pig ventricular muscle was saturable with a single class of high-affinity binding site. This binding was inhibited by verapamil, diltiazem, or nitrendipine, but not by ryanodine or tetrodotoxin. These results suggest that the positive inotropic effect of phencyclidine results from enhanced Ca/sup 2 +/ influx via slow channels, either by stimulation of the channels or secondary to inhibition of outward K/sup +/ currents.

  11. IDENTIFICATION, ISOLATION AND AMPLIFICATION OF BRCA1 GENE INVOLVED IN BREAST CANCER

    Directory of Open Access Journals (Sweden)

    Karaneh Eftekhari

    2014-02-01

    Full Text Available Cancer is a disease that begins in the cells ofthe body which is characterized by uncontrolled, uncoordinated and undesirable cell division. If a cell accumulates critical mutations in five or six of the proto-oncogenes, tumour suppressor genes and DNA repair genes are likely to result in a fully malignant cell, capable of forming a tumour. In this work we described the isolation and amplification of the BRCA1 gene. Primers were designed and synthesised later used to amplify the BRCA1 gene. The total new workflow includes all steps from purified DNA to data analysis, and includes PCR for all amplicons covering the gene, PCR cleanup, cycle sequencing, electrophoresis, and data analysis. To simplify workflows and decrease the time-to-result, we focused on the method “one sample, one assay” approach. The success of this workflow was the 24-well plate design, which contained prespotted PCR primers covering the gene and also included multiplex nontemplate controls. The workflow was developed using a Genetic Analyzer and bands were observed.

  12. Comparison of subtypes of Listeria monocytogenes isolates from naturally contaminated watershed samples using a combination of non-selective and selective enrichment methods

    Science.gov (United States)

    Two enrichment methods for Listeria monocytogenes using Immuno Magnetic Separation were tested to determine if they selected the same subtypes of isolates. Both methods included a non-selective enrichment and one included subculture in Fraser Broth. Naturally contaminated watershed samples from the ...

  13. Characterization of Enterococcus isolates colonizing the intestinal tract of intensive care unit patients receiving selective digestive decontamination

    NARCIS (Netherlands)

    Bello Gonzalez, Teresita D.J.; Pham, Phu; Top, Janetta; Willems, Rob J.L.; Schaik, van Willem; Passel, van Mark W.J.; Smidt, Hauke

    2017-01-01

    Enterococci have emerged as important opportunistic pathogens in intensive care units (ICUs). In this study, enterococcal population size and Enterococcus isolates colonizing the intestinal tract of ICU patients receiving Selective Digestive Decontamination (SDD) were investigated. All nine

  14. Isolating quantum coherence with pathway-selective coherent multi-dimensional spectroscopy

    CERN Document Server

    Tollerud, Jonathan O; Davis, Jeffrey A

    2013-01-01

    Coherent coupling between spatially separated systems has long been explored as a necessary requirement for quantum information and cryptography. Recent discoveries suggest such phenomena appear in a much wider range of processes, including light-harvesting in photosynthesis. These discoveries have been facilitated by developments in coherent multi-dimensional spectroscopy (CMDS) that allow interactions between different electronic states to be identified in crowded spectra. For complex systems, however, spectral broadening and multiple overlapping peaks limit the ability to separate, identify and properly analyse all contributions. Here we demonstrate how pathway-selective CMDS can overcome these limitations to reveal, isolate and allow detailed analysis of weak coherent coupling between spatially separated excitons localised to different semiconductor quantum wells. Selective excitation of the coherence pathways, by spectrally shaping the laser pulses, provides access to previously hidden details and enable...

  15. SENSITIVITY OF MOLDS ISOLATED FROM WAREHOUSES OF FOOD PRODUCTION FACILITY ON SELECTED ESSENTIAL OILS

    Directory of Open Access Journals (Sweden)

    Łukasz Kręcidło

    2015-07-01

    Full Text Available Storage of raw materials is one of steps in food production chain. The aim of this study was to estimate the influence of selected essential oils on the growth of four fungal strains: Trichoderma viride, Rhizomucor miehei, Penicillium chrysogenum, Penicillium janthinellum. Strains were isolated from warehouses of the food production facility. Selected essential oils: thyme oil, rosewood oil and rosemary oil were used to assess antifungal activity. Chemical composition of essential oils was determined by Gas Chromatography-Mass Spectroscopy (GC-MS. Antifungal activity of essential oils was estimated in relative to peracetic acid (PAA and sterile water with Tween 80 (0,5%. The influence of essential oils on fungal growth was carried by medium poisoning method. Increment of fungal mycelium was measured every day by 10 days. The thyme essential oils totally inhibited fungal growth in the lowest concentration of 1 mm3·cm-3. The most resistant strain was Penicillium janthinellum.

  16. In vitro selection and characterization of putative probiotics isolated from the gut of Acipenser baerii (Brandt, 1869)

    NARCIS (Netherlands)

    Geraylou, Z.; Vanhove, M.P.M.; Souffreau, C.; Rurangwa, E.; Buyse, J.; Ollevier, F.

    2014-01-01

    To select and characterize potential probiotic bacteria from the gut microbiota of Siberian sturgeon (Acipenser baerii), 129 strains isolated from the hindgut were screened for antagonistic activity against five fish pathogens. Ten isolates showed antagonism towards three or more pathogens. Nine of

  17. Electro-membrane filtration for the selective isolation of bioactive peptides from an ?s2-casein hydrolysate

    NARCIS (Netherlands)

    Bargeman, Gerrald; Houwing, Joukje; Recio, Isidra; Koops, G.H.; van der Horst, Caroline

    2002-01-01

    For the isolation of the ingredients required for functional foods and nutraceuticals generally membrane filtration has too low a selectivity and chromatography is (too) expensive. Electro-membrane filtration (EMF) seems to be a breakthrough technology for the isolation of charged nutraceutical ingr

  18. Antimicrobial and antibiofilm effects of selected food preservatives against Salmonella spp. isolated from chicken samples.

    Science.gov (United States)

    Er, Buket; Demirhan, Burak; Onurdag, Fatma Kaynak; Ozgacar, Selda Özgen; Oktem, Aysel Bayhan

    2014-03-01

    Salmonella spp. are widespread foodborne pathogens that contaminate egg and poultry meats. Attachment, colonization, as well as biofilm formation capacity of Salmonella spp. on food and contact surfaces of food may cause continuous contamination. Biofilm may play a crucial role in the survival of salmonellae under unfavorable environmental conditions, such as in animal slaughterhouses and processing plants. This could serve as a reservoir compromising food safety and human health. Addition of antimicrobial preservatives extends shelf lives of food products, but even when products are supplemented with adequate amounts of preservatives, it is not always possible to inhibit the microorganisms in a biofilm community. In this study, our aims were i) to determine the minimum inhibitory concentrations (MIC) and minimum biofilm inhibitory concentrations (MBIC) of selected preservatives against planktonic and biofilm forms of Salmonella spp. isolated from chicken samples and Salmonella Typhimurium SL1344 standard strain, ii) to show the differences in the susceptibility patterns of same strains versus the planktonic and biofilm forms to the same preservative agent, and iii) to determine and compare antimicrobial and antibiofilm effects of selected food preservatives against Salmonella spp. For this purpose, Salmonella Typhimurium SL1344 standard strain and 4 Salmonella spp. strains isolated from chicken samples were used. Investigation of antimicrobial and antibiofilm effects of selected food preservatives against Salmonella spp. was done according to Clinical and Laboratory Standards Institute M100-S18 guidelines and BioTimer assay, respectively. As preservative agents, pure ciprofloxacin, sodium nitrite, potassium sorbate, sodium benzoate, methyl paraben, and propyl paraben were selected. As a result, it was determined that MBIC values are greater than the MIC values of the preservatives. This result verified the resistance seen in a biofilm community to food

  19. Isolation of the specific glomerular basement membrane antigen involved in Goodpasture syndrome.

    Science.gov (United States)

    Wieslander, J; Bygren, P; Heinegård, D

    1984-01-01

    The antigen involved in the glomerulonephritis associated with antibodies to glomerular basement membrane (GBM) was purified from human GBM digested with highly purified clostridial collagenase. The purified nonreduced sample contained two components with closely similar mobilities on sodium dodecyl sulfate/polyacrylamide gel electrophoresis. After reduction they moved as one, nonantigenic, component, corresponding to a molecular weight of 26,000. Immunologically identical aggregates of higher molecular weight (i.e., 48,000) were also identified in the crude digest. Reduction of such aggregates after purification released some protein with a molecular weight of 26,000, but a large proportion was insensitive to reduction. Seven patients with Goodpasture syndrome all had circulating anti-GBM antibodies directed only against the purified antigen. Images PMID:6324201

  20. Selective isolation of dematiaceous fungi from the workers of Atta laevigata (Formicidae: Attini).

    Science.gov (United States)

    Guedes, F L A; Attili-Angelis, D; Pagnocca, F C

    2012-01-01

    Leaf-cutting ants (Formicidae: Attini) are considered pests in agriculture for their impact in human crops, as they utilize leaf fragments to raise their fungal mutualist (Agaricales: Lepiotaceae). Basically, the basidiomycetous fungus is cultivated to supply food to adult workers and broads; in return, the ants protect it against natural enemies. However, recent studies have claimed that other microorganisms are associated to ant nests where a wide range of interactions may take place. To investigate the occurrence of dematiaceous fungi on the cuticle of Atta laevigata ants, 30 workers were sampled from an adult nest located in the surroundings of the Center for the Studies of Social Insects, UNESP-Rio Claro, SP, Brazil. The use of selective techniques to avoid high-sporulation fungi has been recommended and was tested in this study. To favor the isolation of the desired fungi, heads and cuticle scrapings of ant bodies were inoculated on Mycosel agar and incubated for 3 weeks at 35°C. Morphological and molecular methods were used to identify the filamentous fungi recovered. From 56 isolates, 19 were hyaline filamentous species, and among the remaining 37, some are mentioned as phyto-associated fungi like Alternaria arborescens, Bipolaris sorokiniana, Bipolaris eleusines, Bipolaris zeae, Curvularia trifolii, and Paraphaeosphaeria michotii. These species are reported from A. laevigata bodies for the first time. None of the isolation trials revealed the presence of the parasite Escovopsis or entomopathogenic fungi. The possible spread of the fungi in nature by the ants is discussed.

  1. Semi-selective culture medium for Exserohilum turcicum isolation from corn seeds

    Directory of Open Access Journals (Sweden)

    Roberto Luis De Rossi

    2014-06-01

    Full Text Available Northern corn leaf blight, caused by Exserohilum turcicum (Et, is a disease of widespread occurrence in regions where corn, sweetcorn and popcorn are grown. This disease has great potential to cause damage and has been studied for years, but the association of its causal agent with seeds remains unconfirmed. Thus, the availability of a sensitive method to detect and quantify the inoculum in seeds, even at low incidence, is essential. The aim of this study was to develop a method to detect and quantify the presence of the fungus infecting and infesting corn and popcorn seeds. Artificially and naturally infected seeds were employed to develop the medium. The semi-selective medium was composed of carbendazim (active ingredient (60 mg/L, captan (30 mg/L, streptomycin sulfate (500 mg/L and neomycin sulfate (600 mg/L aggregated to the medium lactose casein hydrolysate agar medium. By using this, Et was detected in naturally infected corn seeds, showing 0.124% incidence, in four out of ten analyzed samples. In addition, 1.04 conidia were detected per infested seed. By means of isolation, pathogenicity test, morphological characterization and comparison with descriptions of the species in the literature, the fungus isolated from the seeds was confirmed to be Et. Both infection and infestation were considered low; thus, for studies of Et detection in corn seeds, the use of semi-selective medium and more than 1,200 seeds/sample is suggested.

  2. Cluster Head Selection in a Homogeneous Wireless Sensor Network Ensuring Full Connectivity with Minimum Isolated Nodes

    Directory of Open Access Journals (Sweden)

    Tapan Kumar Jain

    2014-01-01

    Full Text Available The research work proposes a cluster head selection algorithm for a wireless sensor network. A node can be a cluster head if it is connected to at least one unique neighbor node where the unique neighbor is the one that is not connected to any other node. If there is no connected unique node then the CH is selected on the basis of residual energy and the number of neighbor nodes. With the increase in number of clusters, the processing energy of the network increases; hence, this algorithm proposes minimum number of clusters which further leads to increased network lifetime. The major novel contribution of the proposed work is an algorithm that ensures a completely connected network with minimum number of isolated nodes. An isolated node will remain only if it is not within the transmission range of any other node. With the maximum connectivity, the coverage of the network is automatically maximized. The superiority of the proposed design is verified by simulation results done in MATLAB, where it clearly depicts that the total numbers of rounds before the network dies out are maximum compared to other existing protocols.

  3. Isolation, Characterization, and Selection of Molds Associated to Fermented Black Table Olives

    Directory of Open Access Journals (Sweden)

    Simona L. Bavaro

    2017-07-01

    Full Text Available Table olives are one of the most important fermented food in the Mediterranean countries. Apart from lactic acid bacteria and yeasts that mainly conduct the olive fermentation, molds can develop on the brine surface, and can have either deleterious or useful effects on this process. From the food safety point of view, occurring molds could also produce mycotoxins, so, it is important to monitor and control them. In this respect, identification of molds associated to two Italian and two Greek fermented black table olives cultivars, was carried out. Sixty strains were isolated and molecularly identified as Penicillium crustosum (21, P. roqueforti (29, P. paneum (1, P. expansum (6, P. polonicum (2, P. commune (1. A group of 20 selected isolates was subjected to technological (beta-glucosidase, cellulolytic, ligninolytic, pectolytic, and xylanolytic activities; proteolytic enzymes and safety (biogenic amines and secondary metabolites, including mycotoxins characterization. Combining both technological (presence of desired and absence of undesired enzymatic activities and safety aspects (no or low production of biogenic amines and regulated mycotoxins, it was possible to select six strains with biotechnological interest. These are putative candidates for future studies as autochthonous co-starters with yeasts and lactic acid bacteria for black table olive production.

  4. Isolation and characterization of maize PMP3 genes involved in salt stress tolerance.

    Directory of Open Access Journals (Sweden)

    Jing Fu

    Full Text Available Plasma membrane protein 3 (PMP3, a class of small hydrophobic polypeptides with high sequence similarity, is responsible for salt, drought, cold, and abscisic acid. These small hydrophobic ploypeptides play important roles in maintenance of ion homeostasis. In this study, eight ZmPMP3 genes were cloned from maize and responsive to salt, drought, cold and abscisic acid. The eight ZmPMP3s were membrane proteins and their sequences in trans-membrane regions were highly conserved. Phylogenetic analysis showed that they were categorized into three groups. All members of group II were responsive to ABA. Functional complementation showed that with the exception of ZmPMP3-6, all were capable of maintaining membrane potential, which in turn allows for regulation of intracellular ion homeostasis. This process was independent of the presence of Ca(2+. Lastly, over-expression of ZmPMP3-1 enhanced growth of transgenic Arabidopsis under salt condition. Through expression analysis of deduced downstream genes in transgenic plants, expression levels of three ion transporter genes and four important antioxidant genes in ROS scavenging system were increased significantly in transgenic plants during salt stress. This tolerance was likely achieved through diminishing oxidative stress due to the possibility of ZmPMP3-1's involvement in regulation of ion homeostasis, and suggests that the modulation of these conserved small hydrophobic polypeptides could be an effective way to improve salt tolerance in plants.

  5. The role of the catalysts with highly dispersed and isolated active sites in the selective oxidation of light hydrocarbons

    Institute of Scientific and Technical Information of China (English)

    WANG Hongxuan; ZHAO Zhen

    2005-01-01

    This review summarizes the role of catalysts with highly dispersed and isolated active sites (active sites: supported atoms f≤0.5 % ) in the selective oxidation of light hydrocarbons, such as methane, ethane and propane, into oxygenatesand the epoxidation of olefins. The plausible structures of the highly dispersed and isolated active species, as well as their effects on the catalytic performances are discussed. The special physico-chemical properties and the functional mechanism of the catalysts with highly dispersed and isolated active sites, as well as the preparation, characterization of the catalysts with highly dispersed and isolated active sites and their applications in other types of reactions of lower hydrocarbons are summarized.

  6. Graphene oxide-rare earth metal-organic framework composites for the selective isolation of hemoglobin.

    Science.gov (United States)

    Liu, Jia-Wei; Zhang, Yue; Chen, Xu-Wei; Wang, Jian-Hua

    2014-07-09

    Graphene oxide-La(BTC)(H2O)6 (H3BTC=1,3,5-benzenetricarboxylic acid) metal organic framework composites (LaMOF-GOn, n = 1-6, corresponding to the percentage of GO at 1, 2, 3, 4, 5, and 10%) are prepared through a simple and large-scale method at room temperature. The obtained composites are characterized by ATR-FTIR spectra, SEM, XRD, TGA, and N2 adsorption-desorption isotherm. The presence of GO significantly changes the morphologies of the composites from spindly rectangular rods to irregular thick blocks and increases their surface area from 14.8 cm(2) g(-1) (LaMOFs) to 26.6 cm(2) g(-1) (LaMOF-GO3), whereas at the same time, the crystalline structure of La(BTC)(H2O)6 is maintained. As a novel solid-phase adsorbent the LaMOF-GO composite exhibits outstanding adsorption properties for proteins. The strong hydrophobic interaction, especially π-π interaction between protein and the composite, is the main driving force for protein adsorption. In particular, highly selective isolation of hemoglobin (Hb) is achieved by using LaMOF-GO3 composite as sorbent in 4 mM B-R buffer containing 0.05 mol L(-1) NaCl at pH 8. The retained Hb could be effectively recovered with a 1 mM B-R buffer at pH 10, giving rise to a recovery of 63%. The practical applicability of the LaMOF-GO3 composite is demonstrated by the selective adsorption of Hb from human whole blood, and SDS-PAGE assays indicate that Hb could be selectively isolated with high purity from biological samples of complex matrixes.

  7. A Bacillus thuringiensis isolation method utilizing a novel stain, low selection and high throughput produced atypical results

    Directory of Open Access Journals (Sweden)

    Ammons David

    2005-09-01

    Full Text Available Abstract Background Bacillus thuringiensis is a bacterium known for producing protein crystals with insecticidal properties. These toxins are widely sought after for controlling agricultural pests due to both their specificity and their applicability in transgenic plants. There is great interest in isolating strains with improved or novel toxin characteristics, however isolating B. thuringiensis from the environment is time consuming and yields relatively few isolates of interest. New approaches to B. thuringiensis isolation have been, and continue to be sought. In this report, candidate B. thuringiensis isolates were recovered from environmental samples using a combination of a novel stain, high throughput and reduced selection. Isolates were further characterized by SDS-PAGE, light microscopy, PCR, probe hybridization, and with selected isolates, DNA sequencing, bioassay or Electron Microscopy. Results Based on SDS-PAGE patterns and the presence of cry genes or a crystal, 79 candidate, non-clonal isolates of B. thuringiensis were identified from 84 samples and over 10,000 colonies. Although only 16/79 (20% of the isolates showed DNA homology by Probe Hybridization or PCR to common cry genes, initial characterization revealed a surprisingly rich library that included a putative nematocidal gene, a novel filamentous structure associated with a crystal, a spore with spikes originating from a very small parasporal body and isolates with unusually small crystals. When compared to reports of other screens, this screen was also atypical in that only 3/79 isolates (3.8% produced a bipyramidal crystal and 24/79 (30% of the isolates' spores possessed an attached, dark-staining body. Conclusion Results suggest that the screening methodology adopted in this study might deliver a vastly richer and potentially more useful library of B. thuringiensis isolates as compared to that obtained with commonly reported methodologies, and that by extension

  8. Parental Involvement in Selected PISA Countries and Economies. OECD Education Working Papers, No. 73

    Science.gov (United States)

    Borgonovi, Francesca; Montt, Guillermo

    2012-01-01

    Studies have highlighted the beneficial effects of parental involvement in children's educational lives. Few studies, however, analyse parental involvement in a cross-national perspective and few evaluate a wide array of forms of involvement. In 2009, 14 countries and economies implemented the parental questionnaire option in the PISA 2009 cycle.…

  9. Isolation of deoxynivalenol-transforming bacteria from the chicken intestines using the approach of PCR-DGGE guided microbial selection

    Directory of Open Access Journals (Sweden)

    Li Xiu-Zhen

    2010-06-01

    Full Text Available Abstract Background Contamination of grains with trichothecene mycotoxins, especially deoxynivalenol (DON, has been an ongoing problem for Canada and many other countries. Mycotoxin contamination creates food safety risks, reduces grain market values, threatens livestock industries, and limits agricultural produce exports. DON is a secondary metabolite produced by some Fusarium species of fungi. To date, there is a lack of effective and economical methods to significantly reduce the levels of trichothecene mycotoxins in food and feed, including the efforts to breed Fusarium pathogen-resistant crops and chemical/physical treatments to remove the mycotoxins. Biological approaches, such as the use of microorganisms to convert the toxins to non- or less toxic compounds, have become a preferred choice recently due to their high specificity, efficacy, and environmental soundness. However, such approaches are often limited by the availability of microbial agents with the ability to detoxify the mycotoxins. In the present study, an approach with PCR-DGGE guided microbial selection was developed and used to isolate DON -transforming bacteria from chicken intestines, which resulted in the successful isolation of several bacterial isolates that demonstrated the function to transform DON to its de-epoxy form, deepoxy-4-deoxynivalenol (DOM-1, a product much less toxic than DON. Results The use of conventional microbiological selection strategies guided by PCR-DGGE (denaturing gradient gel electrophoresis bacterial profiles for isolating DON-transforming bacteria has significantly increased the efficiency of the bacterial selection. Ten isolates were identified and isolated from chicken intestines. They were all able to transform DON to DOM-1. Most isolates were potent in transforming DON and the activity was stable during subculturing. Sequence data of partial 16S rRNA genes indicate that the ten isolates belong to four different bacterial groups

  10. Highly selective isolation and purification of heme proteins in biological samples using multifunctional magnetic nanospheres.

    Science.gov (United States)

    Liu, Yating; Li, Yan; Wei, Yun

    2014-12-01

    Magnetic particles with suitable surface modification are capable of binding proteins selectively, and magnetic separations have advantages of rapidity, convenience, and high selectivity. In this paper, new magnetic nanoparticles modified with imidazolium ionic liquid (Fe3O4 @SiO2 @ILs) were successfully fabricated. N-Methylimidazolium was immobilized onto silica-coated magnetic nanoparticles via γ-chloropropyl modification as a magnetic nanoadsorbent for heme protein separation. The particle size was about 90 nm without significant aggregation during the preparation process. Hemoglobin as one of heme proteins used in this experiment was compared with other nonheme proteins. It has been found that the magnetic nanoparticles can be used for more rapid, efficient, and specific adsorption of hemoglobin with a binding capacity as high as 5.78 mg/mg. In comparison with other adsorption materials of proteins in the previous reports, Fe3 O4 @SiO2 @ILs magnetic nanoparticles exhibit the excellent performance in isolation of heme proteins with higher binding capacity and selectivity. In addition, a short separation time makes the functionalized nanoparticles suitable for purifying unstable proteins, as well as having other potential applications in a variety of biomedical fields.

  11. A new selective medium for isolation of Clavibacter michiganensis subsp. michiganensis from tomato plants and seed.

    Science.gov (United States)

    Ftayeh, Radwan M; von Tiedemann, Andreas; Rudolph, Klaus W E

    2011-11-01

    A new selective and highly sensitive medium was developed for isolation of Clavibacter michiganensis subsp. michiganensis (Cmm), the causal agent of bacterial canker of tomato, from seed and latently infected plants. The new medium (BCT) proved to be superior to all published semiselective media for Cmm and is denoted as selective medium because of (i) its mean plating efficiency, amounting to ≤89% within 7 days for all 30 Cmm strains from different sources tested; (ii) the high selectivity, because accompanying bacterial species occurring on tomato plants and seed or bacteria obtained from culture collections were inhibited to an extent of 98 to 100%; and (iii) the remarkable detection sensitivity. Thus, 8 CFU of Cmm in field plant homogenates containing 12,750 CFU of accompanying saprophytes were detected on BCT. Under these extreme conditions, all of the published semiselective media (D2, KBT, D2ANX, SCM, mSCM, CMM1, mCNS, and EPPO) gave false-negative results. Either some media were rather toxic and Cmm growth was also inhibited or the other, less toxic media allowed growth of high numbers of saprophytes, so that Cmm growth was suppressed. Exclusively, BCT also supported growth of the closely related C. michiganensis subsp. insidiosus, nebraskensis, and tessellarius. The new medium is recommended for Cmm detection in tomato seed, and in symptomless tomato plantlets, to improve disease control of bacterial canker of tomato.

  12. Comparative antimicrobial efficacy of selected root canal irrigants on commonly isolated microorganisms in endodontic infection.

    Science.gov (United States)

    Dubey, Sandeep; Saha, Suparna Ganguly; Rajkumar, Balakrishnan; Dhole, Tapan Kumar

    2017-01-01

    This study aims to evaluate and compare the antimicrobial efficacy of three selected root canal irrigants (BioPure MTAD, metronidazole, aztreonam) against microbes commonly isolated from polymicrobial microbiota of root canal infection. This study was designed with four experimental groups (Group I - Bacteroides fragilis, Group II - Propionibacterium acnes, Group III - Enterococcus faecalis, Group IV - Candida albicans) based on the microbes selected for the study. Group I and Group II bacteria were used to compare and evaluate antimicrobial effect of BioPure MTAD, metronidazole, aztreonam, and normal saline. Group III and Group IV bacteria were used to compare and evaluate antimicrobial efficacy of BioPure MTAD, aztreonam, and normal saline. Normal saline was used as a control irrigant in this study. Agar disc diffusion method was applied to assess and compare the antimicrobial action of selected irrigants. Metronidazole was found to be the most effective root canal irrigant against B. fragilis and P. acnes among the tested irrigants. Mean zone of inhibition against E. faecalis has been shown to be maximum by BioPure MTAD, followed by aztreonam. Antifungal effect against C. albicans was only shown by BioPure MTAD. Overall, BioPure MTAD is the most effective root canal irrigant as it has shown an antibacterial effect against all the tested microorganisms. However, metronidazole showed maximum antibacterial effect against obligate anaerobes. Aztreonam also showed an antibacterial effect in the present study, raising its possibility to be used as a root canal irrigant in the future.

  13. Selection of isolated and individual single-walled carbon nanotube from a film and its usage in nanodevices

    Institute of Scientific and Technical Information of China (English)

    Wang Gong-Tang

    2011-01-01

    Individual and isolated single-walled carbon nanotubes (SWNTs) are important for fabricating relevant nanodevices and studying the properties of the SWNT devices. In this work, we demonstrate that individual and isolated SWNT can be selected and obtained from a film containing a huge number of SWNTs. By using both the polymethylmethacrylate (PMMA) as a negative resist and the electron beam lithography, the selected SWNT can be fixed on a substrate, while the other SWNTs in the film can lift off. The selected SWNT can be used to fabricate nanodevice and a gas sensor of oxygen is demonstrated in this work.

  14. Selection of oleuropein-degrading lactic acid bacteria strains isolated from fermenting Moroccan green olives

    Directory of Open Access Journals (Sweden)

    Ghabbour, N.

    2011-03-01

    Full Text Available A total of 177 strains of lactic acid bacteria (LAB were isolated from early-stage Moroccan Picholine green olive fermentation, including Lactobacillus plantarum (44.63%, Lactobacillus pentosus (25.99%, Lactobacillus brevis (9.61% and Pediococcus pentosaceus (19.77%. All the isolates were screened for their tolerance to olive leaf extract and oleuropein. Most of the isolates (85.3% were found able to degrade oleuropein, when evaluated by either oleuropein or 5-Bromo- 4-chloro-3-indolyl β-D-glucuronide (X-Gluc as substrates. The biodegradation capacity of the selected strains of each species was confirmed by HPLC analysis.

    Un total de 177 cepas de bacterias ácido lácticas (LAB fueron aisladas en las primeras etapas de la fermentación de aceitunas verdes marroquíes Picholine, incluyendo Lactobacillus plantarum (44.63%, Lactobacillus pentosus (25.99%, Lactobacillus brevis (9.61% y Pediococcus pentosaceus (19.77%. Todos los aislados fueron evaluados mediante su tolerancia a extractos de hojas de olivo y oleuropeína. La mayoría de los aislados (85,3% degradaron oleuropeína, cuando fueron evaluados usando oleuropeína o 5-Bromo-4-cloro- 3-indolil β-D-glucuronido (X-Gluc como sustrato. La capacidad de biodegradación de las cepas seleccionadas para cada especie fue confirmada mediante análisis por HPLC.

  15. Involvement of oxidative stress and mitochondrial/lysosomal cross-talk in olanzapine cytotoxicity in freshly isolated rat hepatocytes.

    Science.gov (United States)

    Eftekhari, Aziz; Azarmi, Yadollah; Parvizpur, Alireza; Eghbal, Mohammad Ali

    2016-01-01

    1. Olanzapine (OLZ) is a widely used atypical antipsychotic agent for the treatment of schizophrenia and other disorders. Serious hepatotoxicity and elevated liver enzymes have been reported in patients receiving OLZ. However, the cellular and molecular mechanisms of the OLZ hepatotoxicity are unknown. 2. In this study, the cytotoxic effect of OLZ on freshly isolated rat hepatocytes was assessed. Our results showed that the cytotoxicity of OLZ in hepatocytes is mediated by overproduction of reactive oxygen species (ROS), mitochondrial potential collapse, lysosomal membrane leakiness, GSH depletion and lipid peroxidation preceding cell lysis. All the aforementioned OLZ-induced cellular events were significantly (p lysosomal involvement following the initiation of oxidative stress in hepatocytes.

  16. FAS-Based Cell Depletion Facilitates the Selective Isolation of Mouse Induced Pluripotent Stem Cells

    Science.gov (United States)

    Warlich, Eva; Schambach, Axel; Lock, Dominik; Wedekind, Dirk; Glage, Silke; Eckardt, Dominik; Bosio, Andreas; Knöbel, Sebastian

    2014-01-01

    Cellular reprogramming of somatic cells into induced pluripotent stem cells (iPSC) opens up new avenues for basic research and regenerative medicine. However, the low efficiency of the procedure remains a major limitation. To identify iPSC, many studies to date relied on the activation of pluripotency-associated transcription factors. Such strategies are either retrospective or depend on genetically modified reporter cells. We aimed at identifying naturally occurring surface proteins in a systematic approach, focusing on antibody-targeted markers to enable live-cell identification and selective isolation. We tested 170 antibodies for differential expression between mouse embryonic fibroblasts (MEF) and mouse pluripotent stem cells (PSC). Differentially expressed markers were evaluated for their ability to identify and isolate iPSC in reprogramming cultures. Epithelial cell adhesion molecule (EPCAM) and stage-specific embryonic antigen 1 (SSEA1) were upregulated early during reprogramming and enabled enrichment of OCT4 expressing cells by magnetic cell sorting. Downregulation of somatic marker FAS was equally suitable to enrich OCT4 expressing cells, which has not been described so far. Furthermore, FAS downregulation correlated with viral transgene silencing. Finally, using the marker SSEA-1 we exemplified that magnetic separation enables the establishment of bona fide iPSC and propose strategies to enrich iPSC from a variety of human source tissues. PMID:25029550

  17. FAS-based cell depletion facilitates the selective isolation of mouse induced pluripotent stem cells.

    Directory of Open Access Journals (Sweden)

    Eva Warlich

    Full Text Available Cellular reprogramming of somatic cells into induced pluripotent stem cells (iPSC opens up new avenues for basic research and regenerative medicine. However, the low efficiency of the procedure remains a major limitation. To identify iPSC, many studies to date relied on the activation of pluripotency-associated transcription factors. Such strategies are either retrospective or depend on genetically modified reporter cells. We aimed at identifying naturally occurring surface proteins in a systematic approach, focusing on antibody-targeted markers to enable live-cell identification and selective isolation. We tested 170 antibodies for differential expression between mouse embryonic fibroblasts (MEF and mouse pluripotent stem cells (PSC. Differentially expressed markers were evaluated for their ability to identify and isolate iPSC in reprogramming cultures. Epithelial cell adhesion molecule (EPCAM and stage-specific embryonic antigen 1 (SSEA1 were upregulated early during reprogramming and enabled enrichment of OCT4 expressing cells by magnetic cell sorting. Downregulation of somatic marker FAS was equally suitable to enrich OCT4 expressing cells, which has not been described so far. Furthermore, FAS downregulation correlated with viral transgene silencing. Finally, using the marker SSEA-1 we exemplified that magnetic separation enables the establishment of bona fide iPSC and propose strategies to enrich iPSC from a variety of human source tissues.

  18. Reproductive isolation, reproductive mode, and sexual selection: empirical tests of the viviparity-driven conflict hypothesis.

    Science.gov (United States)

    Coleman, Seth W; Harlin-Cognato, April; Jones, Adam G

    2009-03-01

    A central goal in evolutionary biology is to elucidate general mechanisms and patterns of species divergence. The viviparity-driven conflict (VDC) hypothesis posits that intense mother-embryo conflict associated with viviparity drives rapid reproductive isolation among viviparous species, is intensified by multiple paternity, and reduces female reliance on precopulatory cues in mate choice. We tested these predictions using comparisons of oviparous and viviparous fishes. Consistent with the VDC hypothesis, we found that, relative to oviparous species, only closely related viviparous fishes are known to hybridize. Also in support of the VDC hypothesis, we found that (1) elaborate male sexual ornamentation may be more common in viviparous species with relatively low levels of maternal provisioning of embryos compared with those with high levels of provisioning and (2) the degree of multiple paternity is higher in viviparous species than in oviparous species. In contrast to a prediction of the VDC hypothesis, we found no relationship between the degree of multiple paternity and elaborate male sexual ornamentation, although statistical power was quite low for this test. Whereas overall our results strongly support the central tenet of the VDC hypothesis-that reproductive mode affects rates of evolution of reproductive isolation and the strength of sexual selection-they cannot rule out two alternative models we propose that may also explain the observed patterns.

  19. Nickel nanoparticle decorated graphene for highly selective isolation of polyhistidine-tagged proteins.

    Science.gov (United States)

    Liu, Jia-Wei; Yang, Ting; Ma, Lin-Yu; Chen, Xu-Wei; Wang, Jian-Hua

    2013-12-20

    Nickel nanoparticle decorated graphene (GP-Ni) is prepared by one-pot hydrothermal reduction of graphene oxide and nickel cations by hydrazine hydrate in the presence of poly(sodium-p-styrenesulfonate) (PSS). The GP-Ni hybrid is characterized by XRD, TEM, SEM, XPS, Raman and FT-IR spectra, demonstrating the formation of poly-dispersed nickel nanoparticles with an average size of 83 nm attached on the surface of graphene sheets. The GP-Ni hybrid exhibits ferromagnetic behavior with a magnetization saturation of 31.1 emu g(-1) at 10,000 Oersted (Oe). The GP-Ni also possesses favorable stability in aqueous medium and rapid magnetic response to an external magnetic field. These make it a novel magnetic adsorbent for the separation/isolation of His6-tagged recombinant proteins from a complex sample matrix (cell lysate). The targeted protein species is captured onto the surface of the GP-Ni hybrid via specific metal affinity force between polyhistidine groups and nickel nanoparticles. The SDS-PAGE assay indicates highly selective separation of His6-tagged Smt A from cell lysate. The GP-Ni hybrid displays favorable performance on the separation/isolation of His6-tagged recombinant proteins with respect to the commercial NTA-Ni(2+) column.

  20. A case of adult Pompe disease presenting with severe fatigue and selective involvement of type 1 muscle fibers.

    Science.gov (United States)

    van den Berg, Linda E M; de Vries, Juna M; Verdijk, Robert M; van der Ploeg, Ans T; Reuser, Arnold J J; van Doorn, Pieter A

    2011-03-01

    We present a case of adult Pompe disease (acid maltase deficiency) with an uncommon clinical presentation characterized by severe fatigue and myalgia prior to the onset of limb girdle weakness. Remarkably, the muscle biopsy demonstrated selective involvement of type 1 muscle fibers. The cause and clinical effects of fiber type specific involvement are currently unknown, but the phenomenon might contribute to the clinical heterogeneity in Pompe disease and the variable response to enzyme replacement therapy.

  1. Partial Sequencing of 16S rRNA Gene of Selected Staphylococcus aureus Isolates and its Antibiotic Resistance

    Directory of Open Access Journals (Sweden)

    Harsi Dewantari Kusumaningrum

    2016-08-01

    Full Text Available The choice of primer used in 16S rRNA sequencing for identification of Staphylococcus species found in food is important. This study aimed to characterize Staphylococcus aureus isolates by partial sequencing based on 16S rRNA gene employing primers 16sF, 63F or 1387R. The isolates were isolated from milk, egg dishes and chicken dishes and selected based on the presence of sea gene that responsible for formation of enterotoxin-A. Antibiotic susceptibility of the isolates towards six antibiotics was also tested. The use of 16sF resulted generally in higher identity percentage and query coverage compared to the sequencing by 63F or 1387R. BLAST results of all isolates, sequenced by 16sF, showed 99% homology to complete genome of four S. aureus strains, with different characteristics on enterotoxin production and antibiotic resistance. Considering that all isolates were carrying sea gene, indicated by the occurence of 120 bp amplicon after PCR amplification using primer SEA1/SEA2,  the isolates were most in agreeing to S. aureus subsp. aureus ST288. This study indicated that 4 out of 8 selected isolates were resistant towards streptomycin. The 16S rRNA gene sequencing using 16sF is useful for identification of S. aureus. However, additional analysis such as PCR employing specific gene target, should give a valuable supplementary information, when specific characteristic is expected.

  2. Environmental carbonate chemistry selects for phenotype of recently isolated strains of Emiliania huxleyi

    Science.gov (United States)

    Rickaby, Rosalind E. M.; Hermoso, Michaël; Lee, Renee B. Y.; Rae, Benjamin D.; Heureux, Ana M. C.; Balestreri, Cecilia; Chakravarti, Leela; Schroeder, Declan C.; Brownlee, Colin

    2016-05-01

    Coccolithophorid algae, particularly Emiliania huxleyi, are prolific biomineralisers that, under many conditions, dominate communities of marine eukaryotic plankton. Their ability to photosynthesise and form calcified scales (coccoliths) has placed them in a unique position in the global carbon cycle. Contrasting reports have been made with regards to the response of E. huxleyi to ocean acidification. Therefore, there is a pressing need to further determine the fate of this key organism in a rising CO2 world. In this paper, we investigate the phenotype of newly isolated, genetically diverse, strains of E. huxleyi from UK Ocean Acidification Research Programme (UKOA) cruises around the British Isles, the Arctic, and the Southern Ocean. We find a continuum of diversity amongst the physiological and photosynthetic parameters of different strains of E. huxleyi morphotype A under uniform, ambient conditions imposed in the laboratory. This physiology is best explained by adaptation to carbonate chemistry in the former habitat rather than being prescribed by genetic fingerprints such as the coccolithophore morphology motif (CMM). To a first order, the photosynthetic capacity of each strain is a function of both aqueous CO2 availability, and calcification rate, suggestive of a link between carbon concentrating ability and calcification. The calcification rate of each strain is related linearly to the natural environmental [CO32-] at the site of isolation, but a few exceptional strains display low calcification rates at the highest [CO32-] when calcification is limited by low CO2 availability and/or a lack of a carbon concentrating mechanism. We present O2-electrode measurements alongside coccolith oxygen isotopic composition and the uronic acid content (UAC) of the coccolith associated polysaccharide (CAP), that act as indirect tools to show the differing carbon concentrating ability of the strains. The environmental selection revealed amongst our recently isolated strain

  3. Anti-Depressant-Like Effect of Kaempferitrin Isolated from Justicia spicigera Schltdl (Acanthaceae in Two Behavior Models in Mice: Evidence for the Involvement of the Serotonergic System

    Directory of Open Access Journals (Sweden)

    Julia Cassani

    2014-12-01

    Full Text Available We evaluated the antidepressant-like effect of kaempferitrin (Km isolated from the plant Justicia spicigera (Asteraceae, which is used in traditional medicine for relieving emotional disorders, such as “la tristeza” (sadness or dysthymia and “el humor” (mood changes. The actions of Km were evaluated in a forced swimming test (FST and a suspension tail test (TST in mice. We explored the involvement of the serotonergic system and the hypothalamic-hypophysis-adrenal axis (HPA in the antidepressant-like effect of Km. To evaluate nonspecific effects of Km on general activity, the open field test (OFT was performed. Km at 5, 10, and 20 mg/kg induced an antidepressant-like effect. Sub-effective dose of Km (1 mg/kg produced a synergistic effect with imipramine (6.25 mg/kg and fluoxetine (10 mg/kg but not with desipramine (3.12 mg/kg. Pretreatment with p-chlorophenylalanine methyl ester (PCPA, a serotonin synthesis inhibitor, N-{2-(4-(2-methoxyphenyl-1-piperazinyl}-N-(2-pyridinylcyclohexecarboxamide (WAY-100635, a selective 5-HT1A receptor antagonist, and 8OH-DPAT, a selective 5-HT1A agonist, but not pindolol (10 mg/kg blocked the anti- immobility effect induced by Km. Taken together, these results indicate that the antidepressant-like effect of Km is related to the serotonergic system, principally 5-HT1A. This effect was not related to changes in locomotor activity.

  4. Anti-depressant-like effect of kaempferitrin isolated from Justicia spicigera Schltdl (Acanthaceae) in two behavior models in mice: evidence for the involvement of the serotonergic system.

    Science.gov (United States)

    Cassani, Julia; Dorantes-Barrón, Ana María; Novales, Lilian Mayagoitia; Real, Guadalupe Alva; Estrada-Reyes, Rosa

    2014-12-19

    We evaluated the antidepressant-like effect of kaempferitrin (Km) isolated from the plant Justicia spicigera (Asteraceae), which is used in traditional medicine for relieving emotional disorders, such as "la tristeza" (sadness or dysthymia) and "el humor" (mood changes). The actions of Km were evaluated in a forced swimming test (FST) and a suspension tail test (TST) in mice. We explored the involvement of the serotonergic system and the hypothalamic-hypophysis-adrenal axis (HPA) in the antidepressant-like effect of Km. To evaluate nonspecific effects of Km on general activity, the open field test (OFT) was performed. Km at 5, 10, and 20 mg/kg induced an antidepressant-like effect. Sub-effective dose of Km (1 mg/kg) produced a synergistic effect with imipramine (6.25 mg/kg) and fluoxetine (10 mg/kg) but not with desipramine (3.12 mg/kg). Pretreatment with p-chlorophenylalanine methyl ester (PCPA), a serotonin synthesis inhibitor, N-{2-(4-(2-methoxyphenyl)-1-piperazinyl}-N-(2-pyridinyl)cyclohexecarboxamide (WAY-100635), a selective 5-HT1A receptor antagonist, and 8OH-DPAT, a selective 5-HT1A agonist, but not pindolol (10 mg/kg) blocked the anti- immobility effect induced by Km. Taken together, these results indicate that the antidepressant-like effect of Km is related to the serotonergic system, principally 5-HT1A. This effect was not related to changes in locomotor activity.

  5. Unveiling of novel regio-selective fatty acid double bond hydratases from Lactobacillus acidophilus involved in the selective oxyfunctionalization of mono- and di-hydroxy fatty acids.

    Science.gov (United States)

    Kim, Kyoung-Rok; Oh, Hye-Jin; Park, Chul-Soon; Hong, Seung-Hye; Park, Ji-Young; Oh, Deok-Kun

    2015-11-01

    The aim of this study is the first time demonstration of cis-12 regio-selective linoleate double-bond hydratase. Hydroxylation of fatty acids, abundant feedstock in nature, is an emerging alternative route for many petroleum replaceable products thorough hydroxy fatty acids, carboxylic acids, and lactones. However, chemical route for selective hydroxylation is still quite challenging owing to low selectivity and many environmental concerns. Hydroxylation of fatty acids by hydroxy fatty acid forming enzymes is an important route for selective biocatalytic oxyfunctionalization of fatty acids. Therefore, novel fatty acid hydroxylation enzymes should be discovered. The two hydratase genes of Lactobacillus acidophilus were identified by genomic analysis, and the expressed two recombinant hydratases were identified as cis-9 and cis-12 double-bond selective linoleate hydratases by in vitro functional validation, including the identification of products and the determination of regio-selectivity, substrate specificity, and kinetic parameters. The two different linoleate hydratases were the involved enzymes in the 10,13-dihydroxyoctadecanoic acid biosynthesis. Linoleate 13-hydratase (LHT-13) selectively converted 10 mM linoleic acid to 13S-hydroxy-9(Z)-octadecenoic acid with high titer (8.1 mM) and yield (81%). Our study will expand knowledge for microbial fatty acid-hydroxylation enzymes and facilitate the designed production of the regio-selective hydroxy fatty acids for useful chemicals from polyunsaturated fatty acid feedstocks.

  6. [Simultaneous isolation of MRSA and Pseudomonas aeruginosa using a novel selective and differential PMAC agar].

    Science.gov (United States)

    Taguchi, F; Okuda, S; Uchino, U; Muraoka, H; Hasegawa, M; Kobayashi, I

    1996-09-01

    PMAC agar, a novel, selective and differential medium has been developed and was subjected for evaluation of its selective and differential capability of methicillin resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa from other bacteria such as Bacillus, Micrococcus, Gram-negative bacteria and drug resistant ones. Growth of MRSA and P. aeruginosa on PMAC agar was facilitated and their colonies were easily differentiated. Colonies of MRSA after 24 approximately 48 h incubation at 35 degrees C were small (2 to 4 mm in diameter), smooth and egg-yolk reaction positive. On the other hand, P. aeruginosa with pigment production (pyocianin, fluorescin or pyomelanin) formed large (2.5 to 7.0 mm in diameter), brownish black or brown colonies with a creamy edge. PMAC agar did not allow to grow unwanted bacteria tested except certain species formerly classified to Pseudomonas such as Burkholderia and Stenotrophomonas. However multi-drug resistant strains such as Enterobacter cloacae, Serratia marcescens and Acinetobacter calcoaceticus formed extremely small colonies. PMAC agar is recommended as a novel, useful medium for isolation, differentiation and presumptive identification of MRSA and P. aeruginosa from clinical and environmental sources.

  7. Magnetic Nanospheres Encapsulated by Mesoporous Copper Oxide Shell for Selective Isolation of Hemoglobin.

    Science.gov (United States)

    Guo, Zhi-Yong; Zhang, Yue; Zhang, Dan-Dan; Shu, Yang; Chen, Xu-Wei; Wang, Jian-Hua

    2016-11-02

    A novel strategy for the preparation of magnetic nanospheres encapsulated by mesoporous copper oxide shell, shortly termed as Fe3O4@mCuO, is reported via the calcination of Cu(NH3)4(NO3)2 into continuous mesoporous CuO shell onto the surface of Fe3O4 nanoparticles. The magnetic nanospheres are characterized to possess stable core-shell structure with a crystalline mesoporous CuO layer, exhibiting a CuO loading content of 25.2 ± 1.1% along with a favorable magnetic susceptibility. Fe3O4@mCuO nanospheres exhibit favorable selectivity on the adsorption of hemoglobin with a high adsorption capacity of up to 1162.5 mg g(-1). After adsorption, the high magnetic susceptibility allows convenient separation of the nanospheres by an external magnet. The retained hemoglobin could be readily recovered by using 0.5% (m/v) sodium dodecyl sulfate (SDS) as stripping reagent, providing a recovery of 78%. Circular dichroism spectra illustrate virtually no change in the conformation of hemoglobin after the process of adsorption/desorption. Fe3O4@mCuO nanospheres are further applied for the selective isolation of hemoglobin from human whole blood, achieving high-purity hemoglobin as demonstrated by SDS-PAGE (polyacrylamide gel electrophoresis) assays.

  8. Parallel signatures of selection in temporally isolated lineages of pink salmon.

    Science.gov (United States)

    Seeb, L W; Waples, R K; Limborg, M T; Warheit, K I; Pascal, C E; Seeb, J E

    2014-05-01

    Studying the effect of similar environments on diverse genetic backgrounds has long been a goal of evolutionary biologists with studies typically relying on experimental approaches. Pink salmon, a highly abundant and widely ranging salmonid, provide a naturally occurring opportunity to study the effects of similar environments on divergent genetic backgrounds due to a strict two-year semelparous life history. The species is composed of two reproductively isolated lineages with overlapping ranges that share the same spawning and rearing environments in alternate years. We used restriction-site-associated DNA (RAD) sequencing to discover and genotype approximately 8000 SNP loci in three population pairs of even- and odd-year pink salmon along a latitudinal gradient in North America. We found greater differentiation within the odd-year than within the even-year lineage and greater differentiation in the southern pair from Puget Sound than in the northern Alaskan population pairs. We identified 15 SNPs reflecting signatures of parallel selection using both a differentiation-based method (BAYESCAN) and an environmental correlation method (BAYENV). These SNPs represent genomic regions that may be particularly informative in understanding adaptive evolution in pink salmon and exploring how differing genetic backgrounds within a species respond to selection from the same natural environment.

  9. VARIABLE SELECTION BY PSEUDO WAVELETS IN HETEROSCEDASTIC REGRESSION MODELS INVOLVING TIME SERIES

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    A simple but efficient method has been proposed to select variables in heteroscedastic regression models. It is shown that the pseudo empirical wavelet coefficients corresponding to the significant explanatory variables in the regression models are clearly larger than those nonsignificant ones, on the basis of which a procedure is developed to select variables in regression models. The coefficients of the models are also estimated. All estimators are proved to be consistent.

  10. Ionic liquid-polyvinyl chloride ionomer for highly selective isolation of basic proteins.

    Science.gov (United States)

    Shu, Yang; Chen, Xu-Wei; Wang, Jian-Hua

    2010-04-15

    Hydrophilic ionic liquid-polyvinyl chloride (PVC) hybrids were prepared by immobilizing N-methylimidazole (N-mim) to PVC chains in toluene. The NmimCl-PVC hybrids were characterized by FT-IR, (1)H NMR, surface charge analysis and elemental analysis. The immobilization ratio, i.e., the percentage of chloride on PVC chain reacting with N-mim to form the hybrid, varies from 4.3% to 15.1% by increasing the N-mim/PVC molar ratio. The most distinct feature of the hybrid is its excellent selectivity for adsorbing basic proteins by effective suppression of the non-specific protein adsorption by pure PVC, and a higher immobilization ratio facilitates better selectivity. In Tris-HCl buffer, 100 microg mL(-1) of basic proteins, i.e., lysozyme (Lys), cytochrome c (cyt-c) and hemoglobin (Hb), were favorably adsorbed with efficiencies of 97%, 98% and 94% by the hybrid with an immobilization ratio of 15.1%, while the adsorption of acidic proteins, i.e., bovine albumin serum (BSA), transferring (Trf) and immunoglobulin G (IgG) were negligible. The retained Lys, cyt-c and Hb could be readily recovered by elution with phosphate buffer, carbonate buffer and SDS solution with efficiencies of 89%, 87% and 84%, respectively. Another feature of the hybrid is the significant improvement of the biocompatibility characterized by the maintenance of the activity of hemoglobin after adsorption and elution process. The practical usefulness of the hybrid was demonstrated by selective isolation of hemoglobin from human whole blood.

  11. ISOLATION OF ENDOTOXIN-SPECIFIC ANTIBODIES BY SELECTION OF AN SINGLE CHAIN PHAGE ANTIBODY LIBRARY

    Institute of Scientific and Technical Information of China (English)

    陈鸣; 俞丽丽; 张雪; 府伟灵

    2002-01-01

    Objective: To isolate murine anti endotoxin single chain phage antibody from a constructed library. Methods: Total RNA was firstly extracted from murine splenic cells and mRNA was reverse-transcribed into cDNA. Then the designed primers were used to amplify the variable region genes of the heavy and light chain (VH, VL) with polymerase chain reaction. The linker was used to assemble the VH and VL into ScFv, and the NotI and SfiI restriction enzymes were used to digest the ScFv in order to ligate into the pCANTAB5E phagemid vector that was already digested with the same restriction enzymes. The ligated vector was then introduced into competent E.coli TG1 cells to construct a single-chain phage antibody library. After rescued with M13KO7 helper phage, recombinant phages displaying ScFv fragments were harvested from the supernatant and selected with endotoxin. The enriched positive clones were reinfected into TG1 cells. Finally, 190 clones were randomly selected to detect the anti endotoxin antibody with indirect ELISA. Results: The titer of anti endotoxin in murine sera was 1:12,800. The concentration of total RNA was 12.38 μg/ml. 1.9×107 clones were obtained after transformed into TG1. 3×104 colonies were gotten after one round panning. Two positive colonies were confirmed with indirect ELISA among 190 randomly selected colonies. Conclusion: A 1.9×107 murine anti endotoxin single chain phage antibody library was successfully constructed. Two anti endotoxin antibodies were obtained from the library.

  12. Isolation gowns in health care settings: Laboratory studies, regulations and standards, and potential barriers of gown selection and use

    Science.gov (United States)

    Kilinc Balci, F. Selcen

    2016-01-01

    Although they play an important role in infection prevention and control, textile materials and personal protective equipment (PPE) used in health care settings are known to be one of the sources of cross-infection. Gowns are recommended to prevent transmission of infectious diseases in certain settings; however, laboratory and field studies have produced mixed results of their efficacy. PPE used in health care is regulated as either class I (low risk) or class II (intermediate risk) devices in the United States. Many organizations have published guidelines for the use of PPE, including isolation gowns, in health care settings. In addition, the Association for the Advancement of Medical Instrumentation published a guidance document on the selection of gowns and a classification standard on liquid barrier performance for both surgical and isolation gowns. However, there is currently no existing standard specific to isolation gowns that considers not only the barrier resistance but also a wide array of end user desired attributes. As a result, infection preventionists and purchasing agents face several difficulties in the selection process, and end users have limited or no information on the levels of protection provided by isolation gowns. Lack of knowledge about the performance of protective clothing used in health care became more apparent during the 2014 Ebola epidemic. This article reviews laboratory studies, regulations, guidelines and standards pertaining to isolation gowns, characterization problems, and other potential barriers of isolation gown selection and use. PMID:26391468

  13. Selection and RAPD analysis of Pseudomonas ssp. isolates able to improve biological viability of potato seed tubers

    Directory of Open Access Journals (Sweden)

    Poštić Dobrivoj

    2013-01-01

    Full Text Available Indoleacetic acid (IAA producing Pseudomonas isolates from the rhizosphere of maize (Q4 and Q20, alfalfa (Q1 and Q16 and wild red clover (B25 were selected for the investigation of their effect on the biological vitality of the potato seed tubers. The production of IAA ranged from 4.09 to 15.9 µ gmL- 1 after 24h of cultivation and 4.08 to 26.4 µ gmL-1 after 48h of cultivation. The molecular comparison by RAPD analysis also was done. RAPD patterns of selected Pseudomonas spp. isolates obtained by BC318, AF14, SPH 1 and AP 10 primers demonstrated the suitability of RAPD method in distinguishing a high variability among the four isolates (44 to 68%. The effect on the biological viability of potato (industrially important variety Pirol was observed during the seven weeks of sprouting at the temperature of 18-20°C. Potato tubers treated by the selected isolates formed slightly lower number of sprouts, but statistically higher mean length - up to 129.9% higher than the control. The mean sprouting capacity was 64.5% higher than a control. Obtained results suggested positive effects of selected IAA producing Pseudomonas isolates on the length of potato tubers and sprouting capacity as the parameters which define biological viability. [Projekat Ministarstva nauke Republike Srbije, br. III46007

  14. Selective memory retrieval of auditory what and auditory where involves the ventrolateral prefrontal cortex.

    Science.gov (United States)

    Kostopoulos, Penelope; Petrides, Michael

    2016-02-16

    There is evidence from the visual, verbal, and tactile memory domains that the midventrolateral prefrontal cortex plays a critical role in the top-down modulation of activity within posterior cortical areas for the selective retrieval of specific aspects of a memorized experience, a functional process often referred to as active controlled retrieval. In the present functional neuroimaging study, we explore the neural bases of active retrieval for auditory nonverbal information, about which almost nothing is known. Human participants were scanned with functional magnetic resonance imaging (fMRI) in a task in which they were presented with short melodies from different locations in a simulated virtual acoustic environment within the scanner and were then instructed to retrieve selectively either the particular melody presented or its location. There were significant activity increases specifically within the midventrolateral prefrontal region during the selective retrieval of nonverbal auditory information. During the selective retrieval of information from auditory memory, the right midventrolateral prefrontal region increased its interaction with the auditory temporal region and the inferior parietal lobule in the right hemisphere. These findings provide evidence that the midventrolateral prefrontal cortical region interacts with specific posterior cortical areas in the human cerebral cortex for the selective retrieval of object and location features of an auditory memory experience.

  15. Isolation of doublesex- and mab-3-related transcription factor 6 and its involvement in spermatogenesis in tilapia.

    Science.gov (United States)

    Zhang, Xianbo; Wang, Hai; Li, Minghui; Cheng, Yunying; Jiang, Dongneng; Sun, Lina; Tao, Wenjing; Zhou, Linyan; Wang, Zhijian; Wang, Deshou

    2014-12-01

    The dmrt6 gene has been isolated from tetrapods and recently from a coelacanth, Latimeria chalumnae. Its evolutionary history and exact function remain unclear. In the present study, dmrt6 was isolated from Perciformes (five cichlids and stickleback), Siluriformes (southern catfish), and Lepisosteiformes (spotted gar). Syntenic and phylogenetic analyses indicated that dmrt6 experienced gene transposition after the divergence of teleosts from other bony fish as gene loci surrounding dmrt6 were conserved among teleosts (but was completely different from gene loci surrounding dmrt6 in tetrapods and spotted gar), while these gene loci were conserved among nonteleost species. Real-time PCR and in situ hybridization revealed that dmrt6 was highly expressed in the XY gonads from 90 days after hatching (dah) onward and was observed exclusively in spermatocytes of the testes in tilapia. Dmrt6 knockout by CRISPR/Cas9 resulted in fewer spermatocytes, down-regulated Cyp11b2 in testes, and consequently produced a lower level of serum 11-ketotestosterone (11-KT) in Dmrt6-deficient XY fish compared with the XY control at 120 dah. From 150 to 180 dah, spermatogenesis gradually recovered, and cyp11b2 expression and serum 11-KT level were restored to the same levels as those of the XY control fish. In addition, a Dmrt6 mutation was observed in genomic DNA of sperm of G0 mutant fish and F1 fish. Taken together, our data suggest that dmrt6 also exists in bony fish. Its absence in most fish genomes was probably due to incomplete sequencing and/or secondary loss. The dmrt6 gene is highly expressed in spermatocytes and is involved in spermatogenesis in tilapia. © 2014 by the Society for the Study of Reproduction, Inc.

  16. Isolation

    DEFF Research Database (Denmark)

    Agerholm, Frank Juul

    2011-01-01

    Næringsstoffet har i dette nummer sat fokus på ”velvære i vinterkulden”, ”indendørsaktiviteter” og ”fedtafgift”. I klummen vises det, at disse tre fokusområder, der for en umiddelbar betragtning måske nok synes noget uensartede, falder sammen i ét tema: Isolation!......Næringsstoffet har i dette nummer sat fokus på ”velvære i vinterkulden”, ”indendørsaktiviteter” og ”fedtafgift”. I klummen vises det, at disse tre fokusområder, der for en umiddelbar betragtning måske nok synes noget uensartede, falder sammen i ét tema: Isolation!...

  17. Transient receptor potential A1 is involved in cold-induced contraction in the isolated rat colon smooth muscle.

    Science.gov (United States)

    Dong, Yang; Shi, Hai-Lian; Shi, Jian-Rong; Wu, Da-Zheng

    2010-08-25

    Transient receptor potential (TRP) A1, a member of TRP channel family, is activated by noxious cold. The aims of this study were to determine if TRPA1 contributed to cold-induced contractions in the isolated rat colon preparations and explore the potential mechanisms. The colon smooth muscle layers were surgically isolated from the male Wistar rats and changes in isotonic tension of longitudinal muscle under various treatments were recorded as colonic motilities. Cold stimuli were obtained by the reperfusion with Krebs-Henseleit solution at given temperature using Constant Flow Pump. The mRNA expressions of TRPA1, TRPV1 and TRPM8 in rat colon smooth muscle layer were examined by using reverse transcription-polymerase chain reaction (RT-PCR) techniques. The results showed that the contractions induced by cold stimuli (from 37 degrees C to 12 degrees C stepwise) were inversely proportional to the temperature with a maximum contraction at 17 degrees C in both proximal and distal colons (Pcolon smooth muscle layers. Cold-induced colonic contractions were specially inhibited by TRPA1 blocker, ruthenium red (30 μmol/L), in the proximal and distal colon (Pcolons (both PCA, 1 mmol/L). Extracellular calcium removal (EGTA, 1 mmol/L), PLC blocker (U73122, 10 μmol/L) and IP(3) receptor blocker (2-aminoethoxydiphenyl borate, 2-APB, 30 μmol/L) all decreased the contractions evoked by the cooling at 17 degrees C in the proximal and distal colon (PCa(2+) channels blocker nifedipine (1 μmol/L) and neurotoxin tetrodotoxin (TTX, 2 μmol/L) decreased the contractile response in the distal colon (Pcolon. In conclusion, TRPA1 contributes to cold-induced contractions of the rat colon smooth muscle, and the mechanism of TRPA1 activation involves PLC/IP(3)/Ca(2+) pathway. L-type Ca(2+) channel and neurogenic mechanism other than muscarinic receptor might be partially involved in cold-induced contraction of the distal colon, which probably resulted in higher contraction of distal colon

  18. Replication of clinical hepatitis B virus isolate and its application for selecting antiviral agents for chronic hepatitis B patients

    Institute of Scientific and Technical Information of China (English)

    Yin-ping Lu; Tao Guo; Bao-Ju Wang; Ji-Hua Dong; Jian-Fang Zhu; Zhao Liu; Meng-Ji Lu; Dong-Liang Yang

    2008-01-01

    AIM: To establish a cell model harboring replicative clinical hepatitis B virus (HBV) isolates and evaluate its application in individualized selection of anti-HBV agents for chronic hepatitis B (CHB) Patients.METHODS: The full-length HBV genomic DNA from 8 CHB patients was amplified by polymerase chain reaction (PCR). All the patients were treated with lamivudine for at least seven months and finally became resistant to lamivudine. The amplified HBV DNA fragments were inserted into pHY106 vectors by Sap digestion. The recombinant plasmids containing 1.1 copies of HBV genome were transiently transfected into Huh7 cell line, and the levels of HBsAg, HBeAg and intercellular HBV replicative intermediates were determined by ELISA and Southern blot analysis, respectively, with or without lamivudine and adefovir treatment. The antiviral treatment with adefovir was administered to the patients and analyzed in parallel.RESULTS: A total of 25 independent HBV isolates were obtained from the sera of 8 patients, each patient had at least two isolates. One isolate from each individual was selected and subcloned into pHY106 vector, including 5 isolates with YVDD mutation and 3 isolates with YIDD mutation. All recombinant plasmids harboring HBV isolates were transfected into Huh7 cells. The results indicated that HBV genome carried in HBV replicons of clinical HBV isolates could effectively replicate and express in Huh7 cells. Adefovir, but not lamivudine, inhibited HBV replication both in vitro and in vivo, and in vitro inhibition was dose-dependent.CONCLUSION: The novel method described herein enables individualized selection of anti-HBV agents in clinic and is useful in future studies of antiviral therapy for CHB.

  19. Selection and characterization of Argentine isolates of Trichoderma harzianum for effective biocontrol of Septoria leaf blotch of wheat.

    Science.gov (United States)

    Stocco, Marina C; Mónaco, Cecilia I; Abramoff, Cecilia; Lampugnani, Gladys; Salerno, Graciela; Kripelz, Natalia; Cordo, Cristina A; Consolo, Verónica F

    2016-03-01

    Species of the genus Trichoderma are economically important as biocontrol agents, serving as a potential alternative to chemical control. The applicability of Trichoderma isolates to different ecozones will depend on the behavior of the strains selected from each zone. The present study was undertaken to isolate biocontrol populations of Trichoderma spp. from the Argentine wheat regions and to select and characterize the best strains of Trichoderma harzianum by means of molecular techniques. A total of 84 out of the 240 strains of Trichoderma were able to reduce the disease severity of the leaf blotch of wheat. Thirty-seven strains were selected for the reduction equal to or greater than 50% of the severity, compared with the control. The percentage values of reduction of the pycnidial coverage ranged between 45 and 80%. The same last strains were confirmed as T. harzianum by polymerase chain reaction amplification of internal transcribed spacers, followed by sequencing. Inter-simple sequence repeat was used to examine the genetic variability among isolates. This resulted in a total of 132 bands. Further numerical analysis revealed 19 haplotypes, grouped in three clusters (I, II, III). Shared strains, with different geographical origins and isolated in different years, were observed within each cluster. The origin of the isolates and the genetic group were partially related. All isolates from Paraná were in cluster I, all isolates from Lobería were in cluster II, and all isolates from Pergamino and Santa Fe were in cluster III. Our results suggest that the 37 native strains of T. harzianum are important in biocontrol programs and could be advantageous for the preparation of biopesticides adapted to the agroecological conditions of wheat culture.

  20. Selection of appropriate isolation method based on morphology of blastocyst for efficient derivation of buffalo embryonic stem cells.

    Science.gov (United States)

    Kumar, R; Ahlawat, S P S; Sharma, M; Verma, O P; Sai Kumar, G; Taru Sharma, G

    2014-03-01

    The efficiency of embryonic stem cell (ESC) derivation from all species except for rodents and primates is very low. There are however, multiple interests in obtaining pluripotent cells from these animals with main expectations in the fields of transgenesis, cloning, regenerative medicine and tissue engineering. Researches are being carried out in laboratories throughout the world to increase the efficiency of ESC isolation for their downstream applications. Thus, the present study was undertaken to study the effect of different isolation methods based on the morphology of blastocyst for efficient derivation of buffalo ESCs. Embryos were produced in vitro through the procedures of maturation, fertilization and culture. Hatched blastocysts or isolated inner cell masses (ICMs) were seeded on mitomycin-C inactivated buffalo fetal fibroblast monolayer for the development of ESC colonies. The ESCs were analyzed for alkaline phosphatase activity, expression of pluripotency markers and karyotypic stability. Primary ESC colonies were obtained after 2-5 days of seeding hatched blastocysts or isolated ICMs on mitomycin-C inactivated feeder layer. Mechanically isolated ICMs attached and formed primary cell colonies more efficiently than ICMs isolated enzymatically. For derivation of ESCs from poorly defined ICMs intact hatched blastocyst culture was the most successful method. Results of this study implied that although ESCs can be obtained using all three methods used in this study, efficiency varies depending upon the morphology of blastocyst and isolation method used. So, appropriate isolation method must be selected depending on the quality of blastocyst for efficient derivation of ESCs.

  1. Role of melanin in release of extracellular enzymes and selection of aggressive isolates of Bipolaris sorokiniana in barley.

    Science.gov (United States)

    Chand, Ramesh; Kumar, Manoj; Kushwaha, Chanda; Shah, Kavita; Joshi, Arun K

    2014-08-01

    Eighteen barley isolates of Bipolaris sorokiniana belonging to wild and clonal type of black, mixed and white subpopulations were quantitatively assayed for their melanin content and aggressiveness with respect to production of some of the extracellular enzymes such as cellulase, pectinase, amylase and protease. Cellulase and pectinase constituted major portion of the enzymes recovered from the black, mixed and white isolates. Enzyme production and aggressiveness were relatively higher in melanin devoid or low melanin isolates. The melanin deficient isolates were also differentiated from black and mixed isolates on the basis of variation in internal transcribed spacer region of the ribosomal DNA. Higher enzyme productions positively correlated with area under disease progress curve (AUDPC) and lesion development. Melanin content was negatively correlated with extracellular enzymes and aggressiveness of the isolates. Based on melanin content, lesion size, AUDPC and extracellular enzymes, the isolates were grouped in two major clusters (I and II) with further division of cluster II into two sub-clusters (II-A and II-B). The results appears to indicate a possible role of melanin in release of extracellular enzymes and hence in evolution and selection of aggressive isolates of B. sorokiniana in barley.

  2. Selection and Evaluation of Attorneys in Divorce Cases involving Minor Children.

    Science.gov (United States)

    Hillary, Marcia Amanda; Johnson, Joel T.

    1985-01-01

    Examines reasons for hiring a divorce attorney, the attorney selection process, and client evaluations of their attorneys. Sixty-six men and women, who were parents of at least one minor child, indicated that they learned about their divorce attorney through a personal recommendation, and hired the first and only lawyer whom they consulted.…

  3. Age and gender differences in mate selection criteria for various involvement levels

    NARCIS (Netherlands)

    Buunk, Abraham (Bram); Dijkstra, Pieternel; Fetchenhauer, D.; Kenrick, D.T.

    2002-01-01

    The present study investigated mate preferences for five different levels of relationship involvement-marriage, serious relationship, falling in love, casual sex, and sexual fantasies-among individuals of 20, 30, 40, 50, and 60 years of age. Consistent with an evolutionary perspective, men preferred

  4. Age and gender differences in mate selection criteria for various involvement levels

    NARCIS (Netherlands)

    Buunk, Abraham (Bram); Dijkstra, Pieternel; Fetchenhauer, D.; Kenrick, D.T.

    The present study investigated mate preferences for five different levels of relationship involvement-marriage, serious relationship, falling in love, casual sex, and sexual fantasies-among individuals of 20, 30, 40, 50, and 60 years of age. Consistent with an evolutionary perspective, men preferred

  5. Parent Involvement in Early Childhood Special Education: Selected Activities for Preschool Teachers.

    Science.gov (United States)

    Carlson, Betty Clark

    The manual is designed to provide special education preschool teachers with a guide for parent involvement activities. The guide presents 32 activities divided into three topical areas: (1) communicating with parents (orientation packet, newsletter, school-home notebook, bulletin board for parents); (2) resource activities (field trips, home…

  6. Calcium paradox induces apoptosis in the isolated perfused Rana ridibunda heart: involvement of p38-MAPK and calpain.

    Science.gov (United States)

    Aggeli, Ioanna-Katerina; Zacharias, Triantafyllos; Papapavlou, Georgia; Gaitanaki, Catherine; Beis, Isidoros

    2013-12-01

    "Calcium paradox" as a term describes the deleterious effects conferred to a heart perfused with a calcium-free solution followed by repletion, including loss of mechanical activity and sarcomere disruption. Given that the signaling mechanisms triggered by calcium paradox remain elusive, in the present study, we tried to investigate them in the isolated perfused heart from Rana ridibunda. Calcium paradox was found to markedly activate members of the MAPKs (p43-ERK, JNKs, p38-MAPK). In addition to lactate dehydrogenase (LDH) release in the perfusate (indicative of necrosis), we also confirmed the occurrence of apoptosis by using the TUNEL assay and identifying poly(ADP-ribose) polymerase (PARP) fragmentation and upregulated Bax expression. Furthermore, using MDL28170 (a selective calpain inhibitor), a role for this protease was revealed. In addition, various divalent cations were shown to exert a protective effect against the calcium paradox. Interestingly, SB203580, a p38-MAPK inhibitor, alleviated calcium-paradox-conferred apoptosis. This result indicates that p38-MAPK plays a pro-apoptotic role, contributing to the resulting myocardial dysfunction and cell death. To our knowledge, this is the first time that the calcium paradox has been shown to induce apoptosis in amphibians, with p38-MAPK and calpain playing significant roles.

  7. Postzygotic isolation involves strong mitochondrial and sex-specific effects in Tigriopus californicus, a species lacking heteromorphic sex chromosomes.

    Science.gov (United States)

    Foley, B R; Rose, C G; Rundle, D E; Leong, W; Edmands, S

    2013-11-01

    Detailed studies of the genetics of speciation have focused on a few model systems, particularly Drosophila. The copepod Tigriopus californicus offers an alternative that differs from standard animal models in that it lacks heteromorphic chromosomes (instead, sex determination is polygenic) and has reduced opportunities for sexual conflict, because females mate only once. Quantitative trait loci (QTL) mapping was conducted on reciprocal F2 hybrids between two strongly differentiated populations, using a saturated linkage map spanning all 12 autosomes and the mitochondrion. By comparing sexes, a possible sex ratio distorter was found but no sex chromosomes. Although studies of standard models often find an excess of hybrid male sterility factors, we found no QTL for sterility and multiple QTL for hybrid viability (indicated by non-Mendelian adult ratios) and other characters. Viability problems were found to be stronger in males, but the usual explanations for weaker hybrid males (sex chromosomes, sensitivity of spermatogenesis, sexual selection) cannot fully account for these male viability problems. Instead, higher metabolic rates may amplify deleterious effects in males. Although many studies of standard speciation models find the strongest genetic incompatibilities to be nuclear-nuclear (specifically X chromosome-autosome), we found the strongest deleterious interaction in this system was mito-nuclear. Consistent with the snowball theory of incompatibility accumulation, we found that trigenic interactions in this highly divergent cross were substantially more frequent (>6×) than digenic interactions. This alternative system thus allows important comparisons to studies of the genetics of reproductive isolation in more standard model systems.

  8. Isolation of arginine kinase from Apis cerana cerana and its possible involvement in response to adverse stress.

    Science.gov (United States)

    Chen, Xiaobo; Yao, Pengbo; Chu, Xiaoqian; Hao, Lili; Guo, Xingqi; Xu, Baohua

    2015-01-01

    Arginine kinases (AK) in invertebrates play the same role as creatine kinases in vertebrates. Both proteins are important for energy metabolism, and previous studies on AK focused on this attribute. In this study, the arginine kinase gene was isolated from Apis cerana cerana and was named AccAK. A 5'-flanking region was also cloned and shown to contain abundant putative binding sites for transcription factors related to development and response to adverse stress. We imitated several abiotic and biotic stresses suffered by A. cerana cerana during their life, including heavy metals, pesticides, herbicides, heat, cold, oxidants, antioxidants, ecdysone, and Ascosphaera apis and then studied the expression patterns of AccAK after these treatments. AccAK was upregulated under all conditions, and, in some conditions, this response was very pronounced. Western blot and AccAK enzyme activity assays confirmed the results. In addition, a disc diffusion assay showed that overexpression of AccAK reduced the resistance of Escherichia coli cells to multiple adverse stresses. Taken together, our results indicated that AccAK may be involved of great significance in response to adverse abiotic and biotic stresses.

  9. Selective isolation and differentiation of a stromal population of human embryonic stem cells with osteogenic potential

    DEFF Research Database (Denmark)

    Harkness, Linda M; Mahmood, Amer; Ditzel, Nicholas

    2011-01-01

    The derivation of osteogenic cells from human embryonic stem cells (hESC) has been hampered by the absence of easy and reproducible protocols. hESC grown in feeder-free conditions, often show a sub population of fibroblast-like, stromal cells growing between the colonies. Thus, we examined...... the possibility that these cells represent a population of stromal (mesenchymal) stem cells (hESC-stromal). Two in house derived hES cell lines (Odense3 and KMEB3) as well as an externally derived cell line (Hues8) were transitioned to feeder-free conditions. A sub population of fibroblast-like cells established...... between the hESC colonies were isolated by selective adherence to hyaluronic acid-coated plates (100μg/ml) and were characterized using a combination of FACS analysis and staining. The cells were CD44(+), CD29(+), CD73(+), CD166(+), CD146(+), and CD105(+); and, Oct4(-), CD34(-), CD45(-) and CXCR4(-). When...

  10. The Use of Variable Q1 Isolation Windows Improves Selectivity in LC-SWATH-MS Acquisition.

    Science.gov (United States)

    Zhang, Ying; Bilbao, Aivett; Bruderer, Tobias; Luban, Jeremy; Strambio-De-Castillia, Caterina; Lisacek, Frédérique; Hopfgartner, Gérard; Varesio, Emmanuel

    2015-10-02

    As tryptic peptides and metabolites are not equally distributed along the mass range, the probability of cross fragment ion interference is higher in certain windows when fixed Q1 SWATH windows are applied. We evaluated the benefits of utilizing variable Q1 SWATH windows with regards to selectivity improvement. Variable windows based on equalizing the distribution of either the precursor ion population (PIP) or the total ion current (TIC) within each window were generated by an in-house software, swathTUNER. These two variable Q1 SWATH window strategies outperformed, with respect to quantification and identification, the basic approach using a fixed window width (FIX) for proteomic profiling of human monocyte-derived dendritic cells (MDDCs). Thus, 13.8 and 8.4% additional peptide precursors, which resulted in 13.1 and 10.0% more proteins, were confidently identified by SWATH using the strategy PIP and TIC, respectively, in the MDDC proteomic sample. On the basis of the spectral library purity score, some improvement warranted by variable Q1 windows was also observed, albeit to a lesser extent, in the metabolomic profiling of human urine. We show that the novel concept of "scheduled SWATH" proposed here, which incorporates (i) variable isolation windows and (ii) precursor retention time segmentation further improves both peptide and metabolite identifications.

  11. Antibacterial Action of Jineol Isolated from Scolopendra subspinipes mutilans against Selected Foodborne Pathogens

    Science.gov (United States)

    Bajpai, Vivek K.; Shukla, Shruti; Paek, Woon K.; Lim, Jeongheui; Kumar, Pradeep; Na, MinKyun

    2017-01-01

    This study was undertaken to assess the antibacterial potential of 3,8-dihydroxyquinoline (jineol) isolated from Scolopendra subspinipes mutilans against selected foodborne pathogens Escherichia coli O157:H7 and Staphylococcus aureus KCTC-1621. Jineol at the tested concentration (50 μL; corresponding to 250 μg/disk) exhibited significant antibacterial effects as a diameter of inhibition zones (11.6–13.6 mm), along with minimum inhibitory concentration (MIC) and minimum bactericidal concentration values found in the range of (62.5–125 μg/mL) and (125–250 μg/mL), respectively. Jineol also exhibited significant antibacterial effects as confirmed by the reduction in bacterial cell viabilities, increasing release of potassium (K+) ions (650 and 700 mmole/L) and 260 nm materials (optical density: 2.98–3.12) against both the tested pathogens, E. coli O157:H7 and S. aureus KCTC-1621, respectively. Moreover, changes in the cell wall morphology of E. coli O157:H7 and S. aureus KCTC-1621 cells treated with jineol at MIC further confirmed its inhibitory potential against the tested pathogens, suggesting its role as an effective antimicrobial to control foodborne pathogens.

  12. Selected, annotated bibliography of studies relevant to the isolation of nuclear wastes. [705 references

    Energy Technology Data Exchange (ETDEWEB)

    Hyder, L.K.; Fore, C.S.; Vaughan, N.D.; Faust, R.A.

    1980-09-01

    This annotated bibliography of 705 references represents the first in a series to be published by the Ecological Sciences Information Center containing scientific, technical, economic, and regulatory information relevant to nuclear waste isolation. Most references discuss deep geologic disposal, with fewer studies of deep seabed disposal; space disposal is also included. The publication covers both domestic and foreign literature for the period 1954 to 1980. Major chapters selected are Chemical and Physical Aspects; Container Design and Performance; Disposal Site; Envirnmental Transport; General Studies and Reviews; Geology, Hydrology and Site Resources; Regulatory and Economic Aspects; Repository Design and Engineering; Transportation Technology; Waste Production; and Waste Treatment. Specialized data fields have been incorporated to improve the ease and accuracy of locating pertinent references. Specific radionuclides for which data are presented are listed in the Measured Radionuclides field, and specific parameters which affect the migration of these radionuclides are presented in the Measured Parameters field. The references within each chapter are arranged alphabetically by leading author, corporate affiliation, or title of the document. When the author is not given, the corporate affiliation appears first. If these two levels of authorship are not given, the title of the document is used as the identifying level. Indexes are provided for author(s), keywords, subject category, title, geographic location, measured parameters, measured radionuclides, and publication description.

  13. Using selective chromogenic plates to optimize isolation of group B Streptococcus in pregnant women

    Directory of Open Access Journals (Sweden)

    Romano Mattei

    2014-03-01

    Full Text Available Group B Streptococcus (GBS remains the leading cause of severe bacterial infections (sepsis, meningitis, pneumonia in neonates. We compared the detection of GBS from recto-vaginal swabs on blood agar and two chromogenic media and evaluated their antibiotic susceptibility. A total of 1351 swabs were taken from pregnant women at 35-37 weeks of gestation. Following enrichment in Todd Hewitt broth + nalidixic acid and colistin, the samples were plated on Columbia CNA agar (CNA, chromID Strepto B agar (STRB and Granada Agar (GRAN, respectively. GBS were found in 22.4% of recto-vaginal swabs from pregnant women. Sensitivity, specificity, positive and negative predictive values of GBS detection were 88%, 88%, 81% and 96% for CNA, 99%, 97%, 90% and 99% for STRB and 94%, 99%, 98% e 99% for GRAN; Cohen’s k index concordances for CNA, STREB and GRAN were 0.68, 0.92 and 0.96, respectively. All isolates were susceptible to penicillin, whereas resistances of erythromycin and clindamycin were 40% and 42%, respectively. To conclude, selective broth enrichment combined with chromogenic plates is recommended for GBS screening in pregnant women.

  14. Phylodynamic Analysis of Clinical and Environmental Vibrio cholerae Isolates from Haiti Reveals Diversification Driven by Positive Selection

    Science.gov (United States)

    Azarian, Taj; Ali, Afsar; Johnson, Judith A.; Mohr, David; Prosperi, Mattia; Veras, Nazle M.; Jubair, Mohammed; Strickland, Samantha L.; Rashid, Mohammad H.; Alam, Meer T.; Weppelmann, Thomas A.; Katz, Lee S.; Tarr, Cheryl L.; Colwell, Rita R.

    2014-01-01

    ABSTRACT Phylodynamic analysis of genome-wide single-nucleotide polymorphism (SNP) data is a powerful tool to investigate underlying evolutionary processes of bacterial epidemics. The method was applied to investigate a collection of 65 clinical and environmental isolates of Vibrio cholerae from Haiti collected between 2010 and 2012. Characterization of isolates recovered from environmental samples identified a total of four toxigenic V. cholerae O1 isolates, four non-O1/O139 isolates, and a novel nontoxigenic V. cholerae O1 isolate with the classical tcpA gene. Phylogenies of strains were inferred from genome-wide SNPs using coalescent-based demographic models within a Bayesian framework. A close phylogenetic relationship between clinical and environmental toxigenic V. cholerae O1 strains was observed. As cholera spread throughout Haiti between October 2010 and August 2012, the population size initially increased and then fluctuated over time. Selection analysis along internal branches of the phylogeny showed a steady accumulation of synonymous substitutions and a progressive increase of nonsynonymous substitutions over time, suggesting diversification likely was driven by positive selection. Short-term accumulation of nonsynonymous substitutions driven by selection may have significant implications for virulence, transmission dynamics, and even vaccine efficacy. PMID:25538191

  15. Isolation, Identification, and Antimicrobial Susceptibility of Bacteria Associated with Waterpipe Contaminants in Selected Area of Saudi Arabia.

    Science.gov (United States)

    Alaidarous, Mohammed; Alanazi, Meshal; Abdel-Hadi, Ahmed

    2017-01-01

    This study highlights the level of microbial contamination of waterpipe components in selected area of Saudi Arabia and the resistance of selected bacteria to different antibiotics was determined. A series of biochemical tests, microscopic examination, and screening on Vitek 2 compact (bioMérieux Inc., USA) system were done to characterize the bacterial isolates. Out of 132 samples investigated, 7 mouthpiece samples and 48 water bowl samples showed positivity on culture. The percentage of contamination rate was higher in water bowl (69.69%) than in mouthpieces (10.6%) for all selected areas. A total of 55 bacterial isolates were identified which included Gram-negative (28) and Gram-positive (27) bacteria. Antimicrobial susceptibility data showed more resistance to bacteria isolated from water bowl than bacteria isolated from mouthpiece. In addition, one isolate which was confirmed as methicillin-resistant Staphylococcus aureus and Klebsiella pneumoniae was resistant to antibiotics which are commonly used to treat pneumonia. Water bowl of waterpipe instrument is significantly contaminated with different bacterial pathogens including multidrug-resistant and pneumonia causing bacteria, which are a real health concern among waterpipe smokers. The presented data could assist public health professionals to raise the concerns regarding cleaning practices of waterpipe components and highlights the risk posed among the waterpipe smokers.

  16. Selective importance of the rat anterior thalamic nuclei for configural learning involving distal spatial cues.

    Science.gov (United States)

    Dumont, Julie R; Amin, Eman; Aggleton, John P

    2014-01-01

    To test potential parallels between hippocampal and anterior thalamic function, rats with anterior thalamic lesions were trained on a series of biconditional learning tasks. The anterior thalamic lesions did not disrupt learning two biconditional associations in operant chambers where a specific auditory stimulus (tone or click) had a differential outcome depending on whether it was paired with a particular visual context (spot or checkered wall-paper) or a particular thermal context (warm or cool). Likewise, rats with anterior thalamic lesions successfully learnt a biconditional task when they were reinforced for digging in one of two distinct cups (containing either beads or shredded paper), depending on the particular appearance of the local context on which the cup was placed (one of two textured floors). In contrast, the same rats were severely impaired at learning the biconditional rule to select a specific cup when in a particular location within the test room. Place learning was then tested with a series of go/no-go discriminations. Rats with anterior thalamic nuclei lesions could learn to discriminate between two locations when they were approached from a constant direction. They could not, however, use this acquired location information to solve a subsequent spatial biconditional task where those same places dictated the correct choice of digging cup. Anterior thalamic lesions produced a selective, but severe, biconditional learning deficit when the task incorporated distal spatial cues. This deficit mirrors that seen in rats with hippocampal lesions, so extending potential interdependencies between the two sites.

  17. Selective involvement of BH3-only proteins and differential targets of Noxa in diverse apoptotic pathways.

    Science.gov (United States)

    Zhang, L; Lopez, H; George, N M; Liu, X; Pang, X; Luo, X

    2011-05-01

    The BH3-only proteins of the Bcl-2 family are known to mediate mitochondrial dysfunction during apoptosis. However, the identity of the critical BH3-only proteins and the mechanism of their action following treatment by diverse apoptotic stimuli remain to be fully resolved. We therefore used RNAi to screen the entire Bcl-2 family for their involvement in three major apoptotic pathways in HeLa cells. We found that Bcl-xL and Mcl-1 are major inhibitors of apoptosis induced by TNF-related apoptosis-inducing ligand (TRAIL), endoplasmic reticulum (ER) stress, and proteasome inhibition. Among the 10 BH3-only proteins, Bid and Noxa were found to be critically involved in TRAIL-induced apoptosis, in which Noxa participates by constitutively binding to Mcl-1. Bim and Noxa were found to be necessary for ER stress-induced apoptosis, in which Noxa assisted Bim function by sequestering Mcl-1 and binding to Bcl-xL. As a critical BH3-only protein, Noxa was strongly upregulated and became associated with both Mcl-1 and Bcl-xL during apoptosis induced by proteasome inhibition. In addition, we found that Noxa became 'Mcl-1 free' following treatment by ER stress and proteasome inhibition, but not after TRAIL treatment. These results defined the critical Bcl-2 network during apoptosis and suggested that Noxa participated in triggering mitochondrial dysfunction in multiple apoptotic pathways through distinct mechanisms.

  18. Selected fault testing of electronic isolation devices used in nuclear power plant operation

    Energy Technology Data Exchange (ETDEWEB)

    Villaran, M.; Hillman, K.; Taylor, J.; Lara, J.; Wilhelm, W. [Brookhaven National Lab., Upton, NY (United States)

    1994-05-01

    Electronic isolation devices are used in nuclear power plants to provide electrical separation between safety and non-safety circuits and systems. Major fault testing in an earlier program indicated that some energy may pass through an isolation device when a fault at the maximum credible potential is applied in the transverse mode to its output terminals. During subsequent field qualification testing of isolators, concerns were raised that the worst case fault, that is, the maximum credible fault (MCF), may not occur with a fault at the maximum credible potential, but rather at some lower potential. The present test program investigates whether problems can arise when fault levels up to the MCF potential are applied to the output terminals of an isolator. The fault energy passed through an isolated device during a fault was measured to determine whether the levels are great enough to potentially damage or degrade performance of equipment on the input (Class 1E) side of the isolator.

  19. Involvement of AMPA/kainate and GABAA receptors in topiramate neuroprotective effects against methylphenidate abuse sequels involving oxidative stress and inflammation in rat isolated hippocampus.

    Science.gov (United States)

    Motaghinejad, Majid; Motevalian, Manijeh

    2016-08-01

    Abuses of methylphenidate (MPH) as psychostimulant cause neural damage of brain cells. Neuroprotective properties of topiramate (TPM) have been indicated in several studies but its exact mechanism of action remains unclear. The current study evaluates protective role of various doses of TPM and its mechanism of action in MPH induced oxidative stress and inflammation. The neuroprotective effects of various doses of TPM against MPH induced oxidative stress and inflammation were evaluated and then the action of TPM was studied in presence of domoic acid (DOM), as AMPA/kainate receptor agonist and bicuculline (BIC) as GABAA receptor antagonist, in isolated rat hippocampus. Open Field Test (OFT) was used to investigate motor activity changes. Oxidative, antioxidant and inflammatory factors were measured in isolated hippocampus. TPM (70 and 100mg/kg) decreased MPH induced motor activity disturbances and inhibit MPH induced oxidative stress and inflammation. On the other hand pretreatment of animals with DOM or BIC, inhibit this effect of TPM and potentiate MPH induced motor activity disturbances and increased lipid peroxidation, mitochondrial oxidized form of glutathione (GSSG) level, interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in isolated hippocampal cells and decreased reduced form of glutathione (GSH) level, superoxide dismutase, glutathione peroxidase and glutathione reductase activity. It seems that TPM can protect cells of hippocampus from oxidative stress and neuroinflammation and it could be partly by activation of GABAA receptor and inhibition of AMPA/kainite receptor.

  20. Application of the ELECTRE I method for selection problems involving free software development projects

    Directory of Open Access Journals (Sweden)

    Jéfferson Jesus de Araujo

    2016-06-01

    Full Text Available The research is aimed at practical application of the ELECTRE I method for decision-making support in the selection of projects by a free software development company in the city of Juazeiro - BA. The methodology used in the company, the projects to be evaluated, the criteria chosen by the decision maker and their respective weights were used to model the problem, by building decision matrix and conducting ELECTRE I. The results proved effective, satisfying the proposal made in the research. Finally, the conclusions are presented, provoking a discussion of the current method and the proposed method, suggesting the application of the ELECTRE I method and other MCDA methods for other decision-making problems at the company.

  1. Bilingualism trains specific brain circuits involved in flexible rule selection and application.

    Science.gov (United States)

    Stocco, Andrea; Prat, Chantel S

    2014-10-01

    Bilingual individuals have been shown to outperform monolinguals on a variety of tasks that measure non-linguistic executive functioning, suggesting that some facets of the bilingual experience give rise to generalized improvements in cognitive performance. The current study investigated the hypothesis that such advantage in executive functioning arises from the need to flexibly select and apply rules when speaking multiple languages. Such flexible behavior may strengthen the functioning of the fronto-striatal loops that direct signals to the prefrontal cortex. To test this hypothesis, we compared behavioral and brain data from proficient bilinguals and monolinguals who performed a Rapid Instructed Task Learning paradigm, which requires behaving according to ever-changing rules. Consistent with our hypothesis, bilinguals were faster than monolinguals when executing novel rules, and this improvement was associated with greater modulation of activity in the basal ganglia. The implications of these findings for language and executive function research are discussed herein.

  2. Identification and Characterization of Genes Involved in Leishmania Pathogenesis: The Potential for Drug Target Selection

    Directory of Open Access Journals (Sweden)

    Robert Duncan

    2011-01-01

    Full Text Available Identifying and characterizing Leishmania donovani genes and the proteins they encode for their role in pathogenesis can reveal the value of this approach for finding new drug targets. Effective drug targets are likely to be proteins differentially expressed or required in the amastigote life cycle stage found in the patient. Several examples and their potential for chemotherapeutic disruption are presented. A pathway nearly ubiquitous in living cells targeted by anticancer drugs, the ubiquitin system, is examined. New findings in ubiquitin and ubiquitin-like modifiers in Leishmania show how disruption of those pathways could point to additional drug targets. The programmed cell death pathway, now recognized among protozoan parasites, is reviewed for some of its components and evidence that suggests they could be targeted for antiparasitic drug therapy. Finally, the endoplasmic reticulum quality control system is involved in secretion of many virulence factors. How disruptions in this pathway reduce virulence as evidence for potential drug targets is presented.

  3. Positive selection on NIN, a gene involved in neurogenesis, and primate brain evolution.

    Science.gov (United States)

    Montgomery, S H; Mundy, N I

    2012-11-01

    A long-held dogma in comparative neurobiology has been that the number of neurons under a given area of cortical surface is constant. As such, the attention of those seeking to understand the genetic basis of brain evolution has focused on genes with functions in the lateral expansion of the developing cerebral cortex. However, new data suggest that cortical cytoarchitecture is not constant across primates, raising the possibility that changes in radial cortical development played a role in primate brain evolution. We present the first analysis of a gene with functions relevant to this dimension of brain evolution. We show that NIN, a gene necessary for maintaining asymmetric, neurogenic divisions of radial glial cells (RGCs), evolved adaptively during anthropoid evolution. We explored how this selection relates to neural phenotypes and find a significant association between selection on NIN and neonatal brain size in catarrhines. Our analyses suggest a relationship with prenatal neurogenesis and identify the human data point as an outlier, possibly explained by postnatal changes in development on the human lineage. A similar pattern is found in platyrrhines, but the highly encephalized genus Cebus departs from the general trend. We further show that the evolution of NIN may be associated with variation in neuron number not explained by increases in surface area, a result consistent with NIN's role in neurogenic divisions of RGCs. Our combined results suggest a role for NIN in the evolution of cortical development. © 2012 The Authors. Genes, Brain and Behavior © 2012 Blackwell Publishing Ltd and International Behavioural and Neural Genetics Society.

  4. Multiple Evolutionary Selections Involved in Synonymous Codon Usages in the Streptococcus agalactiae Genome.

    Science.gov (United States)

    Ma, Yan-Ping; Ke, Hao; Liang, Zhi-Ling; Liu, Zhen-Xing; Hao, Le; Ma, Jiang-Yao; Li, Yu-Gu

    2016-02-24

    Streptococcus agalactiae is an important human and animal pathogen. To better understand the genetic features and evolution of S. agalactiae, multiple factors influencing synonymous codon usage patterns in S. agalactiae were analyzed in this study. A- and U-ending rich codons were used in S. agalactiae function genes through the overall codon usage analysis, indicating that Adenine (A)/Thymine (T) compositional constraints might contribute an important role to the synonymous codon usage pattern. The GC3% against the effective number of codon (ENC) value suggested that translational selection was the important factor for codon bias in the microorganism. Principal component analysis (PCA) showed that (i) mutational pressure was the most important factor in shaping codon usage of all open reading frames (ORFs) in the S. agalactiae genome; (ii) strand specific mutational bias was not capable of influencing the codon usage bias in the leading and lagging strands; and (iii) gene length was not the important factor in synonymous codon usage pattern in this organism. Additionally, the high correlation between tRNA adaptation index (tAI) value and codon adaptation index (CAI), frequency of optimal codons (Fop) value, reinforced the role of natural selection for efficient translation in S. agalactiae. Comparison of synonymous codon usage pattern between S. agalactiae and susceptible hosts (human and tilapia) showed that synonymous codon usage of S. agalactiae was independent of the synonymous codon usage of susceptible hosts. The study of codon usage in S. agalactiae may provide evidence about the molecular evolution of the bacterium and a greater understanding of evolutionary relationships between S. agalactiae and its hosts.

  5. Functional characterization of aromatic amino acid aminotransferase involved in 2-phenylethanol biosynthesis in isolated rose petal protoplasts.

    Science.gov (United States)

    Hirata, Hiroshi; Ohnishi, Toshiyuki; Ishida, Haruka; Tomida, Kensuke; Sakai, Miwa; Hara, Masakazu; Watanabe, Naoharu

    2012-03-15

    In rose flowers, 2-phenylethanol (2PE) is biosynthesized from l-phenylalanine (l-Phe) via phenylacetaldehyde (PAld) by the actions of two enzymes, pyridoxal-5'-phosphate (PLP)-dependent aromatic amino acid decarboxylase (AADC) and phenylacetaldehyde reductase (PAR). We here report that Rosa 'Yves Piaget' aromatic amino acid aminotransferase produced phenylpyruvic acid (PPA) from l-Phe in isolated petal protoplasts. We have cloned three full length cDNAs (RyAAAT1-3) of aromatic amino acid aminotransferase families based on rose EST database and homology regions. The RyAAATs enzymes were heterogeneously expressed in Escherichia coli and characterized biochemically. The recombinant RyAAAT3 showed the highest activity toward l-Phe in comparison with l-tryptophan, l-tyrosine, d-Phe, glycine, and l-alanine, and showed 9.7-fold higher activity with l-Phe rather than PPA as a substrate. RyAAAT3 had an optimal activity at pH 9 and at 45-55°C with α-ketoglutaric acid, and was found to be a PLP dependent enzyme based on the inhibition test using Carbidopa, an inhibitor of PLP-dependent enzymes. The transcript of RyAAAT3 was expressed in flowers as well as other organs of R. 'Yves Piaget'. RNAi suppression of RyAAAT3 decreased 2PE production, revealing the involvement of RyAAAT3 in 2PE biosynthesis in rose protoplasts and indicating that rose protoplasts have potentially two different 2PE biosynthetic pathways, the AADC route and the new route via PPA from l-Phe.

  6. Selective presynaptic insectotoxin (alpha-latroinsectotoxin) isolated from black widow spider venom.

    Science.gov (United States)

    Magazanik, L G; Fedorova, I M; Kovalevskaya, G I; Pashkov, V N; Bulgakov, O V; Grishin, E V

    1992-01-01

    A homogenous protein of 120,000 mol. wt isolated from black widow spider (Lactrodectus mactans tredecimguttatus) venom and referred to as alpha-latroinsectotoxin was highly potent (4 nM) in the induction of an increase of the frequency of miniature excitatory postsynaptic potentials in blowfly (Calliphora vicina) larvae neuromuscular preparations. In the frog nerve ending, however, even 50 nM alpha-latroinsectotoxin failed to affect transmitter release. Pretreatment of insect preparations with alpha-latrotoxin or frog preparations with alpha-latroinsectotoxin did not prevent the specific effect of consequent applications of alpha-latroinsectotoxin (insect) and alpha-latrotoxin (frog), respectively. The binding of labelled [125I]alpha-latroinsectotoxin to insect and [125I]alpha-latrotoxin to bovine membrane preparations was saturable and highly specific. The presynaptic effect, but not the binding of alpha-latroinsectotoxin, was dependent on the presence of divalent cations in the external medium. Mg2+ could readily substitute for Ca2+ and increase of transmitter release induced by alpha-latroinsectotoxin also occurred in Ca(2+)-free solutions. Pretreatment of preparations with 300 micrograms/ml concanavalin A completely abolished both the presynaptic effect of alpha-latroinsectotoxin and its binding to insect membrane preparations. Thus, the phenomenology of alpha-latroinsectotoxin action on insects resembles in general that described for the action of alpha-latrotoxin on vertebrates. The selectivity of alpha-latrotoxin and alpha-latroinsectotoxin seems to be due to differences in the structure of neurotoxin receptors in nerve endings of vertebrates and insects, although the mode of presynaptic action has a great deal in common.

  7. Characterization of Candida Species Isolated from cases of Vulvovaginitis in women referring to selected gynecological clinics

    Directory of Open Access Journals (Sweden)

    Batol Bonyadpour

    2016-10-01

    Full Text Available Background and aim: About 20% of non-pregnant women aged 15 to 55 harbour Candida albicans in the vagina .the aimed to determine the Characterization of Candida Species Isolated from women with Vulvovaginitis candidates (VVC of reproductive ages. Methods: this descriptive study was conducted on 280 of who were selected for gathering samples by Purposive sampling based on their history and characteristics of vaginal discharges in 2009. Among these patients, 105 ones were diagnosed with candidiasis. The data were collected using demographic information form and disease symptoms. the species were differentiated using germ tube test, chrome agar test, and chlamidospore test. Data analysis was performed in SPSS V.16, using Descriptive Statistics Results: the prevalence of candida vaginitis was 9.3%.105 samples obtained from patients.. Chlamidospore was detected in 54.3% of the corn meal agar media. Besides, in chrome agar test, 41.9% of the samples turned into green representing candida albicans. In germ tube test, on the other hand, 70.5% of the samples were candida albicans, while 29.5% were candida non-albicans. Overall, The frequency of the Candida albicans, Candida glabrata, Candida tropicalis and  the Candida Krusei were  66.6% , 219%  , 8.6% ,  and 2.9%, respectively. Conclusion: Candida albicans was the most common species leading to the Vulvovaginitis in patients with VCC while other species were at the secondary importance stages.Due to inaccurate diagnosis of the disease based on the clinical symptoms, fungal culture is recommended as a standard diagnostic method.

  8. Selection and characterization of Beauveria spp. isolates to control the broad mite Polyphagotarsonemus latus (Banks, 1904 (Acari: Tarsonemidae

    Directory of Open Access Journals (Sweden)

    C. C. Martins

    Full Text Available Abstract This study was performed under laboratory conditions to identify isolates of the fungus Beauveria spp. that can control Polyphagotarsonemus latus in the greenhouse and field. Thirty Beauveria spp. isolates were tested by spraying 1 mL conidia (1 × 108 conidia/mL on pepper leaf discs containing 15 mites. Evaluations were performed on the 3rd and 6th day post application by counting the number of dead mites. Vegetative growth and conidial production were measured from the selected isolates, and bioassays were conducted in the greenhouse on bean seedlings in plastic pots. The isolate Unioeste 53 was selected, and a conidial suspension (1 × 108 conidia/mL was applied with a backpack sprayer. The evaluation consisted of pre- and post-treatment counts of the number of live mites on ten leaflets in both the plots treated with the fungus and control plots, and the same procedure was followed for the field experiment. In the laboratory, the Unioeste 53 isolate resulted in total and confirmed mortality rates of 70% and 57.7%, respectively. In the greenhouse, the population decreased by 76.71% by the 16th day after application. In the field, the population decreased by 66% by the 12th day after application, demonstrating the potential of this fungus for mite management.

  9. Positive selection in development and growth rate regulation genes involved in species divergence of the genus Radix.

    Science.gov (United States)

    Feldmeyer, Barbara; Greshake, Bastian; Funke, Elisabeth; Ebersberger, Ingo; Pfenninger, Markus

    2015-08-19

    Life history traits like developmental time, age and size at maturity are directly related to fitness in all organisms and play a major role in adaptive evolution and speciation processes. Comparative genomic or transcriptomic approaches to identify positively selected genes involved in species divergence can help to generate hypotheses on the driving forces behind speciation. Here we use a bottom-up approach to investigate this hypothesis by comparative analysis of orthologous transcripts of four closely related European Radix species. Snails of the genus Radix occupy species specific distribution ranges with distinct climatic niches, indicating a potential for natural selection driven speciation based on ecological niche differentiation. We then inferred phylogenetic relationships among the four Radix species based on whole mt-genomes plus 23 nuclear loci. Three different tests to infer selection and changes in amino acid properties yielded a total of 134 genes with signatures of positive selection. The majority of these genes belonged to the functional gene ontology categories "reproduction" and "genitalia" with an overrepresentation of the functions "development" and "growth rate". We show here that Radix species divergence may be primarily enforced by selection on life history traits such as (larval-) development and growth rate. We thus hypothesise that life history differences may confer advantages under the according climate regimes, e.g., species occupying warmer and dryer habitats might have a fitness advantage with fast developing susceptible life stages, which are more tolerant to habitat desiccation.

  10. Global Variation of Human Papillomavirus Genotypes and Selected Genes Involved in Cervical Malignancies.

    Science.gov (United States)

    Husain, R S Akram; Ramakrishnan, V

    2015-01-01

    Carcinoma of the cervix is ranked second among the top 5 cancers affecting women globally. Parallel to other cancers, it is also a complex disease involving numerous factors such as human papillomavirus (HPV) infection followed by the activity of oncogenes and environmental factors. The incidence rate of the disease remains high in developing countries due to lack of awareness, followed by mass screening programs, various socioeconomic issues, and low usage of preventive vaccines. Over the past 3 decades, extensive research has taken place in cervical malignancy to elucidate the role of host genes in the pathogenesis of the disease, yet it remains one of the most prevalent diseases. It is imperative that recent genome-wide techniques be used to determine whether carcinogenesis of oncogenes is associated with cervical cancer at the molecular level and to translate that knowledge into developing diagnostic and therapeutic tools. The aim of this study was to discuss HPV predominance with their genotype distribution worldwide, and in India, as well as to discuss the newly identified oncogenes related to cervical cancer in current scenario. Using data from various databases and robust technologies, oncogenes associated with cervical malignancies were identified and are explained in concise manner. Due to the advent of recent technologies, new candidate genes are explored and can be used as precise biomarkers for screening and developing drug targets. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  11. Isolated Islands by Selective Local Oxidation (islo): a Silicon-On (soi) Technology for Nanoelectronic and Nanoelectromechanical Applications.

    Science.gov (United States)

    Arney, Susanne Christine

    The development of an advanced fully-integrated nanometer-scale isolation technology called the Isolated Islands of Substrate-Silicon by Selective Lateral Oxidation (ISLO) technology is reported. The versatility and applicability of the ISLO technology for diverse nanoelectronic and nanoelectromechanical devices and systems are described relative to the challenging issues of isolation and contacts. The basic ISLO structure is fabricated using electron beam lithography and standard VLSI reactive ion etching and oxidation processes. Single crystal silicon (SCS) islands 100-300-nm-wide, and 500 -2000-nm-tall are electrically and thermally isolated from the underlying substrate by selective lateral thermal oxidation at the base of the islands. Dislocation-free fully-isolated islands are obtained. Full-isolation of the basic ISLO structure depends on island linewidth, oxidation-masking film thicknesses, recess etch profile, and oxidation time and temperature. The extended ISLO technology provides 100-nm-wide, movable, suspended, high stiffness, low mass, SCS or SCS-dielectric-composite beam segments with integrated electrical contacts and metallization for high frequency (5-10 MHz) nanodynamic applications. Fixed or cantilevered beam segments are isolated from the underlying substrate -silicon by thermally grown oxide or an air-bridge. Wedge -pairs or tip-pairs vertically opposed across the isolation oxide or air-bridge have application to electron tunneling or field emission devices. A selectively-sharpened tip -above-a-tip structure is formed at the intersection of cantilevered beam segments. Vertical triple-tip and quadruple -tip structures are demonstrated. A new deep-submicron self-aligned sidewall source/drain, top-surface gate Thin -Film-Silicon-On-Insulator (TFSOI) MOSFET (ISLO FET) based on the inherently three-dimensional, non-planar ISLO structure is presented. Stress-related defect generation and dopant segregation during the oxidation, erosion of the high

  12. A Major Locus Controls a Genital Shape Difference Involved in Reproductive Isolation Between Drosophila yakuba and Drosophila santomea.

    Science.gov (United States)

    Peluffo, Alexandre E; Nuez, Isabelle; Debat, Vincent; Savisaar, Rosina; Stern, David L; Orgogozo, Virginie

    2015-10-28

    Rapid evolution of genitalia shape, a widespread phenomenon in animals with internal fertilization, offers the opportunity to dissect the genetic architecture of morphological evolution linked to sexual selection and speciation. Most quantitative trait loci (QTL) mapping studies of genitalia divergence have focused on Drosophila melanogaster and its three most closely related species, D. simulans, D. mauritiana, and D. sechellia, and have suggested that the genetic basis of genitalia evolution involves many loci. We report the first genetic study of male genitalia evolution between D. yakuba and D. santomea, two species of the D. melanogaster species subgroup. We focus on male ventral branches, which harm females during interspecific copulation. Using landmark-based geometric morphometrics, we characterized shape variation in parental species, F1 hybrids, and backcross progeny and show that the main axis of shape variation within the backcross population matches the interspecific variation between parental species. For genotyping, we developed a new molecular method to perform multiplexed shotgun genotyping (MSG), which allowed us to prepare genomic DNA libraries from 365 backcross individuals in a few days using little DNA. We detected only three QTL, one of which spans 2.7 Mb and exhibits a highly significant effect on shape variation that can be linked to the harmfulness of the ventral branches. We conclude that the genetic architecture of genitalia morphology divergence may not always be as complex as suggested by previous studies.

  13. PKCδ Inhibition Impairs Mammary Cancer Proliferative Capacity But Selects Cancer Stem Cells, Involving Autophagy.

    Science.gov (United States)

    Berardi, Damián E; Flumian, Carolina; Rodriguez, Cristina E; Bessone, María I Díaz; Cirigliano, Stefano M; Joffé, Elisa D Bal de Kier; Fiszman, Gabriel L; Urtreger, Alejandro J; Todaro, Laura B

    2016-03-01

    Protein kinase C (PKC) is a family of serine/threonine kinases that regulate diverse cellular functions including cell death, proliferation, and survival. Recent studies have reported that PKCδ, are involved in apoptosis or autophagy induction. In the present study we focused on how PKCδ regulates proliferation and cancer stem cell (CSC) properties of the hormone-independent mammary cancer cell line LM38-LP, using pharmacological and genetic approaches. We found that pharmacological inhibition of PKCδ, by Rottlerin treatment, impairs in vitro LM38-LP proliferation through cell cycle arrest, inducing the formation of cytoplasmic-vacuoles. Using immunofluorescence we confirmed that Rottlerin treatment induced the apparition of LC3 dots in cell cytoplasm, and increased autophagy flux. On the other side, the same treatment increased CSC growth rate and self-renewal. Furthermore, Rottlerin pre-treatment induced in CSC the development of a "grape-like" morphology when they are growing in 3D cultures (Matrigel), usually associated with a malignant phenotype, as well as an increase in the number of experimental lung metastasis when these cells were inoculated in vivo. The PKCδ knockdown, by RNA interference, induced autophagy and increased CSC number, indicating that these effects are indeed exerted through a PKCδ dependent pathway. Finally, the increase in the number of mammospheres could be reversed by a 3MA treatment, suggesting that autophagy mechanism is necessary for the increased of CSC self-renewal induced by PKCδ inhibition. Here we demonstrated that PKCδ activity exerts a dual role through the autophagy mechanism, decreasing proliferative capacity of mammary tumor cells but also regulating tumor stem cell self-renewal. © 2015 Wiley Periodicals, Inc.

  14. Radiographic implications of procedures involving cardiac implantable electronic devices (CIEDs – Selected aspects

    Directory of Open Access Journals (Sweden)

    Roman Steckiewicz

    2017-06-01

    Full Text Available Background: Some cardiac implantable electronic device (CIED implantation procedures require the use of X-rays, which is reflected by such parameters as total fluoroscopy time (TFT and dose-area product (DAP – defined as the absorbed dose multiplied by the area irradiated. Material and Methods: This retrospective study evaluated 522 CIED implantation (424 de novo and 98 device upgrade and new lead placement procedures in 176 women and 346 men (mean age 75±11 years over the period 2012–2015. The recorded procedure-related parameters TFT and DAP were evaluated in the subgroups specified below. The group of 424 de novo procedures included 203 pacemaker (PM and 171 implantable cardioverter-defibrillator (ICD implantation procedures, separately stratified by single-chamber and dual-chamber systems. Another subgroup of de novo procedures involved 50 cardiac resynchronization therapy (CRT devices. The evaluated parameters in the group of 98 upgrade procedures were compared between 2 subgroups: CRT only and combined PM and ICD implantation procedures. Results: We observed differences in TFT and DAP values between procedure types, with PM-related procedures showing the lowest, ICD – intermediate (with values for single-chamber considerably lower than those for dual-chamber systems and CRT implantation procedures – highest X-ray exposure. Upgrades to CRT were associated with 4 times higher TFT and DAP values in comparison to those during other upgrade procedures. Cardiac resynchronization therapy de novo implantation procedures and upgrades to CRT showed similar mean values of these evaluated parameters. Conclusions: Total fluoroscopy time and DAP values correlated progressively with CIED implantation procedure complexity, with CRT-related procedures showing the highest values of both parameters. Med Pr 2017;68(3:363–374

  15. Mechanisms involved in the selective transfer of long chain polyunsaturted fatty acids to the fetus

    Directory of Open Access Journals (Sweden)

    Alfonso eGil-Sánchez

    2011-09-01

    Full Text Available The concentration of long chain polyunsaturated fatty acid (LCPUFA in the fetal brain increases dramatically from the third trimester until 18 months of life. Several studies have shown an association between the percentage of maternal plasma docosahexaenoic acid (DHA during gestation and development of the cognitive functions in the neonate. Since only very low levels of LCPUFA are synthesized in the fetus and placenta, their primary source for the fetus is that of maternal origin. Both in vitro and human in vivo studies using labelled fatty acids have shown the preferential transfer of LCPUFA from the placenta to the fetus compared with other fatty acids, although the mechanisms involved are still uncertain. The placenta takes up circulating maternal non-esterified fatty acids (NEFA and fatty acids released mainly by maternal lipoprotein lipase and endothelial lipase. These NEFA may enter the cell by passive diffusion or by means of membrane carrier proteins. Once in the cytosol, NEFA bind to cytosolic fatty acid-binding proteins for transfer to the fetal circulation or can be oxidized within the trophoblasts and even re-esterified and stored in lipid droplets (LD. Although trophoblast cells are not specialized in lipid storage, LCPUFA may up-regulate peroxisome proliferator activated receptor-γ (PPARγ and hence the gene expression of fatty acid transport carriers, fatty acid acyl-CoA synthetases and adipophilin or other enzymes related with lipolysis, modifying their rate of placental transfer and metabolization. The placental transfer of LCPUFA during pregnancy seems to be a key factor in the neurological development of the fetus. Increased knowledge on the factors that modify placental transfer of fatty acids would contribute to our understanding of this complex process.

  16. The UV responses of bacterioneuston and bacterioplankton isolates depend on the physiological condition and involve a metabolic shift.

    Science.gov (United States)

    Santos, Ana L; Baptista, Inês; Lopes, Sílvia; Henriques, Isabel; Gomes, Newton C M; Almeida, Adelaide; Correia, António; Cunha, Angela

    2012-06-01

    Bacteria from the surface microlayer (bacterioneuston) and underlying waters (bacterioplankton) were isolated upon exposure to UV-B radiation, and their individual UV sensitivity in terms of CFU numbers, activity (leucine and thymidine incorporation), sole-carbon source use profiles, repair potential (light-dependent and independent), and photoadaptation potential, under different physiological conditions, was compared. Colony counts were 11.5-16.2% more reduced by UV-B exposure in bacterioplankton isolates (P < 0.05). Inhibition of leucine incorporation in bacterioneuston isolates was 10.9-11.5% higher than in bacterioplankton (P < 0.05). These effects were accompanied by a shift in sole-carbon source use profiles, assessed with Biolog(®) EcoPlates, with a reduction in consumption of amines and amino acids and increased use of polymers, particularly in bacterioneuston isolates. Recovery under starvation was generally enhanced compared with nourished conditions, especially in bacterioneuston isolates. Overall, only insignificant increases in the induction of antibiotic resistant mutant phenotypes (Rif(R) and Nal(R) ) were observed. In general, a potential for photoadaptation could not be detected among the tested isolates. These results indicate that UV effects on bacteria are influenced by their physiological condition and are accompanied by a shift in metabolic profiles, more significant in bacterioneuston isolates, suggesting the presence of bacterial strains adapted to high UV levels in the SML.

  17. Isolation and selection of fluorescent pseudomonads based on multiple plant growth promotion traits and siderotyping

    Directory of Open Access Journals (Sweden)

    Jayamohan Subramanian

    2014-09-01

    Full Text Available Fluorescent pseudomonads, acclaimed plant associated bacterial group, are well-known plant growth promoting-biocontrol agents in rhizosphere arena. In this study, 144 fluorescent pseudomonad isolates from rhizosphere soil samples were screened with King's medium B supplemented with 8-hydroxyquinoline (8-HQ chelator and comprehensively profiled for plant growth promotion viz., production of indole acetic acid (IAA, siderophore, ammonia, hydrogen cyanide, motility, phosphate solubilization, root growth promotion, and biofilm forming ability, along with two known control strains of pseudomonads. Iron and IAA regulated secondary metabolite siderophore production were investigated quantitatively. All isolates were positive for ammonia production and motility; 46% isolates were positive for hydrogen cyanide, 44% shown positivity for phosphate solubilization, and 40% isolates for siderophore production. Siderotyping showed production of hydroxamate type of siderophores which are known to be more efficient biocontrol agents. All isolates stimulated root growth to varying extent and had potentiality to form biofilms, a critical constituent for survival on different environments. Forty-two isolates of pseudomonads showed antagonistic behavior against the deleterious fungal pathogen Fusarium oxysporum (MTCC1755. Based on the above observations and statistical analysis, 11 isolates were shortlisted for further scrutiny. The study of biogeographic correlation and secondary metabolite profiling in association with plant growth promotion focalizes significant assessment on the behavior and antagonistic action, which probably brings out a competent biocontrol agent in a sustainable eco-friendly dimension.

  18. Disease Risk Assessments Involving Companion Animals: an Overview for 15 Selected Pathogens Taking a European Perspective.

    Science.gov (United States)

    Rijks, J M; Cito, F; Cunningham, A A; Rantsios, A T; Giovannini, A

    2016-07-01

    Prioritization of companion animal transmissible diseases was performed by the Companion Animals multisectoriaL interprofessionaL Interdisciplinary Strategic Think tank On zoonoses (CALLISTO) project. The project considered diseases occurring in domesticated species commonly kept as pets, such as dogs and cats, but also included diseases occurring in captive wild animals and production animal species. The prioritization process led to the selection of 15 diseases of prime public health relevance, agricultural economic importance, or both. An analysis was made of the current knowledge on the risk of occurrence and transmission of these diseases among companion animals, and from companion animals to man (zoonoses) or to livestock. The literature was scanned for risk assessments for these diseases. Studies were classified as import risk assessments (IRAs) or risk factor analyses (RFAs) in endemic areas. For those pathogens that are absent from Europe, only IRAs were considered; for pathogens present throughout Europe, only RFAs were considered. IRAs were identified for seven of the eight diseases totally or partially absent from Europe. IRAs for classical rabies and alveolar echinococcosis found an increased risk for introduction of the pathogen into officially disease-free areas as a consequence of abandoning national rules and adopting the harmonized EU rules for pet travel. IRAs for leishmaniosis focused on risk associated with the presence of persistently infected dogs in new geographical areas, taking into consideration the risk of disease establishment should a competent vector arise. IRAs for Crimean-Congo haemorrhagic fever and West Nile fever indicated that the likelihood of introduction via companion animals was low. IRAs for bluetongue paid no attention to the risk of introduction via companion animals, which was also the case for IRAs for foot-and-mouth disease, the only disease considered to be absent from Europe. RFAs dealing with the risk factors for

  19. Factors associated with crashes involving taxi owners and non-owners: A case of moral hazard and adverse selection?

    Science.gov (United States)

    Tay, Richard; Choi, Jaisung

    2016-02-01

    Taxis experience a higher risk of a motor vehicle crash partly because of their much higher levels of exposure on the roads. Although several studies have been conducted to examine the factors associated with the frequency and severity of taxi collisions, little research has been conducted to examine the differences in the factors associated with owner taxis and non-owner taxis. This study finds that collisions involving non-owners are more likely to be associated with poor or risky driving behaviors than collisions involving taxi vehicle owners. This result is consistent with the economic principles of moral hazard and adverse selection. Hence, policy makers responsible for traffic safety, taxi regulation or taxi operations should consider measures to reduce these market inefficiencies and improve the safety of not only taxi drivers but all road users.

  20. Clonal diversity in Giardia duodenalis isolates from Thailand: evidences for intragenic recombination and purifying selection at the beta giardin locus.

    Science.gov (United States)

    Kosuwin, Rattiporn; Putaporntip, Chaturong; Pattanawong, Urassaya; Jongwutiwes, Somchai

    2010-01-01

    The beta giardin locus of Giardia duodenalis encodes a structural component of ventral disc and exhibits sequence variation among isolates rendering it a useful marker for genotypic analysis. To determine the distribution of genotypes of G. duodenalis in Thailand and to explore the extent of sequence variation in this locus, we deployed the PCR-RFLP method and sequence analysis of recombinant subclones from 30 clinical isolates. In total, assemblage B was more prevalent than assemblage A. Sequence analysis revealed that 13 isolates had clonal mixtures, comprising three to five distinct sequences per isolate. Nucleotide diversity of assemblage B was greater than that of assemblage A. A striking transitional bias was noted at the first and the third positions of codons in both assemblages; however, they differed in the patterns of nucleotide diversity at 0-fold and 4-fold-degenerate sites. Most amino acid exchanges were conservative in terms of polarity, charge and volume. Both assemblages have evolved under purifying selection as evidenced by a significantly greater number of mean synonymous substitutions per synonymous site (d(S)) than that of nonsynonymous substitutions per nonsynonymous site (d(N)) as well as significant negative Tajima's D values and its related statistics. The significant negative Tajima's D test at nonsynonymous sites further suggests that elimination of slightly deleterious mutations at these sites by purifying selection is ongoing as predicted in the nearly neutral theory. Furthermore, a minimum number of seven recombination sites was detected by the four gamete test in assemblage B, consistent with previous reports on meiotic recombination in G. duodenalis. Therefore, accurate subassemblage assignment of clinical isolates that has practical consequence for disease control could be complicated by the presence of intra-isolate clonal diversity and interallelic recombination.

  1. Genetic architecture of a reinforced, postmating, reproductive isolation barrier between Neurospora species indicates evolution via natural selection.

    Directory of Open Access Journals (Sweden)

    Elizabeth Turner

    2011-08-01

    Full Text Available A role for natural selection in reinforcing premating barriers is recognized, but selection for reinforcement of postmating barriers remains controversial. Organisms lacking evolvable premating barriers can theoretically reinforce postmating isolation, but only under restrictive conditions: parental investment in hybrid progeny must inhibit subsequent reproduction, and selected postmating barriers must restore parents' capacity to reproduce successfully. We show that reinforced postmating isolation markedly increases maternal fitness in the fungus Neurospora crassa, and we detect the evolutionary genetic signature of natural selection by quantitative trait locus (QTL analysis of the reinforced barrier. Hybrid progeny of N. crassa and N. intermedia are highly inviable. Fertilization by local N. intermedia results in early abortion of hybrid fruitbodies, and we show that abortion is adaptive because only aborted maternal colonies remain fully receptive to future reproduction. In the first QTL analysis of postmating reinforcement in microbial eukaryotes, we identify 11 loci for abortive hybrid fruitbody development, including three major QTLs that together explain 30% of trait variance. One of the major QTLs and six QTLs of lesser effect are found on the mating-type determining chromosome of Neurospora. Several reinforcement QTLs are flanked by genetic markers showing either segregation distortion or non-random associations with alleles at other loci in a cross between N. crassa of different clades, suggesting that the loci also are associated with local effects on same-species reproduction. Statistical analysis of the allelic effects distribution for abortive hybrid fruitbody development indicates its evolution occurred under positive selection. Our results strongly support a role for natural selection in the evolution of reinforced postmating isolation in N. crassa.

  2. Molecular characterisation of Sporothrix schenckii isolates from humans and cats involved in the sporotrichosis epidemic in Rio de Janeiro, Brazil

    Directory of Open Access Journals (Sweden)

    Rosani Santos Reis

    2009-08-01

    Full Text Available An epidemic of sporotrichosis, a subcutaneous mycosis caused by the fungus Sporothrix schenckii, is ongoing in Rio de Janeiro, Brazil, in which cases of human infection are related to exposure to cats. In an attempt to demonstrate the zoonotic character of this epidemic using molecular methodology, we characterised by DNA-based typing methods 19 human and 25 animal S. schenckii isolates from the epidemic, as well as two control strains. To analyse the isolates, the random amplified polymorphic DNA (RAPD technique was performed using three different primers, together with DNA fingerprinting using the minisatellite derived from the wild-type phage M13 core-sequence. The analyses generated amplicons with considerable polymorphism. Although isolates exhibited high levels of genetic relatedness, they could be clustered into 5-10 genotypes. The RAPD profiles of epidemic S. schenckii isolates could be distinguished from that of the United States isolate, displaying 20% similarity to each primer and 60% when amplified with the M13 primer. DNA fingerprinting of S. schenckii isolated from the nails (42.8% and the oral cavities (66% of cats were identical to related human samples, suggesting that there is a common infection source for animals and humans in this epidemic. It is clear that cats act as a vehicle for dissemination of S. schenckii.

  3. Comparative evaluation of five Beauveria isolates for housefly (Musca domestica L.) control and growth optimization of selected strain.

    Science.gov (United States)

    Mishra, Sapna; Malik, Anushree

    2012-11-01

    Pathogenic potential of five native Beauveria isolates was assessed against housefly adult and larvae in laboratory bioassays. Beauveria isolate Beauveria bassiana HQ917687 showed highest virulence with 72.3 and 100 % mortality of larvae and adults of Musca domestica, respectively. Other Beauveria isolates caused 36-52 % housefly larval mortality while the adult mortalities varied between 72 and 82 %. B. bassiana HQ917687 also showed the fastest killing activity with LT(50) of 4 days (for larvae) and 3 days (for adults). This isolate showing highest virulence was selected for its growth optimization in terms of biomass and spore production using response surface methodology. The optimum value of temperature, yeast extract, and pH for maximum biomass and spore production was predicted as 27 °C, 5.00 g/l, and 6.75, respectively. Temperature was found to be the most critical factor influencing biomass and spore yield of the fungus and even nullified the effects of other factors at sufficiently higher value. The results obtained in this study depict the significance of appropriate strain selection and process parameter optimization in order to facilitate mass production of biocontrol agents.

  4. Determination of the influence of substrate concentration on enzyme selectivity using whey protein Isolate and Bacillus licheniformis protease.

    Science.gov (United States)

    Butré, Claire I; Sforza, Stefano; Gruppen, Harry; Wierenga, Peter A

    2014-10-22

    Increasing substrate concentration during enzymatic protein hydrolysis results in a decrease in hydrolysis rate. To test if changes in the mechanism of hydrolysis also occur, the enzyme selectivity was determined. The selectivity is defined quantitatively as the relative rate of hydrolysis of each cleavage site in the protein. It was determined from the identification and quantification of the peptides present in the hydrolysates. Solutions of 0.1-10% (w/v) whey protein isolate (WPI) were hydrolyzed by Bacillus licheniformis protease at constant enzyme-to-substrate ratio. The cleavage sites were divided into five groups, from very high (>10%) to very low selectivity (protein concentrations. This finding shows that both the rate of hydrolysis and the enzyme selectivity were influenced by the substrate concentration.

  5. Isolation and selection of plant growth-promoting bacteria associated with sugarcane

    Directory of Open Access Journals (Sweden)

    Ariana Alves Rodrigues

    2016-06-01

    Full Text Available Microorganisms play a vital role in maintaining soil fertility and plant health. They can act as biofertilizers and increase the resistance to biotic and abiotic stress. This study aimed at isolating and characterizing plant growth-promoting bacteria associated with sugarcane, as well as assessing their ability to promote plant growth. Endophytic bacteria from leaf, stem, root and rhizosphere were isolated from the RB 867515 commercial sugarcane variety and screened for indole acetic acid (IAA production, ability to solubilize phosphate, fix nitrogen and produce hydrogen cyanide (HCN, ammonia and the enzymes pectinase, cellulase and chitinase. A total of 136 bacteria were isolated, with 83 of them presenting some plant growth mechanism: 47 % phosphate solubilizers, 26 % nitrogen fixers and 57 % producing IAA, 0.7 % HCN and chitinase, 45 % ammonia, 30 % cellulose and 8 % pectinase. The seven best isolates were tested for their ability to promote plant growth in maize. The isolates tested for plant growth promotion belong to the Enterobacteriaceae family and the Klebsiella, Enterobacter and Pantoea genera. Five isolates promoted plant growth in greenhouse experiments, showing potential as biofertilizers.

  6. Temporally isolated lineages of Pink salmon reveal unique signatures of selection on distinct pools of standing genetic variation

    DEFF Research Database (Denmark)

    Limborg, Morten; Waples, R.K.; Seeb, J.E.

    2014-01-01

    A species’ genetic diversity bears the marks of evolutionary processes that have occurred throughout its history. However, robust detection of selection in wild populations is difficult and often impeded by lack of replicate tests. Here, we investigate selection in pink salmon (Oncorhynchus...... gorbuscha) using genome scans coupled with inference from a haploid-assisted linkage map. Pink salmon have a strict 2-year semelparous life history which has resulted in temporally isolated (allochronic) lineages that remain sympatric through sharing of spawning habitats in alternate years. The lineages...

  7. Selected Lactobacillus strains isolated from sugary and milk kefir reduce Salmonella infection of epithelial cells in vitro.

    Science.gov (United States)

    Zavala, L; Golowczyc, M A; van Hoorde, K; Medrano, M; Huys, G; Vandamme, P; Abraham, A G

    2016-09-01

    The isolation of potentially probiotic strains and the subsequent study of their properties are very important steps to gain insight in the health benefits ascribed to sugary and milk kefir. The aim of the present study was to characterise fifteen Lactobacillus strains isolated from these beverages by determining some surface properties and their ability to antagonise enterocyte cell damage after Salmonella infection in vitro. Lactobacillus surface properties were determined by hydrophobicity, autoaggregation, and coaggregation assays with Salmonella. In addition, lactobacilli adhesion to Caco-2/TC-7 cells and the effect on Salmonella invasion were evaluated. Finally, the disassembly of F-actin cytoskeleton on intestinal epithelial cells was assayed in vitro when Salmonella infection was performed in the presence of selected Lactobacillus strains. Ten out of the 15 strains showed a high adhesion capacity to Caco-2/TC-7 cells. Most of the strains were hydrophilic and non-autoaggregating. Strains isolated from sugary kefir were non-coaggregating with Salmonella, while strains Lactobacillus paracasei CIDCA 83120, 83121, 83123, 83124, 8339, 83102 isolated from milk kefir were able to coaggregate after 1 h. L. paracasei CIDCA 8339 and Lactobacillus kefiri CIDCA 83102 were able to diminish Salmonella invasion to the enterocytes. An antagonistic effect on cytoskeleton disruption elicited by the pathogen was also demonstrated. Our results suggest that both strains isolated from milk kefir could be considered as appropriate probiotic candidates.

  8. Comparação da eficiência dos caldos de enriquecimento seletivo no isolamento de Salmonella Dublin Comparison of the efficiency of selective enrichment broths for Salmonella Dublin isolation

    Directory of Open Access Journals (Sweden)

    D.G. Silva

    2008-06-01

    Full Text Available The aim of this study was to compare three different selective enrichment broths: Rappaport-Vassiliadis (RV, selenite cystine (SC and Muller-Kauffmann tetrathionate (MKT for Salmonella Dublin isolation from faecal samples of calf experimentally infected. The bacteriological procedure involved pre-enrichment stages in Hajna-GN broth (only for the samples inoculated in RV broth, selective enrichment, culture in modified brilliant green agar (BGA, presumptive biochemistry tests (using triple-sugar-iron agar and lysine-agar and slide agglutination test with poli-O and poli-H Salmonella antiserum. The effects of enrichment temperatures using RV broth were also evaluated (37ºC and 42ºC. SC broth was significantly more efficient in the isolation of Salmonella Dublin (P<0,05, whereas RV broth incubated at 42ºC had a lower efficiency in the microbiological isolation.

  9. Selection of potential probiotic Enterococcus faecium isolated from Portuguese fermented food.

    Science.gov (United States)

    Barbosa, Joana; Borges, Sandra; Teixeira, Paula

    2014-11-17

    Four Enterococcus faecium strains isolated from fermented products were evaluated for potential use as probiotic strains. In addition to efaAfm gene, commonly found in E. faecium food isolates, none of the isolates possessed virulence genes and none had positive reactions for the production of tyramine, histamine, putrescine and cadaverine in the screening medium used. All of these four isolates proved to be resistant to 65 °C. E. faecium 119 did not show antimicrobial activity against any of the target bacteria investigated. E. faecium 85 and 101 inhibited Listeria innocua and E. faecium DSMZ 13590. The strain E. faecium 120 inhibited seven target bacteria (Listeria monocytogenes 7946, L. monocytogenes 7947, L. innocua 2030c, L. innocua NCTC 11286, E. faecium DSMZ 13590, Enterococcus faecalis ATCC 29212 and Staphylococcus aureus ATCC 29213) and was chosen as the representative to assess the ability to survive gastrointestinal tract passage simulation, as well as the protective role of two food matrices (skim milk and Alheira) during its passage. For both matrices used, no significant differences (pdetection limit of the enumeration technique by the end of the two digestions, in contrast to the Alheira matrix, for which isolate 120 showed a reduction of only ca. 1 log CFU/ml. The E. faecium strain 120 was shown to be a potential candidate for further investigations as a potential probiotic culture.

  10. Antimicrobial susceptibility survey of pathogens isolated from selected patients in Northern Italy

    Directory of Open Access Journals (Sweden)

    Elisabetta Maioli

    2005-03-01

    Full Text Available The Clinical Microbiology Laboratory of the University of Genoa participated, during the year 2003, in an international antimicrobial surveillance program.The collection of isolates was done according to the site of infection and/or type of patient. Four hundred twenty (420 clinical isolates were analyzed during this year and the frequencies of the different pathogens were investigated. A reference centre carried out susceptibility tests. Oxacillin-resistant Staphylococcus aureus represented 47.6% of all S. aureus isolates from blood stream infections and 33.3% of all S. aureus isolated from skin and soft tissue infections in hospitalised patients.These strains showed resistance to most of the antimicrobial agents evaluated, except vancomycin, teicoplanin, quinupristin/dalfopristin and linezolid which registered 100% of susceptibility. Some isolates from blood stream infections such as E. coli demonstrated resistance to ciprofloxacin (23.3%, levofloxacin (20%, and gatifloxacin (16.6%, and Klebsiella pneumoniae was resistant (18% to all fluoroquinolones tested. Pseudomonas aeruginosa manifested resistance to ciprofloxacin (16.6%, while 27.7% of these strains were resistant both to levofloxacin and gatifloxacin. All the Enterobacter cloacae isolated from blood were susceptible to ciprofloxacin, levofloxacin and gatifloxacin. Haemophilus influenzae and Moraxella catarrhalis collected from community-acquired respiratory tract infections were all inhibited by ciprofloxacin, levofloxacin and gatifloxacin. E. coli isolated from urinary tract infections in hospitalised patients were resistant to ciprofloxacin, levofloxacin and gatifloxacin (2.7%. All Salmonella spp. collected from samples of patients affected by infections of the gastro-intestinal tract were susceptible to all fluoroquinolones. Penicillin resistance in Streptococcus pneumoniae was found in 21.4% of isolates from patients with respiratory tract infections. Fluoroquinolone resistance was

  11. Isolation and selection of new astaxanthin producing strains of Xanthophyllomyces dendrorhous.

    Science.gov (United States)

    Libkind, Diego; Moliné, Martín; Tognetti, Celia

    2012-01-01

    Astaxanthin is a xanthophyll pigment of high economic value for its use as a feeding component in aquaculture. Xanthophyllomyces dendrorhous is a basidiomycetous fungi able to synthesize astaxanthin as its major carotenoid, and the only known yeast species bearing the capability to produce this type of carotenoid. Recently, the habitat and intraspecific variability of this species have been found to be wider than previously expected, encouraging the search for new wild strains with potential biotechnological applications. Here we describe effective procedures for isolation of X. dendrorhous from environmental samples, accurate identification of the strains, analysis of their astaxanthin content, and proper conservation of the isolates.

  12. Comparison of Efflux Pump Involvement in Antibiotic Resistance Among Pseudomonas aeruginosa Isolates of Burn and Non-Burn Patients

    Directory of Open Access Journals (Sweden)

    Azimi

    2016-05-01

    Full Text Available Background Pseudomonas aeruginosa is an important cause of hospital-acquired infections that can create serious problem for patients and physicians. Many factors are associated with the antibiotic resistance of P. aeruginosa, such as efflux pumps. Objectives The aim of this study was the phenotypic and molecular detection of efflux pumps in our clinical P. aeruginosa isolates in a comparison between burn and non-burn specimens. Materials and Methods In this study, clinical strains of P. aeruginosa were collected from burn and non-burn specimens from April - July 2013. Antibiotic susceptibility testing of the isolates was performed after biochemical and molecular identification. The minimum inhibitory concentration (MIC of imipenem, cefepime, gentamycin, and ciprofloxacin, with and without carbonyl cyanide 3-chlorophenylhydrazone (CCCP, was determined for phenotypic detection of efflux pumps. Results Our results confirmed 203 and 60 P. aeruginosa isolates from burn and non-burn specimens, respectively. The most antibiotic resistance was observed against tobramycin in both group of specimens, and no resistance was seen to colistin. Phenotypic detection of efflux pumps was determined to correlate to a > 4-fold decrease in the MICs of the tested antibiotics with CCCP compared to without CCCP in 57 strains. Conclusions High-level antibiotic resistance can occur as a result of multidrug efflux pumps combined with other mechanisms of resistance. However, the association between over-expression of these genes and highly resistant clinical isolates cannot be ignored.

  13. Positional Cloning in Xp22 : towards the isolation of the gene involved in X-linked retinoschisis

    NARCIS (Netherlands)

    Vosse, Esther van de

    1998-01-01

    The study was aimed at the positional cloning of disease genes in Xp22.1-p22.2. To this end a YAC contig covering this region was constructed. To identify candidate genes for the diseases localised in this region exon trapping was applied. Several novel transcripts were isolated from the region, of

  14. Molecular Analysis of the embCAB Locus and embR Gene Involved in Ethambutol Resistance in Clinical Isolates of Mycobacterium tuberculosis in France.

    Science.gov (United States)

    Brossier, Florence; Sougakoff, Wladimir; Bernard, Christine; Petrou, Matthieu; Adeyema, Karine; Pham, Anne; Amy de la Breteque, Diane; Vallet, Marine; Jarlier, Vincent; Sola, Christophe; Veziris, Nicolas

    2015-08-01

    Modification of codon 306 in embB is regarded as the main mechanism leading to ethambutol (ETB) resistance in clinical isolates of Mycobacterium tuberculosis. However, numerous mutations elsewhere in the embCAB locus and in embR, a putative transcriptional activator of this locus, have been reported to be involved in ETB resistance. Here, we investigated the diversity of nucleotide variations observed in embCAB and embR in M. tuberculosis complex isolates from France. These regions were sequenced in 71 ETB-resistant (ETB-R) and 60 ETB-susceptible (ETB-S) clinical isolates of known phylogenetic lineages. The 131 isolates had 12 mutations corresponding to phylogenetic markers. Among the 60 ETB-S isolates, only 3 (5%) had nonsynonymous mutations that were not phylogenetic markers. Among the 71 ETB-R isolates, 98% had mutations in embCAB that likely contribute to ETB resistance: 70% had mutations located in embB codon 306, 406, or 497; 13% had mutations located outside these three positions between codons 296 and 426; and 15% had mutations corresponding to mutations in the embC-embA intergenic region. We found a strong association between resistance to ETB and the presence of mutations in embB and the embC-embA intergenic region (P embA were not involved in ETB resistance, and no mutation was detected in embR. These results strongly suggest that the sensitivity of diagnostic assays for detecting ETB resistance based on testing of embB codon 306 can be increased by testing of the embB region between codons 296 and 497 and by including the embC-embA intergenic region between positions -8 and -21.

  15. A hidden Markov model to identify and adjust for selection bias: an example involving mixed migration strategies.

    Science.gov (United States)

    Fieberg, John R; Conn, Paul B

    2014-05-01

    An important assumption in observational studies is that sampled individuals are representative of some larger study population. Yet, this assumption is often unrealistic. Notable examples include online public-opinion polls, publication biases associated with statistically significant results, and in ecology, telemetry studies with significant habitat-induced probabilities of missed locations. This problem can be overcome by modeling selection probabilities simultaneously with other predictor-response relationships or by weighting observations by inverse selection probabilities. We illustrate the problem and a solution when modeling mixed migration strategies of northern white-tailed deer (Odocoileus virginianus). Captures occur on winter yards where deer migrate in response to changing environmental conditions. Yet, not all deer migrate in all years, and captures during mild years are more likely to target deer that migrate every year (i.e., obligate migrators). Characterizing deer as conditional or obligate migrators is also challenging unless deer are observed for many years and under a variety of winter conditions. We developed a hidden Markov model where the probability of capture depends on each individual's migration strategy (conditional versus obligate migrator), a partially latent variable that depends on winter severity in the year of capture. In a 15-year study, involving 168 white-tailed deer, the estimated probability of migrating for conditional migrators increased nonlinearly with an index of winter severity. We estimated a higher proportion of obligates in the study cohort than in the population, except during a span of 3 years surrounding back-to-back severe winters. These results support the hypothesis that selection biases occur as a result of capturing deer on winter yards, with the magnitude of bias depending on the severity of winter weather. Hidden Markov models offer an attractive framework for addressing selection biases due to their

  16. Hydrologic connectivity between geographically isolated wetlands and surface water systems: A review of select modeling methods

    Science.gov (United States)

    Heather E. Golden; Charles R. Lane; Devendra M. Amatya; Karl W. Bandilla; Hadas Raanan Kiperwas Kiperwas; Christopher D. Knightes; Herbert. Ssegane

    2014-01-01

    Geographically isolated wetlands (GIW), depressional landscape features entirely surrounded by upland areas, provide a wide range of ecological functions and ecosystem services for human well-being. Current and future ecosystem management and decision-making rely on a solid scientific understanding of how hydrologic processes affect these important GIW services and...

  17. Isolation and in vitro selection of actinomycetes strains as potential probiotics for aquaculture

    Directory of Open Access Journals (Sweden)

    Milagro García Bernal

    2015-02-01

    Full Text Available Aim: This study was designed to describe a series of in vitro tests that may aid the discovery of probiotic strains from actinomycetes. Materials and Methods: Actinomycetes were isolated from marine sediments using four different isolation media, followed by antimicrobial activity and toxicity assessment by the agar diffusion method and the hemolysis of human blood cells, respectively. Extracellular enzymatic production was monitored by the hydrolysis of proteins, lipids and carbohydrates. Tolerance to different pH values and salt concentrations was also determined, followed by hydrophobicity analysis and genetic identification of the most promising strains. Results: Five out of 31 isolated strains showed antimicrobial activity against three Vibrio species. Three non-hemolytic strains (N7, RL8 and V4 among these active isolates yielded positive results in hydrophobicity tests and exhibited good growth at salt concentrations ranging from 0% to 10%, except strain RL8, which required a salt concentration >0.6%. Although these strains did not grow at pH<3, they showed different enzymatic activities. Phylogenetic analysis revealed that strains N7 and V4 have more than 99% identity with several Streptomyces species, whereas the closest matches to strain RL8 are Streptomyces panacagri and Streptomyces flocculus, with 98% and 98.2% similarity, respectively. Conclusion: Three actinomycetes strains showing probiotic-like properties were discovered using several in vitro tests that can be easily implemented in different institutions around the world.

  18. Isolation and in vitro selection of actinomycetes strains as potential probiotics for aquaculture

    Science.gov (United States)

    Bernal, Milagro García; Campa-Córdova, Ángel Isidro; Saucedo, Pedro Enrique; González, Marlen Casanova; Marrero, Ricardo Medina; Mazón-Suástegui, José Manuel

    2015-01-01

    Aim: This study was designed to describe a series of in vitro tests that may aid the discovery of probiotic strains from actinomycetes. Materials and Methods: Actinomycetes were isolated from marine sediments using four different isolation media, followed by antimicrobial activity and toxicity assessment by the agar diffusion method and the hemolysis of human blood cells, respectively. Extracellular enzymatic production was monitored by the hydrolysis of proteins, lipids and carbohydrates. Tolerance to different pH values and salt concentrations was also determined, followed by hydrophobicity analysis and genetic identification of the most promising strains. Results: Five out of 31 isolated strains showed antimicrobial activity against three Vibrio species. Three non-hemolytic strains (N7, RL8 and V4) among these active isolates yielded positive results in hydrophobicity tests and exhibited good growth at salt concentrations ranging from 0% to 10%, except strain RL8, which required a salt concentration >0.6%. Although these strains did not grow at pH<3, they showed different enzymatic activities. Phylogenetic analysis revealed that strains N7 and V4 have more than 99% identity with several Streptomyces species, whereas the closest matches to strain RL8 are Streptomyces panacagri and Streptomyces flocculus, with 98% and 98.2% similarity, respectively. Conclusion: Three actinomycetes strains showing probiotic-like properties were discovered using several in vitro tests that can be easily implemented in different institutions around the world. PMID:27047067

  19. Selective loss of chemokine receptor expression on leukocytes after cell isolation.

    Directory of Open Access Journals (Sweden)

    Juan C Nieto

    Full Text Available Chemokine receptors are distinctively exposed on cells to characterize their migration pattern. However, little is known about factors that may regulate their expression. To determine the optimal conditions for an accurate analysis of chemokine receptors, we compared the expression of CCR2, CCR4, CCR5, CCR6, CXCR3 and CXCR4 on different leukocyte subsets using whole blood (WB plus erythrocyte lysis and density gradient isolation (Ficoll. Most WB monocytes were CCR2+ (93.5 ± 2.9% whereas 32.8 ± 6.0% of monocytes from Ficoll-PBMC expressed CCR2 (p<0.001. Significant reductions of CCR6 and CXCR3 on monocytes were also observed after Ficoll isolation (WB: 46.4 ± 7.5% and 57.1 ± 5.5%; Ficoll: 29.5 ± 2.2% and 5.4 ± 4.3% respectively (p<0.01. Although comparable percentages of WB and Ficoll-PBMC monocytes expressed CCR4, CCR5 and CXCR4, Ficoll isolation significantly reduced the levels of CXCR4 (WB: MFI 5 ± 0.4 and Ficoll: MFI 3.3 ± 0.1 (p<0.05. Similarly to monocytes, CCR2, CXCR3 and CXCR4 were also reduced on lymphocytes. In addition, Ficoll isolation significantly reduced the percentage of CCR4 positive lymphocytes (WB: 90.2 ± 4.5% and Ficoll: 55 ± 4.1% (p<0.01. The loss of expression of chemokine receptors after isolation of monocytes was not dependent on either the anticoagulant or the density gradient method. It was irreversible and could not be restored by LPS activation or in vitro macrophage differentiation. Experiments tagged with anti-CCR2 antibodies prior to density gradient isolation demonstrated that Ficoll internalized chemokine receptors. The method for cell isolation may alter not only the expression of certain chemokine receptors but also the respective functional migration assay. The final choice to analyze their expression should therefore depend on the receptor to be measured.

  20. Phylodynamic analysis of clinical and environmental Vibrio cholerae isolates from Haiti reveals diversification driven by positive selection.

    Science.gov (United States)

    Azarian, Taj; Ali, Afsar; Johnson, Judith A; Mohr, David; Prosperi, Mattia; Veras, Nazle M; Jubair, Mohammed; Strickland, Samantha L; Rashid, Mohammad H; Alam, Meer T; Weppelmann, Thomas A; Katz, Lee S; Tarr, Cheryl L; Colwell, Rita R; Morris, J Glenn; Salemi, Marco

    2014-12-23

    Phylodynamic analysis of genome-wide single-nucleotide polymorphism (SNP) data is a powerful tool to investigate underlying evolutionary processes of bacterial epidemics. The method was applied to investigate a collection of 65 clinical and environmental isolates of Vibrio cholerae from Haiti collected between 2010 and 2012. Characterization of isolates recovered from environmental samples identified a total of four toxigenic V. cholerae O1 isolates, four non-O1/O139 isolates, and a novel nontoxigenic V. cholerae O1 isolate with the classical tcpA gene. Phylogenies of strains were inferred from genome-wide SNPs using coalescent-based demographic models within a Bayesian framework. A close phylogenetic relationship between clinical and environmental toxigenic V. cholerae O1 strains was observed. As cholera spread throughout Haiti between October 2010 and August 2012, the population size initially increased and then fluctuated over time. Selection analysis along internal branches of the phylogeny showed a steady accumulation of synonymous substitutions and a progressive increase of nonsynonymous substitutions over time, suggesting diversification likely was driven by positive selection. Short-term accumulation of nonsynonymous substitutions driven by selection may have significant implications for virulence, transmission dynamics, and even vaccine efficacy. Cholera, a dehydrating diarrheal disease caused by toxigenic strains of the bacterium Vibrio cholerae, emerged in 2010 in Haiti, a country where there were no available records on cholera over the past 100 years. While devastating in terms of morbidity and mortality, the outbreak provided a unique opportunity to study the evolutionary dynamics of V. cholerae and its environmental presence. The present study expands on previous work and provides an in-depth phylodynamic analysis inferred from genome-wide single nucleotide polymorphisms of clinical and environmental strains from dispersed geographic settings in

  1. Differential Expression Levels of Integrin α6 Enable the Selective Identification and Isolation of Atrial and Ventricular Cardiomyocytes.

    Directory of Open Access Journals (Sweden)

    Anne Maria Wiencierz

    Full Text Available Central questions such as cardiomyocyte subtype emergence during cardiogenesis or the availability of cardiomyocyte subtypes for cell replacement therapy require selective identification and purification of atrial and ventricular cardiomyocytes. However, current methodologies do not allow for a transgene-free selective isolation of atrial or ventricular cardiomyocytes due to the lack of subtype specific cell surface markers.In order to develop cell surface marker-based isolation procedures for cardiomyocyte subtypes, we performed an antibody-based screening on embryonic mouse hearts. Our data indicate that atrial and ventricular cardiomyocytes are characterized by differential expression of integrin α6 (ITGA6 throughout development and in the adult heart. We discovered that the expression level of this surface marker correlates with the intracellular subtype-specific expression of MLC-2a and MLC-2v on the single cell level and thereby enables the discrimination of cardiomyocyte subtypes by flow cytometry. Based on the differential expression of ITGA6 in atria and ventricles during cardiogenesis, we developed purification protocols for atrial and ventricular cardiomyocytes from mouse hearts. Atrial and ventricular identities of sorted cells were confirmed by expression profiling and patch clamp analysis.Here, we introduce a non-genetic, antibody-based approach to specifically isolate highly pure and viable atrial and ventricular cardiomyocytes from mouse hearts of various developmental stages. This will facilitate in-depth characterization of the individual cellular subsets and support translational research applications.

  2. Large-insert BAC/YAC libraries for selective re-isolation of genomic regions by homologous recombination in yeast.

    Science.gov (United States)

    Zeng, C; Kouprina, N; Zhu, B; Cairo, A; Hoek, M; Cross, G; Osoegawa, K; Larionov, V; de Jong, P

    2001-09-01

    We constructed representative large-insert bacterial artificial chromosome (BAC) libraries of two human pathogens (Trypanosoma brucei and Giardia lamblia) using a new hybrid vector, pTARBAC1, containing a yeast artificial chromosome (YAC) cassette (a yeast selectable marker and a centromere). The cassette allows transferring of BACs into yeast for their further modification. Furthermore, the new hybrid vector provides the opportunity to re-isolate each DNA insert without construction of a new library of random clones. Digestion of a BAC DNA by an endonuclease that has no recognition site in the vector, but which deletes most of the internal insert sequence and leaves the unique flanking sequences, converts a BAC into a TAR vector, thus allowing direct gene isolation. Cotransformation of a TAR vector and genomic DNA into yeast spheroplasts, and subsequent recombination between the TAR vector's flanking ends and a specific genomic fragment, allows rescue of the fragment as a circular YAC/BAC molecule. Here we prove a new cloning strategy by re-isolation of randomly chosen genomic fragments of different size from T. brucei cloned in BACs. We conclude that genomic regions of unicellular eukaryotes can be easily re-isolated using this technique, which provides an opportunity to study evolution of these genomes and the role of genome instability in pathogenicity.

  3. Isolation of Inositol Hexaphosphate (IHP)-Degrading Bacteria from Arbuscular Mycorrhizal Fungal Hyphal Compartments Using a Modified Baiting Method Involving Alginate Beads Containing IHP

    Science.gov (United States)

    Hara, Shintaro; Saito, Masanori

    2016-01-01

    Phytate (inositol hexaphosphate; IHP)-degrading microbes have been suggested to contribute to arbuscular mycorrhizal fungi (AMF)-mediated P transfer from IHP to plants; however, no IHP degrader involved in AMF-mediated P transfer has been isolated to date. We herein report the isolation of IHP-degrading bacteria using a modified baiting method. We applied alginate beads as carriers of IHP powder, and used them as recoverable IHP in the AM fungal compartment of plant cultivation experiments. P transfer from IHP in alginate beads via AMF was confirmed, and extracted DNA from alginate beads was analyzed by denaturing gradient gel electrophoresis targeting the 16S rRNA gene and a clone library method for the beta-propeller phytase (BPP) gene. The diversities of the 16S rRNA and BPP genes of microbes growing on IHP beads were simple and those of Sphingomonas spp. and Caulobacter spp. dominated. A total of 187 IHP-utilizing bacteria were isolated and identified, and they were consistent with the results of DNA analysis. Furthermore, some isolated Sphingomonas spp. and Caulobacter sp. showed IHP-degrading activity. Therefore, we successfully isolated dominant IHP-degrading bacteria from IHP in an AMF hyphal compartment. These strains may contribute to P transfer from IHP via AMF. PMID:27383681

  4. Selectable marker elimination in the T0 generation by Agrobacterium-mediated co-transformation involving Mungbean yellow mosaic virus TrAP as a non-conditional negative selectable marker and bar for transient positive selection.

    Science.gov (United States)

    RamanaRao, Mangu Venkata; Veluthambi, Karuppannan

    2010-05-01

    Transient selection involving the bar gene and non-conditional negative selection against stable T-DNA integration through the use of the Mungbean yellow mosaic virus (MYMV) transcriptional activator protein gene (TrAP) were used in a novel co-transformation strategy to generate selectable marker gene (SMG)-eliminated transgenic tobacco plants in the T(0) generation itself. Two compatible binary plasmids, pCam-bar-TrAP-gus harbouring bar as an SMG and the MYMV TrAP gene as a non-conditional negative selectable marker, and pGA472 with the nptII gene as an unselected experimental gene of interest (GOI) were placed in the Agrobacterium tumefaciens strain EHA105 and used for co-transformation. Transient selection with 5 mg l(-1) phosphinothricin (PPT) for 2-4 weeks and subsequent establishment in a PPT-minus medium yielded 114 plants from 200 leaf discs. The unselected nptII gene was detected by Southern blot analysis in 13 plants, revealing a co-transformation efficiency of 11.5%. Five of these plants harboured only the nptII gene (GOI) and not the bar gene (SMG). Thus, SMG elimination was achieved in the T(0) generation itself in 4.4% (5/114) of plants, which were transiently selected for 2-4 weeks on PPT. MYMV TrAP, a non-conditional negative selectable marker, effectively reduced the recovery of plants with stable integration of the SMG (bar).

  5. Neovestitol, an isoflavonoid isolated from Brazilian red propolis, reduces acute and chronic inflammation: involvement of nitric oxide and IL-6

    OpenAIRE

    Marcelo Franchin; Colón, David F.; da Cunha, Marcos G; Castanheira, Fernanda V. S.; André L. L. Saraiva; Bruno Bueno-Silva; Alencar,Severino M.; Cunha, Thiago M; Rosalen, Pedro L.

    2016-01-01

    Isoflavonoids have been largely studied due to their distinct biological activities identified thus far. Herein, we evaluated the activity of neovestitol, an isoflavonoid isolated from Brazilian red propolis, in acute and chronic inflammation. As for acute inflammation, we found that neovestitol reduced neutrophil migration, leukocyte rolling and adhesion, as well as expression of ICAM-1 in the mesenteric microcirculation during lipopolysaccharide-induced acute peritonitis. No changes were ob...

  6. Method of selecting vibro-isolation properties of vibration reduction systems

    Energy Technology Data Exchange (ETDEWEB)

    Maciejewski, Igor; Krzyzynski, Tomasz [Koszalin University of Technology, Koszalin (Poland)

    2016-04-15

    An effective optimization method is proposed in this paper to determine the basic characteristics of non-linear visco-elastic elements used in passive vibration reduction systems. The developed method is examined by performing experimental investigations on the exemplary vibration reduction system, i.e., the passive seat suspension. The shaping of vibro-isolation properties is analyzed for different spectral classes of excitation signals.

  7. Core analyses for selected samples from the Culebra Dolomite at the Waste Isolation Pilot Plant site

    Energy Technology Data Exchange (ETDEWEB)

    Kelley, V.A.; Saulnier, G.J. Jr. (INTERA, Inc., Austin, TX (USA))

    1990-11-01

    Two groups of core samples from the Culebra Dolomite Member of the Rustler Formation at and near the Waste Isolation Pilot Plant were analyzed to provide estimates of hydrologic parameters for use in flow-and-transport modeling. Whole-core and core-plug samples were analyzed by helium porosimetry, resaturation and porosimetry, mercury-intrusion porosimetry, electrical-resistivity techniques, and gas-permeability methods. 33 refs., 25 figs., 10 tabs.

  8. Core analyses for selected samples from the Culebra Dolomite at the Waste Isolation Pilot Plant site

    Energy Technology Data Exchange (ETDEWEB)

    Kelley, V.A.; Saulnier, G.J. Jr. (INTERA, Inc., Austin, TX (USA))

    1990-11-01

    Two groups of core samples from the Culebra Dolomite Member of the Rustler Formation at and near the Waste Isolation Pilot Plant were analyzed to provide estimates of hydrologic parameters for use in flow-and-transport modeling. Whole-core and core-plug samples were analyzed by helium porosimetry, resaturation and porosimetry, mercury-intrusion porosimetry, electrical-resistivity techniques, and gas-permeability methods. 33 refs., 25 figs., 10 tabs.

  9. Isolating "Unknown" Bacteria in the Introductory Microbiology Laboratory: A New Selective Medium for Gram-Positives.

    Science.gov (United States)

    McKillip, John L.; Drake, MaryAnne

    1999-01-01

    Describes the development, preparation, and use of a medium that can select against a wide variety of Gram-negative bacteria while still allowing growth and differentiation of a wide range of Gram-positives. (WRM)

  10. Two new opioid delta-receptor ligands: a highly selective agonist and a potent selective antagonist in in vitro isolated preparations.

    Science.gov (United States)

    Ueki, M; Aoki, K; Kajiwara, M; Shinozaki, K; Inoue, H; Oka, T

    1984-12-01

    N,N-Diallyl derivatives of enkephalin analogues were chemically synthesized, and their biological activities were estimated in vitro isolated preparations. N,N-Diallyl-[D-Ala2, D-Leu5]-enkephalin [test compound I] at doses up to 10 microM did not inhibit the electrically-evoked contractions of guinea-pig ileum, which had been suggested to contain opioid mu- and kappa-receptors, but it significantly depressed the contractions of mouse vas deferens, which had been indicated to contain mu-, kappa- and delta-receptors, suggesting that test compound I did not act on both mu- and kappa-receptors, but acted on delta-receptors. Additionally, the Ke (equilibrium dissociation constant) values against test compound I of naloxone were approximately 30 nM and similar to those of Mr 2266, also indicating that test compound I acted as a delta agonist. Moreover, the Ke values of ICI 154129 against compound I were approximately 340 nM, strongly suggesting that test compound I acted as a delta agonist. The Ke values of bis-[N,N-diallyl-[D-Ala2, Leu5]-enkephalyl]-cystine [test compound II] against [D-Ala2, D-Leu5]-enkephalin in mouse vas deferens and morphine or ethylketocyclazocine in guinea-pig ileum were 44.9 nM and 5.00 or 11.3 microM, respectively, showing that test compound II was a potent selective opioid delta antagonist. In conclusion, among compounds synthesized, two new opioid delta-receptor ligands, one being a highly selective agonist and the other being a potent selective antagonist in in vitro isolated preparations, were found in the present study.

  11. Selected lactic acid-producing bacterial isolates with the capacity to reduce Salmonella translocation and virulence gene expression in chickens.

    Science.gov (United States)

    Yang, Xiaojian; Brisbin, Jennifer; Yu, Hai; Wang, Qi; Yin, Fugui; Zhang, Yonggang; Sabour, Parviz; Sharif, Shayan; Gong, Joshua

    2014-01-01

    Probiotics have been used to control Salmonella colonization/infection in chickens. Yet the mechanisms of probiotic effects are not fully understood. This study has characterized our previously-selected lactic acid-producing bacterial (LAB) isolates for controlling Salmonella infection in chickens, particularly the mechanism underlying the control. In vitro studies were conducted to characterize 14 LAB isolates for their tolerance to low pH (2.0) and high bile salt (0.3-1.5%) and susceptibility to antibiotics. Three chicken infection trials were subsequently carried out to evaluate four of the isolates for reducing the burden of Salmonella enterica serovar Typhimurium in the broiler cecum. Chicks were gavaged with LAB cultures (10(6-7) CFU/chick) or phosphate-buffered saline (PBS) at 1 day of age followed by Salmonella challenge (10(4) CFU/chick) next day. Samples of cecal digesta, spleen, and liver were examined for Salmonella counts on days 1, 3, or 4 post-challenge. Salmonella in the cecum from Trial 3 was also assessed for the expression of ten virulence genes located in its pathogenicity island-1 (SPI-1). These genes play a role in Salmonella intestinal invasion. Tested LAB isolates (individuals or mixed cultures) were unable to lower Salmonella burden in the chicken cecum, but able to attenuate Salmonella infection in the spleen and liver. The LAB treatments also reduced almost all SPI-1 virulence gene expression (9 out of 10) in the chicken cecum, particularly at the low dose. In vitro treatment with the extracellular culture fluid from a LAB culture also down-regulated most SPI-1 virulence gene expression. The possible correlation between attenuation of Salmonella infection in the chicken spleen and liver and reduction of Salmonella SPI-1 virulence gene expression in the chicken cecum by LAB isolates is a new observation. Suppression of Salmonella virulence gene expression in vivo can be one of the strategies for controlling Salmonella infection in chickens.

  12. Identification of Highly Potent and Selective α-Glucosidase Inhibitors with Antiglycation Potential, Isolated from Rhododendron arboreum

    OpenAIRE

    Rabia Raza; Zaitoon Ilyas; Sajid Ali; Muhammad Nisar; Muhammad Younas Khokhar; Jamshed Iqbal

    2015-01-01

    This study explored antidiabetic potential of eight known pure compounds, isolated from the bark of Rhododendron arboreum. Invitro studies of these compounds against α and β-glucosidases revealed them as very potent and selective inhibitors of α-glucosidase. Compound 7 (3-O-acetylursolic acid) was found to be the most potent inhibitor of α-glucosidase with 3.3±0.1µM IC 50 value which was many folds higher than standard inhibitor acarbose. Antiglycation studies of compounds showed that all com...

  13. Potency and selectivity indices of acetone leaf extracts of nine selected South African trees against six opportunistic Enterobacteriaceae isolates from commercial chicken eggs.

    Science.gov (United States)

    Elisha, Ishaku L; Jambalang, Alexander R; Botha, Francien S; Buys, Elna M; McGaw, Lyndy J; Eloff, Jacobus N

    2017-02-02

    The rise in antimicrobial resistance in a plethora of nosocomial and opportunistic bacterial pathogens often isolated from commercial eggs, poses a serious public health concern. The existence of these contaminants may also serve as a drawback in the current efforts of improving the well-being of immunocompromised patients. The aim of this study was to determine the efficacy of plant extracts that had good activity on Escherichia coli in previous word on pathogens isolated from eggs for possible use in combating pathogens from eggs. Acetone leaf extracts of nine trees with good activities against Escherichia coli were tested for their in vitro antibacterial activity against six opportunistic bacterial isolates from commercial eggs (Stenotrophomonas maltophilia, Klebsiella pneumoniae, Salmonella serotype Typhimurium, Proteus mirabilis, Enterobacter cloacae and Escherichia coli) using a serial microdilution method with tetrazolium violet as indicator of growth. Cytotoxicity was determined using a tetrazolium-based colorimetric assay against Vero kidney cells, and selectivity index calculated. The MIC values range of the different extracts against Stenotrophomonas maltophilia was 0.08-0.31 mg/ml, Klebsiella pneumonia 0.08-0.63 mg/ml, Salmonella ser. Typhimurium 0.08-0.63 mg/ml, Proteus mirabilis 0.02-1.25 mg/ml, Enterobacter cloacae 0.08-0.31 mg/ml and Escherichia coli 0.08-0.16 mg/ml respectively. Escherichia coli was the most sensitive while Proteus mirabilis was most resistant pathogen to the different test extracts, with mean MIC values of 0.08 mg/ml and 0.46 mg/ml respectively. Cremaspora triflora extracts had good activity against all the pathogenic egg isolates, with the exception of Proteus mirabilis. Maesa lanceolata and Elaeodendron croceum had the best total antibacterial activity (TAA), while generally the selectivity index of the extract was low (SI activity of C. triflora extracts suggests that the plant has potential as a therapeutic agent

  14. Isolation of keratinophilic fungi from selected soils of Sanjay Gandhi National Park, Mumbai (India).

    Science.gov (United States)

    Deshmukh, S K; Verekar, S A

    2014-12-01

    One hundred and twenty-five samples were collected from eight different sites in the vicinity of Sanjay Gandhi National Park (SGNP) and screened for the presence of keratinophilic fungi using hair baiting technique for isolation. Seventy-three isolates were recovered and identified. The cultures were identified using macro- and micro-morphological features. Their identification was also confirmed by the BLAST search of sequences of the ITS1-5.8S-ITS2 rDNA region against the NCBI/Genbank data and compared with deposited sequences for identification purpose. Thirteen species of nine genera were isolated viz. Aphanoascus durus (2.4%), Arthroderma corniculatum (1.6%), Auxarthron umbrinum (0.8%), Chrysosporium evolceanui (1.6%), Chrysosporium indicum (16.0%), Chrysosporium tropicum (2.4%), Chrysosporium zonatum (4.0%), Chrysosporium states of Arthroderma tuberculatum (0.8%), Chrysosporium state of Ctenomyces serratus (11.2%), Gymnascella dankaliensis (3.2%), Microsporum gypseum (12.0%), Myriodontium keratinophilum (0.8%) and Trichophyton mentagrophytes (1.6%). Representative of all thirteen species can release the protein in the range of 152.2-322.4 μg/mL in liquid media when grown on human hair in shake flask culture and also decompose 18.4-40.2% of human hair after four weeks of incubation. This study indicates that the soils of SGNP, Mumbai may be significant reservoirs of certain keratinophilic fungi. The keratinolytic activity of these fungi may be playing significant role in superficial infections to man and animals and recycling of keratinic material of this environment.

  15. Enhancement of Magneto-Optic Effect in Optical Isolator with Semiconductor Guiding Layer by Selective Oxidation of AlInAs

    Institute of Scientific and Technical Information of China (English)

    Takashi; Sakai; Yuya; Shoji; Hideki; Yokoi; Tetsuya; Mizumoto

    2003-01-01

    Selective oxidation of an AlInAs layer was investigated for enhancement of magneto-optic effect in an optical isolator. Twelve times nonreciprocal phase shift enhancement was estimated from a measured AlInAs-oxide refractive index.

  16. Steroid-responsive IgG4-related disease with isolated prostatic involvement: An unusual presentation with elevated serum PSA

    Directory of Open Access Journals (Sweden)

    Vikas Jain

    2016-01-01

    Full Text Available Autoimmune prostatitis is known to occur as a part of multisystem fibro-inflammatory disorder known as IgG4 related disease (IgG4 RD. The usual presentation is with symptoms of gastro-intestinal disease with prostatic involvement presenting as lower urinary tract symptoms. The disease responds to corticosteroids. We report an asymptomatic young man who was diagnosed to have IgG4 related prostatitis on TRUS-guided prostate biopsy done for elevated serum PSA, in the absence of any other systemic involvement. The treatment with steroid resulted in normalization of S PSA levels.

  17. Selection of lactic acid bacteria isolated from Tunisian cereals and exploitation of the use as starters for sourdough fermentation.

    Science.gov (United States)

    Mamhoud, Asma; Nionelli, Luana; Bouzaine, Taroub; Hamdi, Moktar; Gobbetti, Marco; Rizzello, Carlo Giuseppe

    2016-05-16

    Wheat bread is the most popular staple food consumed in Tunisia and, despite the niche production of some typical breads (e.g. Tabouna, Mlawi, Mtabga), the major part is currently produced with baker's yeast at industrial or, mainly, at artisanal level, while the use of sourdough fermentation is rarely reported. Considering the growing national demand for cereal baked goods, it can be hypothesized that sourdough fermentation through the use of selected lactic acid bacteria as starters could improve the overall quality and the diversification of local products. Different cereal grains were collected from the regions of Ariana, Bizerta, Beja Nabeul, and Seliana, and the autochthonous lactic acid bacteria were isolated, identified, characterized and selected on the basis of the kinetics of acidification, the proteolytic activity, and the quotient of fermentation. Lactobacillus curvatus MA2, Pediococcus pentosaceus OA2, and Pediococcus acidilactici O1A1 were used together as mixed starter to obtain a selected sourdough. According to the backslopping procedure, a type I sourdough was made from a Tunisian flour (spontaneous sourdough). Compared to the use of the spontaneous sourdough, the one obtained with selected and mixed starters by a unique fermentation step, favored the increase of the concentrations of organic acids, phenols, and total free amino acids, the most suitable quotient of fermentation, and the most intense phytase and antioxidant activities, that increased ca. 20% compared to the control. Moreover, the selected starters improved the in vitro protein digestibility (ca. 82% when selected sourdough was used), textural and sensory features of the breads, as determined by textural profile analysis and panel test, respectively. This study aimed at exploiting the potential of selected autochthonous lactic acid bacteria and extending the use of a sourdough (type II), thanks to the set-up of a two-step fermentation protocol designed for application at the

  18. Antihepatic Fibrosis Effect of Active Components Isolated from Green Asparagus (Asparagus officinalis L.) Involves the Inactivation of Hepatic Stellate Cells.

    Science.gov (United States)

    Zhong, Chunge; Jiang, Chunyu; Xia, Xichun; Mu, Teng; Wei, Lige; Lou, Yuntian; Zhang, Xiaoshu; Zhao, Yuqing; Bi, Xiuli

    2015-07-08

    Green asparagus (Asparagus officinalis L.) is a vegetable with numerous nutritional properties. In the current study, a total of 23 compounds were isolated from green asparagus, and 9 of these compounds were obtained from this genus for the first time. Preliminary data showed that the ethyl acetate (EtOAc)-extracted fraction of green asparagus exerted a stronger inhibitory effect on the growth of t-HSC/Cl-6 cells, giving an IC50 value of 45.52 μg/mL. The biological activities of the different compounds isolated from the EtOAc-extracted fraction with respect to antihepatic fibrosis were investigated further. Four compounds, C3, C4, C10, and C12, exhibited profound inhibitory effect on the activation of t-HSC/Cl-6 cells induced by TNF-α. The activation t-HSC/Cl-6 cells, which led to the production of fibrotic matrix (TGF-β1, activin C) and accumulation of TNF-α, was dramatically decreased by these compounds. The mechanisms by which these compounds inhibited the activation of hepatic stellate cells appeared to be associated with the inactivation of TGF-β1/Smad signaling and c-Jun N-terminal kinases, as well as the ERK phosphorylation cascade.

  19. Selective isolation of β-glucan from corn pericarp hemicelluloses by affinity chromatography on cellulose column.

    Science.gov (United States)

    Yoshida, Tomoki; Honda, Yoichi; Tsujimoto, Takashi; Uyama, Hiroshi; Azuma, Jun-ichi

    2014-10-13

    A combination of anion-exchange chromatography and affinity chromatography on a cellulose column was found to be effective for the isolation of β-(1,3;1,4)-glucan (BG) from corn pericarp hemicelluloses (CPHs). CPHs containing 6.6% BG were extracted from corn pericarp with 6M urea-2 wt% NaOH solution and initially fractionated into neutral and acidic parts by anion exchange chromatography to remove acidic arabinoxylan consisting of arabinose (35.6%) and xylose (50.9%). The neutral fraction (yield; 10.1% on the basis of CPHs) consisting of 1.0% arabinose, 10.1% xylose and 80.3% glucose containing 28.4% BG was then applied to a cellulose column of Whatman CF-11. BG could be recovered from the adsorbed fraction on the cellulose column by elution with 2% NaOH in a yield of 2.6% on the basis of CPHs with a purity of 84.7%. The chemical structure of the isolated corn pericarp BG was confirmed by (13)C NMR spectroscopic, methylation and lichenase treatment analyses. The results indicate that the ratios of (1,4)/(1,3) linkage and cellotriosyl/cellotetraosyl segments of the BG were 2.60 and 2.5, respectively.

  20. Cytotoxicity of syringin and 4-methoxycinnamyl alcohol isolated from Foeniculum vulgare on selected human cell lines.

    Science.gov (United States)

    Lall, Namrita; Kishore, Navneet; Binneman, Brigitte; Twilley, Danielle; van de Venter, Maryna; du Plessis-Stoman, Debbie; Boukes, Gerhardt; Hussein, Ahmed

    2015-01-01

    This study was carried out to determine the cytotoxic effect of seven plant extracts and the isolated compounds - syringin and 4-methoxycinnamyl alcohol - on cancerous and non-cancerous cells. The ethanol extract of Foeniculum vulgare was found to exhibit the most significant toxicity with an IC50 value of 19.97 μg/mL on HeLa cells. Bioassay-guided fractionation led to the isolation of two compounds, syringin (1) and 4-methoxycinnamyl alcohol (2). Both compounds showed toxicity against MCF-7, HeLa and DU145 cancer cell line. The results showed that compound 2 showed high toxicity against all the cancer cell lines with IC50 values of 14.24, 7.82 and 22.10 μg/mL, respectively. 4-Methoxycinnamyl alcohol also showed no apoptotic effect in cell cycle analysis after 48 h at a concentration of 10 μg/mL. However, DNA fragmentation study revealed that necrosis took place at a concentration of 10 μg/mL after 48 h exposure.

  1. Selection and characterisation of an HD1-like Bacillus thuringiensis isolate with a high insecticidal activity against Spodoptera littoralis (Lepidoptera: Noctuidae).

    Science.gov (United States)

    Azzouz, Hichem; Kebaili-Ghribi, Jihene; ben Farhat-Touzri, Dalel; Daoud, Fatma; Fakhfakh, Ines; Tounsi, Slim; Jaoua, Samir

    2014-08-01

    Spodoptera littoralis (Boisduval) larvae are known by their susceptibility to Bacillus thuringiensis subsp. aizawai strains. In order to prevent the appearance of B. thuringiensis (Bt) resistance and to develop economical Bt-based biopesticides, the selection and the characterisation of a B. thuringiensis isolate toxic against S. littoralis larvae and overproducing δ-endotoxins were investigated. Among 124 Tunisian B. thuringiensis isolates assessed against S. littoralis larvae, four isolates showed toxicity similar to and higher than the toxicity of the aizawai strain HD133 and the kurstaki strain HD1 respectively. The plasmid pattern of the selected isolates was similar to that of HD1. Polymerase chain reaction (PCR) analysis using specific primers revealed that these isolates present different gene contents. The only detected gene encoding Spodoptera-specific toxin was cry9. The selected isolates were found to produce bipyramidal and cubic crystals. The assessment of δ-endotoxin production by these isolates showed that BUPM28 produced 43.71 and 80.81% more δ-endotoxin than HD1 and HD133 respectively. The application of osmotic or heat shock stress on the BUPM28 isolate made it possible to enhance δ-endotoxin production by 22 and 23% respectively. On the basis of its potent insecticidal activity and its high level of δ-endotoxin production, the BUPM28 isolate can be considered to be an effective alternative for the control of S. littoralis. © 2013 Society of Chemical Industry.

  2. Isolation and characterization of Lotus japonicus genes involved in iron and zinc homeostasis

    DEFF Research Database (Denmark)

    Cvitanich, Cristina; Jensen, Winnie; Sandal, Niels Nørgaard;

    in plants. We have used these sequences to search for L. japonicus ESTs and genomic loci that are likely to be involved in iron and zinc metabolism. We have identified sequences corresponding to ferritins, ferric reductases, metal transport proteins of the ZIP family, and cation transporters of the NRAMP....... japonicus Gifu recombinant inbred lines....

  3. Selection of Trypanosoma cruzi clonal genotypes (clonet 20 and 39) isolated from Bolivian triatomines following subculture in liquid medium.

    Science.gov (United States)

    Bosseno, M F; Yacsik, N; Vargas, F; Brenière, S F

    2000-01-01

    Previous studies showed that two groups of Trypanosoma cruzi clonal genotypes named clonet 20 and clonet 39 were predominant in Triatoma infestans, the unique vector of Chagas disease in Bolivia. These groups of clones correspond to distinct genetic clusters. These clonets were detected in T. infestans and Rhodnius pictipes fecal samples before isolation and after culture by kDNA PCR (polymerase chain reaction) and hybridization of the amplified products with clonet specific kDNA probes named 20 and 39 as previously reported. Forty eight T. infestans and three R. pictipes infected insects captured at random in different Bolivian departments were proceeded. As previously reported the direct identification of the two major clonets in fecal samples allowed the detection of abundant mixed infections: 41% in the original sample, however after culture, only 6% of mixed infections were detected. Among the 21 parasite stocks isolated from digestive tracts where mixed infections were initially detected (clonet 20 + 39) clonet 20 alone was detected in 81% of them. This result clearly showed that the culture step selected clonet 20 parasites over those belonging to clonet 39. The taxonomic status of the isolated stocks was also confirmed by isoenzyme typing, and correlation was observed between clustering topology and hybridization patterns with the probes 20 and 39.

  4. In vitro screening of selected probiotic properties of Lactobacillus strains isolated from traditional fermented cabbage and cucumber.

    Science.gov (United States)

    Zielińska, Dorota; Rzepkowska, Anna; Radawska, Anna; Zieliński, Konrad

    2015-02-01

    Most important during probiotic selection are gastric acid and bile tolerance, the adhesion to the luminal epithelium to colonize the lower gastrointestinal tract of a human and safety for human consumption. The aim of this study was to evaluate the selected probiotic in vitro properties of Lactobacillus spp. Strains isolated from traditional fermented food. A total 38 strains were isolated from the pickled samples and 14 were identified as Lactobacillus spp. The survival of almost all strains after incubation at pH 2.5 did not change markedly, and remained at above 90 % (10(9) CFU/mL). The strains also exhibited a high survival rate at pH 3.5 (>90 %), whereas pH 1.5 all were died. Just four strains could survive 90 min. at pH 1.5 (Lactobacillus strains are safe and can survive under gastrointestinal conditions, which requires them to future in vitro and in vivo probiotic studies.

  5. Parental Involvement and Home Environment in Music: Current and Former Students from Selected Community Music Programs in Brazil and the United States

    Science.gov (United States)

    Barnes, Gail V.; DeFreitas, Aureo; Grego, John

    2016-01-01

    The purpose of this study was to determine whether individuals' perceptions of parental involvement and home environment in music vary with nationality (Brazil/United States) and time frame (past/current). Past and current students from selected community music programs in the United States and Brazil completed the PI-HEM (Parental Involvement and…

  6. Ureaplasma parvum undergoes selection in utero resulting in genetically diverse isolates colonizing the chorioamnion of fetal sheep.

    Science.gov (United States)

    Dando, Samantha J; Nitsos, Ilias; Polglase, Graeme R; Newnham, John P; Jobe, Alan H; Knox, Christine L

    2014-02-01

    Ureaplasmas are the microorganisms most frequently isolated from the amniotic fluid of pregnant women and can cause chronic intrauterine infections. These tiny bacteria are thought to undergo rapid evolution and exhibit a hypermutatable phenotype; however, little is known about how ureaplasmas respond to selective pressures in utero. Using an ovine model of chronic intraamniotic infection, we investigated if exposure of ureaplasmas to subinhibitory concentrations of erythromycin could induce phenotypic or genetic indicators of macrolide resistance. At 55 days gestation, 12 pregnant ewes received an intraamniotic injection of a nonclonal, clinical Ureaplasma parvum strain followed by (i) erythromycin treatment (intramuscularly, 30 mg/kg/day, n = 6) or (ii) saline (intramuscularly, n = 6) at 100 days gestation. Fetuses were then delivered surgically at 125 days gestation. Despite injecting the same inoculum into all the ewes, significant differences between amniotic fluid and chorioamnion ureaplasmas were detected following chronic intraamniotic infection. Numerous polymorphisms were observed in domain V of the 23S rRNA gene of ureaplasmas isolated from the chorioamnion (but not the amniotic fluid), resulting in a mosaiclike sequence. Chorioamnion isolates also harbored the macrolide resistance genes erm(B) and msr(D) and were associated with variable roxithromycin minimum inhibitory concentrations. Remarkably, this variability occurred independently of exposure of ureaplasmas to erythromycin, suggesting that low-level erythromycin exposure does not induce ureaplasmal macrolide resistance in utero. Rather, the significant differences observed between amniotic fluid and chorioamnion ureaplasmas suggest that different anatomical sites may select for ureaplasma subtypes within nonclonal, clinical strains. This may have implications for the treatment of intrauterine ureaplasma infections.

  7. Surface assembly of graphene oxide nanosheets on SiO2 particles for the selective isolation of hemoglobin.

    Science.gov (United States)

    Liu, Jia-Wei; Zhang, Qian; Chen, Xu-Wei; Wang, Jian-Hua

    2011-04-18

    Graphene oxide (GO) nanosheets have been immobilized onto SiO(2) particles through electrostatic interactions by surface assembly. The surface-assembled composite material was characterized by means of SEM and FTIR and UV/Vis spectroscopy to reveal an assembling ratio of 2.3% (w/w, GO/SiO(2)). The GO/SiO(2) composites were subsequently used, for the first time, as adsorbents for the adsorption/isolation of proteins. Selective isolation of proteins of interest, namely, hemoglobin (Hb) in this case, from complex sample matrices, for example, human whole blood, could be obtained by carefully manipulating the adsorption/desorption process. At pH 7, an adsorption of 85% was achieved for Hb (70 mg L(-1)) in sample solution (1.0 mL) by the GO/SiO(2) composites (3.0 mg). The adsorption behavior was consistent with the Langmuir adsorption model, corresponding to a theoretical adsorption capacity of 50.5 mg g(-1) for Hb. The retained Hb could be readily recovered by using a Tris-HCl buffer at pH 8.9 to give a recovery of 80%. Circular dichroism and specific activity investigations indicated that the GO/SiO(2) composites exhibited favorable biocompatibility, characterized by virtually no effect on the conformation and activity of Hb after adsorption/desorption. The composites were used for the selective isolation of Hb from human whole blood and achieved satisfactory results by assaying with sodium dodecyl sulfate polyacrylamide gel electrophoresis.

  8. Isolation and characterization of a novel, highly selective astaxanthin-producing marine bacterium.

    Science.gov (United States)

    Asker, Dalal

    2017-09-18

    A high throughput screening approach for astaxanthin-producing bacteria led to the discovery of a novel highly selective astaxanthin-producing marine bacterium (strain N-5). Phylogenetic analysis based on partial 16S rRNA gene and phenotypic metabolic testing indicated it belongs to the genus Brevundimonas. Therefore, it was designated as Brevundimonas sp. strain N-5. To identify and quantify carotenoids produced by strain N-5, HPLC-DAD and HPLC-MS methods were used. The culture conditions including media, shaking and time had significant effects on cell growth and carotenoids production including astaxanthin. The total carotenoids were ~601.2 µg g-1 dry cells including a remarkable amount (364.6 µg g-1 dry cells) of optically pure astaxanthin (3S, 3'S) isomer, with high selectivity (~60.6%) under medium aeration conditions. Notably, increasing the culture aeration enhanced astaxanthin production up to 85% of total carotenoids. This is the first report that describes a natural, highly selective astaxanthin-producing marine bacterium.

  9. Glyceollins and dehydroglyceollins isolated from soybean act as SERMs and ER subtype-selective phytoestrogens.

    Science.gov (United States)

    van de Schans, Milou G M; Vincken, Jean-Paul; de Waard, Pieter; Hamers, Astrid R M; Bovee, Toine F H; Gruppen, Harry

    2016-02-01

    Seven prenylated 6a-hydroxy-pterocapans and five prenylated 6a,11a-pterocarpenes with different kinds of prenylation were purified from an ethanolic extract of fungus-treated soybean sprouts. The activity of these compounds toward both human estrogen receptors (hERα and hERβ) was determined in a yeast bioassay and the activity toward hERα was additionally tested in an U2-OS based hERα CALUX bioassay. In the yeast bioassay, compounds with chain prenylation showed in general an agonistic mode of action toward hERα, whereas furan and pyran prenylation led to an antagonistic mode of action. Five of these antagonistic compounds had an agonistic mode of action in the U2-OS based hERα CALUX bioassay, implying that these compounds can act as SERMs. The yeast bioassay also identified 8 ER subtype-selective compounds, with either an antagonistic mode of action or no response toward hERα and an agonistic mode of action toward hERβ. The ER subtype-selective compounds were characterized by 6a-hydroxy-pterocarpan or 6a,11a-pterocarpene backbone structure. It is suggested that either the extra D-ring or the increase in length to 12-13.5Å of these compounds is responsible for an agonistic mode of action toward hERβ and, thereby, inducing ER subtype-selective behavior.

  10. Nuclear substructure reorganization during late stageerythropoiesis is selective and does not involve caspase cleavage ofmajor nuclear substructural proteins

    Energy Technology Data Exchange (ETDEWEB)

    Krauss, Sharon Wald; Lo, Annie J.; Short, Sarah A.; Koury, MarkJ.; Mohandas, Narla; Chasis, Joel Anne

    2005-04-06

    Enucleation, a rare feature of mammalian differentiation, occurs in three cell types: erythroblasts, lens epithelium and keratinocytes. Previous investigations suggest that caspase activation functions in lens epithelial and keratinocyte enucleation, as well as in early erythropoiesis encompassing BFU-E differentiation to proerythroblast. To determine whether caspase activation contributes to later erythropoiesis and whether nuclear substructures other than chromatin reorganize, we analyzed distributions of nuclear subcompartment proteins and assayed for caspase-induced cleavage of subcompartmental target proteins in mouse erythroblasts. We found that patterns of lamin B in the filamentous network interacting with both the nuclear envelope and DNA, nuclear matrix protein NuMA, and splicing factors Sm and SC35 persisted during nuclear condensation, consistent with effective transcription of genes expressed late in differentiation. Thus nuclear reorganization prior to enucleation is selective, allowing maintenance of critical transcriptional processes independent of extensive chromosomal reorganization. Consistent with these data, we found no evidence for caspase-induced cleavage of major nuclear subcompartment proteins during late erythropoiesis, in contrast to what has been observed in early erythropoiesis and in lens epithelial and keratinocyte differentiation. These findings imply that nuclear condensation and extrusion during terminal erythroid differentiation involve novel mechanisms that do not entail major activation of apoptotic machinery.

  11. Selection during crop diversification involves correlated evolution of the circadian clock and ecophysiological traits in Brassica rapa.

    Science.gov (United States)

    Yarkhunova, Yulia; Edwards, Christine E; Ewers, Brent E; Baker, Robert L; Aston, Timothy Llewellyn; McClung, C Robertson; Lou, Ping; Weinig, Cynthia

    2016-04-01

    Crop selection often leads to dramatic morphological diversification, in which allocation to the harvestable component increases. Shifts in allocation are predicted to impact (as well as rely on) physiological traits; yet, little is known about the evolution of gas exchange and related anatomical features during crop diversification. In Brassica rapa, we tested for physiological differentiation among three crop morphotypes (leaf, turnip, and oilseed) and for correlated evolution of circadian, gas exchange, and phenological traits. We also examined internal and surficial leaf anatomical features and biochemical limits to photosynthesis. Crop types differed in gas exchange; oilseed varieties had higher net carbon assimilation and stomatal conductance relative to vegetable types. Phylogenetically independent contrasts indicated correlated evolution between circadian traits and both gas exchange and biomass accumulation; shifts to shorter circadian period (closer to 24 h) between phylogenetic nodes are associated with higher stomatal conductance, lower photosynthetic rate (when CO2 supply is factored out), and lower biomass accumulation. Crop type differences in gas exchange are also associated with stomatal density, epidermal thickness, numbers of palisade layers, and biochemical limits to photosynthesis. Brassica crop diversification involves correlated evolution of circadian and physiological traits, which is potentially relevant to understanding mechanistic targets for crop improvement.

  12. SELECTION OF BEAUVERIA BASSIANA (BALSAMO VUILLEMIN ISOLATES FOR MANAGEMENT OF MYZUS PERSICAE (SULTZAR (HOM.: APHIDAE BASED ON VIRULENCE AND GROWTH RELATED CHARACTERISTICS

    Directory of Open Access Journals (Sweden)

    Mohammad S. Al-alawi

    2014-01-01

    Full Text Available Isolates of the entomopathogenic fungus Beauveria bassiana originated from Jordan were evaluated for their efficacy against the green peach aphid, Myzus persicae under laboratory and greenhouse conditions. Efficacy evaluation involved testing all isolates at a concentration of 1×107 conidia/mL followed by concentration dependent and greenhouse bioassays for the top virulent isolates. Growth characteristics related to virulence were evaluated for high, intermediate and low virulent isolates. Results showed that three isolates namely: BAU004, BAU018 and BAU019 were highly virulent to the aphid in the laboratory causing more than 75% infection. In the greenhouse, the three isolates caused infection from 41.3 to 46.5%. For the growth characteristics, isolate BAU019 produced more spores than the other highly virulent ones including the commercial isolate GHA. Highly virulent isolates also showed faster hyphal growth than low virulent isolates. These findings indicate that isolates BAU004, Bau018 and BAU019 might be developed as commercial microbial insecticides for safe and effective control of green peach aphid.

  13. Isolation of a Tomato Protease that May Be Involved in Proteolysis of 1-Aminocyclopropane-1-Carboxylate Synthase

    Institute of Scientific and Technical Information of China (English)

    Jian-Feng LI; Liang-Hu QU; Ning LI

    2005-01-01

    1-aminocyclopropane-1-carboxylate (ACC) synthase is a principal enzyme that catalyses the committed step in phytohormone ethylene biosynthesis. Previous evidence indicates that the hypervariable C-terminus of ACC synthase is most likely to be processed proteolytically in vivo. However, the protease responsible has not been identified thus far. In the present study, we detected proteolytic activity against ACC synthase (LeACS2) in tomato (Lycopersicon esculentum Mill.) fruit extract based on a newly established in vitro assay system. Purification of the protease through DEAE, gel filtration and MonoQ chromatography resulted in considerable enrichment of a 64-kDa protein species. Subsequent biochemical analysis of the purified tomato protease revealed that the optimal conditions for its proteolytic activity were at pH 8.0 and at 37 ℃. In addition, the protease activity was blocked completely by the metalloprotease inhibitor 1,10-phenanthroline. The present study represents the first report on the isolation of an ACC synthaseprocessing protease from plant tissues.

  14. The involvement of cytochrome P450 peroxidase in the metabolic bioactivation of cumene hydroperoxide by isolated rat hepatocytes.

    Science.gov (United States)

    Anari, M R; Khan, S; O'Brien, P J

    1996-09-01

    Organic hydroperoxides are believed to be primarily detoxified in cells by the GSH peroxidase/GSSG reductase system and activated to cytotoxic radical species by non-heme iron. However, organic hydroperoxides seem to be bioactivated by cytochrome P450 (P450) in isolated hepatocytes as various P450 (particularly P450 2E1) inhibitors inhibited cumene hydroperoxide (CumOOH) metabolism and attenuated subsequent cytotoxic effects including antimycin A-resistant respiration, lipid peroxidation, iron mobilization, ATP depletion, and cell membrane disruption. CumOOH metabolism was also faster in P450 1A-induced hepatocytes and was inhibited by the P450 1A inhibitor alpha-naphthoflavone. The ferric chelator deferoxamine also prevented cytotoxicity even after CumOOH had been metabolized but had no effect on CumOOH metabolism. This emphasizes the toxicological significance of the iron released following hydroperoxide metabolic activation by cytochrome P450. The radical trap, 4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPO), had no effect on CumOOH metabolism but prevented CumOOH-induced antimycin A-resistant respiration, lipid peroxidation, iron mobilization, and loss of membrane integrity. These results suggest that CumOOH is metabolically activated by some P450 enzymes (e.g., P450 2E1) in hepatocytes to form reactive radical metabolites or oxidants that cause lipid peroxidation and cytotoxicity.

  15. Neovestitol, an isoflavonoid isolated from Brazilian red propolis, reduces acute and chronic inflammation: involvement of nitric oxide and IL-6

    Science.gov (United States)

    Franchin, Marcelo; Colón, David F.; da Cunha, Marcos G.; Castanheira, Fernanda V. S.; Saraiva, André L. L.; Bueno-Silva, Bruno; Alencar, Severino M.; Cunha, Thiago M.; Rosalen, Pedro L.

    2016-01-01

    Isoflavonoids have been largely studied due to their distinct biological activities identified thus far. Herein, we evaluated the activity of neovestitol, an isoflavonoid isolated from Brazilian red propolis, in acute and chronic inflammation. As for acute inflammation, we found that neovestitol reduced neutrophil migration, leukocyte rolling and adhesion, as well as expression of ICAM-1 in the mesenteric microcirculation during lipopolysaccharide-induced acute peritonitis. No changes were observed in the levels of TNF-α, CXCL1/KC and CXCL2/MIP-2 upon pretreatment with neovestitol. The administration of an inducible nitric oxide synthase (iNOS) inhibitor abolished the inhibitory effects of neovestitol in neutrophil migration and ICAM-1 expression. Nitrite levels increased upon treatment with neovestitol. No effects of neovestitol were observed on the chemotaxis of neutrophils in vitro. As for chronic inflammation, neovestitol also reduced the clinical score and joint damage in a collagen-induced arthritis model. There was no change in the frequency of IL-17-producing TCD4+ cells. In addition, pretreatment with neovestitol reduced the levels of IL-6. These results demonstrate a potential anti-inflammatory activity of neovestitol, which may be useful for therapeutic purposes and/or as a nutraceutical. PMID:27819273

  16. An unusual case of isolated, serial metastases of gallbladder carcinoma involving the chest wall, axilla, breast and lung parenchyma

    Directory of Open Access Journals (Sweden)

    Matthew J. Iott

    2013-02-01

    Full Text Available In the English literature, only 9 cases of adenocarcinoma of the gallbladder with cutaneous metastasis have been reported so far. One case of multiple cutaneous metastases along with deposits in the breast tissue has been reported. We present a case of incidental metastatic gallbladder carcinoma with no intra-abdominal disease presenting as a series of four isolated cutaneous right chest wall, axillary nodal, breast and pulmonary metastases following resection and adjuvant chemoradiation for her primary tumor. In spite of the metastatic disease coupled with the aggressive nature of the cancer, this patient reported that her energy level had returned to baseline with a good appetite and a stable weight indicating a good performance status and now is alive at 25 months since diagnosis. Her serially-presented, oligometastatic diseases were well-controlled by concurrent chemoradiation and stereotactic radiation therapy. We report this case study because of its rarity and for the purpose of complementing current literature with an additional example of cutaneous metastasis from adenocarcinoma of the gallbladder.

  17. Neovestitol, an isoflavonoid isolated from Brazilian red propolis, reduces acute and chronic inflammation: involvement of nitric oxide and IL-6.

    Science.gov (United States)

    Franchin, Marcelo; Colón, David F; da Cunha, Marcos G; Castanheira, Fernanda V S; Saraiva, André L L; Bueno-Silva, Bruno; Alencar, Severino M; Cunha, Thiago M; Rosalen, Pedro L

    2016-11-07

    Isoflavonoids have been largely studied due to their distinct biological activities identified thus far. Herein, we evaluated the activity of neovestitol, an isoflavonoid isolated from Brazilian red propolis, in acute and chronic inflammation. As for acute inflammation, we found that neovestitol reduced neutrophil migration, leukocyte rolling and adhesion, as well as expression of ICAM-1 in the mesenteric microcirculation during lipopolysaccharide-induced acute peritonitis. No changes were observed in the levels of TNF-α, CXCL1/KC and CXCL2/MIP-2 upon pretreatment with neovestitol. The administration of an inducible nitric oxide synthase (iNOS) inhibitor abolished the inhibitory effects of neovestitol in neutrophil migration and ICAM-1 expression. Nitrite levels increased upon treatment with neovestitol. No effects of neovestitol were observed on the chemotaxis of neutrophils in vitro. As for chronic inflammation, neovestitol also reduced the clinical score and joint damage in a collagen-induced arthritis model. There was no change in the frequency of IL-17-producing TCD4+ cells. In addition, pretreatment with neovestitol reduced the levels of IL-6. These results demonstrate a potential anti-inflammatory activity of neovestitol, which may be useful for therapeutic purposes and/or as a nutraceutical.

  18. Susceptibility pattern of some clinical bacterial isolates to selected antibiotics and disinfectants.

    Science.gov (United States)

    Ogbulie, Jude N; Adieze, Ifechukwu E; Nwankwo, Ngozi C

    2008-01-01

    The antibacterial activities of five antibiotics, three brands of Ofloxacins (Obenasin, Floxavid and Drovid) and two brands of Ciprofloxacins (Uroxin and Siprosan), and five commonly used disinfectants (Lysol, Dettol, Purit, Roberts and Wex-cide) against Staphylococcus aureus, Escherichia coli, Proteus spp., Pseudomonas aeruginosa, Streptococcus spp. and Bacillus spp. were investigated. The growth inhibitory effect of both the antibiotics and disinfectants were determined using paper disk diffusion method and well-in-agar technique respectively. The highest mean zone of growth inhibition (19.3 mm) was given by Drovid on Streptococcus spp., while the smallest (7.0 mm) was by Floxavid on P. aeruginosa. Lysol had the highest mean zone of growth inhibition (18.0 mm) on Streptococcus spp. while P. aeruginosa and Bacillus spp. had no zone of growth inhibition with Roberts at 100-fold dilution. All the isolates were also resistant to Wex-cide. The test organisms were found to be significantly susceptible to the routinely used antimicrobials tested. However, there is the need for continuous surveillance for the detection of emerging resistance pattern.

  19. Isolation of biosurfactant-producing marine bacteria and characteristics of selected biosurfactant

    Directory of Open Access Journals (Sweden)

    Kulnaree Phetrong

    2007-05-01

    Full Text Available Biosurfactant-producing marine bacteria were isolated from oil-spilled seawater collected from harbors and docks in Songkhla Province, Thailand. Haemolytic activity, emulsification activity toward nhexadecane,emulsion of weathered crude oil, drop collapsing test as well as oil displacement test were used to determine biosurfactant producing activity of marine bacteria. Among two-hundred different strains, 40strains exhibited clear zone on blood agar plates. Only eight strains had haemolytic activity and were able to emulsify weathered crude oil in marine broth during cultivation. Eight strains named SM1-SM8 wereidentified by 16S rRNA as Myroides sp. (SM1; Vibrio paraheamolyticus (SM2; Bacillus subtilis (SM3; Micrococcus luteus (SM4; Acinetobacter anitratus (SM6; Vibrio paraheamolyticus (SM7 and Bacilluspumilus (SM8. However, SM5 could not be identified. Strain SM1 showed the highest emulsification activity against weathered crude oil, by which the oil was emulsified within 24 h of cultivation. In addition, strainSM1 exhibited the highest activity for oil displacement test and emulsification test toward n-hexadecane. The emulsification activity against n-hexadecane of crude extract of strain SM1 was stable over a broadrange of temperature (30-121oC, pH (5-12 and salt concentration (0-9% NaCl, whereas CaCl2 showed an adverse effect on emulsifying activity.

  20. Selective Inhibition of Bakuchicin Isolated from Psoralea corylifolia on CYP1A in Human Liver Microsomes

    Directory of Open Access Journals (Sweden)

    Sun Joo Kim

    2016-01-01

    Full Text Available Bakuchicin is a furanocoumarin isolated from Psoralea corylifolia and shows several biological activities. Although there have been studies on the biological effects of bakuchicin, its modulation potency of CYP activities has not been previously investigated. Here, we investigated the inhibitory effects of bakuchicin on the activities of CYP isoforms by using a cocktail of probe substrates in pooled human liver microsomes (HLMs and human recombinant cDNA-expressed CYP. Bakuchicin strongly inhibited CYP1A-mediated phenacetin O-deethylation with an IC50 value of 0.43 μM in HLMs. It was confirmed by human recombinant cDNA-expressed CYP1A1 and CYP1A2 with a Ki value of 0.11 μM and 0.32 μM, respectively. A Lineweaver-Burk plot indicated that the inhibition mechanism of bakuchicin was competitive inhibition. Overall, this is the first study to investigate the potential CYP1A1 and CYP1A2 inhibition associated with bakuchicin and to report its competitive inhibitory effects on HLMs.

  1. Selection and evaluation of thermal criteria for a geologic waste isolation facility in salt

    Energy Technology Data Exchange (ETDEWEB)

    1976-09-01

    Previous design efforts for geologic waste isolation facilities in bedded salt developed several limiting temperature conditions, or thermal criteria, for use in parametric studies. These criteria, along with other design parameters, must assure that the temperature variations that occur do not adversely affect operating personnel and equipment during normal operations as well as assure containment and environmental integrity. The goals of the present study are to review past analyses of thermal criteria, determine the factors that should be considered in defining thermal criteria, suggest appropriate procedures for determining thermal criteria and suggest additional experimental and computational efforts required to adequately determine thermal criteria. The approach taken was to divide consideration of the system into four categories of thermal effects and four phases of facility operation. The categories were effects on the canister and its immediate environs, effects on the operating environment experienced by personnel and equipment, effects on storage room integrity, and effects on the biosphere. The phases of facility operation were loading, storage, retrieval, and disposal. Each of the categories was further subdivided into several aspects for detailed consideration of thermal effects through the four operating phases. This comprehensive approach was taken to ensure that all conceivable thermal effects were included, and to demonstrate a systematic approach for use in developing thermal criteria for other geologic media.

  2. Identification and Isolation of Brucella suis Virulence Genes Involved in Resistance to the Human Innate Immune System▿

    Science.gov (United States)

    Liautard, Janny; Ouahrani-Bettache, Safia; Jubier-Maurin, Véronique; Lafont, Virginie; Köhler, Stephan; Liautard, Jean-Pierre

    2007-01-01

    Brucella strains are facultative intracellular pathogens that induce chronic diseases in humans and animals. This observation implies that Brucella subverts innate and specific immune responses of the host to develop its full virulence. Deciphering the genes involved in the subversion of the immune system is of primary importance for understanding the virulence of the bacteria, for understanding the pathogenic consequences of infection, and for designing an efficient vaccine. We have developed an in vitro system involving human macrophages infected by Brucella suis and activated syngeneic γ9δ2 T lymphocytes. Under these conditions, multiplication of B. suis inside macrophages is only slightly reduced. To identify the genes responsible for this reduced sensitivity, we screened a library of 2,000 clones of transposon-mutated B. suis. For rapid and quantitative analysis of the multiplication of the bacteria, we describe a simple method based on Alamar blue reduction, which is compatible with screening a large library. By comparing multiplication inside macrophages alone and multiplication inside macrophages with activated γ9δ2 T cells, we identified four genes of B. suis that were necessary to resist to the action of the γ9δ2 T cells. The putative functions of these genes are discussed in order to propose possible explanations for understanding their exact role in the subversion of innate immunity. PMID:17709411

  3. Antimicrobial activity, toxicity and selectivity index of two biflavonoids and a flavone isolated from Podocarpus henkelii (Podocarpaceae) leaves.

    Science.gov (United States)

    Bagla, Victor P; McGaw, Lyndy J; Elgorashi, Esam E; Eloff, Jacobus N

    2014-10-08

    Different parts of Podocarpus henkelii have been used in many cultures around the world to treat ailments such as cholera, stomach diseases, rheumatism, cancer, canine distemper in dogs and gall sickness in cattle. The aim of this study was to evaluate the biological activity and toxicity of isolated compounds from Podocarpus henkelii after an earlier study indicated a promising activity in crude extracts against viral pathogens of veterinary importance. The antibacterial and antifungal activity of two biflavonoids 7, 4', 7", 4"'-tetramethoxy amentoflavone (TMA), isoginkgetin (IGG) and podocarpus flavone-A (PFA) isolated from the leaves of Podocarpus henkelii were determined using a serial microplate dilution method with tetrazolium violet as growth indicator. The cytotoxicity of compounds TMA and IGG were determined on different cell types using a tetrazolium-based colorimetric cellular assay (MTT). The Ames test was used to determine their mutagenic activities. TMA had reasonable antifungal activity against Aspergillus fumigatus (MIC = 30 μg/ml). IGG had a wide spectrum of activity against four bacterial and two fungal pathogens with much higher selectivity index values obtained for A. fumigatus and Cryptococcus neoformans (SI > 30). PFA had a broad spectrum of activity against Enterococcus faecalis and Pseudomonas aeruginosa (SI > 15) and less activity against the two fungal pathogens. In both the cytotoxicity assays and Ames mutagenicity test using Salmonella typhimurium strains TA98 and TA100, TMA and IGG had no deleterious effect on the different cell types and did not induce mutations in the Ames test. Although the antimicrobial activities of the isolated compounds were not that exciting, the compounds had no cytotoxic activity at the highest concentration (1000 μg/ml) tested against all three cell lines. IGG was the most active against E. coli, S. aureus, A. fumigatus and C. neoformans, exhibiting both antibacterial and antifungal activity with good

  4. Hairy Cell Leukemia Presenting with Isolated Skeletal Involvement Successfully Treated by Radiation Therapy and Cladribine: A Case Report and Review of the Literature

    Directory of Open Access Journals (Sweden)

    Ipek Yonal-Hindilerden

    2015-01-01

    Full Text Available We describe an unusual case of hairy cell leukemia (HCL in a 55-year-old male presenting with isolated skeletal disease as the initial manifestation without abnormal peripheral blood counts, bone marrow involvement, or splenomegaly. To the best of our knowledge, there have been only two previous reports of a similar case. The patient presented with pain in the right femur. Anteroposterior radiographs of both femurs revealed mixed lytic-sclerotic lesions. PET scan showed multiple metastatic lesions on axial skeleton, pelvis, and both femurs. Histopathological examination of the bone biopsy revealed an infiltrate of HCL. Localized radiation therapy to both proximal femurs and subsequently 4 weeks later, a 7-day course of 0.1 mg/kg/day cladribine provided complete remission with relief of symptoms and resolution of bone lesions. We addressed the manifestations and management of HCL patients with skeletal involvement.

  5. Selected lactic acid-producing bacterial isolates with the capacity to reduce Salmonella translocation and virulence gene expression in chickens.

    Directory of Open Access Journals (Sweden)

    Xiaojian Yang

    Full Text Available BACKGROUND: Probiotics have been used to control Salmonella colonization/infection in chickens. Yet the mechanisms of probiotic effects are not fully understood. This study has characterized our previously-selected lactic acid-producing bacterial (LAB isolates for controlling Salmonella infection in chickens, particularly the mechanism underlying the control. METHODOLOGY/PRINCIPAL FINDINGS: In vitro studies were conducted to characterize 14 LAB isolates for their tolerance to low pH (2.0 and high bile salt (0.3-1.5% and susceptibility to antibiotics. Three chicken infection trials were subsequently carried out to evaluate four of the isolates for reducing the burden of Salmonella enterica serovar Typhimurium in the broiler cecum. Chicks were gavaged with LAB cultures (10(6-7 CFU/chick or phosphate-buffered saline (PBS at 1 day of age followed by Salmonella challenge (10(4 CFU/chick next day. Samples of cecal digesta, spleen, and liver were examined for Salmonella counts on days 1, 3, or 4 post-challenge. Salmonella in the cecum from Trial 3 was also assessed for the expression of ten virulence genes located in its pathogenicity island-1 (SPI-1. These genes play a role in Salmonella intestinal invasion. Tested LAB isolates (individuals or mixed cultures were unable to lower Salmonella burden in the chicken cecum, but able to attenuate Salmonella infection in the spleen and liver. The LAB treatments also reduced almost all SPI-1 virulence gene expression (9 out of 10 in the chicken cecum, particularly at the low dose. In vitro treatment with the extracellular culture fluid from a LAB culture also down-regulated most SPI-1 virulence gene expression. CONCLUSIONS/SIGNIFICANCE: The possible correlation between attenuation of Salmonella infection in the chicken spleen and liver and reduction of Salmonella SPI-1 virulence gene expression in the chicken cecum by LAB isolates is a new observation. Suppression of Salmonella virulence gene expression in

  6. Selection and identification of fungi isolated from sugarcane bagasse and their application for phenanthrene removal from soil.

    Science.gov (United States)

    Cortés-Espinosa, D V; Fernández-Perrino, F J; Arana-Cuenca, A; Esparza-García, F; Loera, O; Rodríguez-Vázquez, R

    2006-01-01

    This work investigated the identification and selection of fungi isolated from sugarcane bagasse and their application for phenanthrene (Phe) removal from soil. Fungi were identified by PCR amplification of ITS regions as Aspergillus terrus, Aspergillus fumigatus and Aspergillus niger, Penicillium glabrum and Cladosporium cladosporioides. A primary selection of fungi was accomplished in plate, considering Phe tolerance of every strain in two different media: potato dextrose agar (PDA) and mineral medium (MM). The radial extension rate (r(r)) in PDA exhibited significant differences (p<0.05) at 200 and 400 ppm of Phe. A secondary selection of A. niger, C. cladosporoides, and P. glabrum sp. was achieved based on their tolerance to 200, 400, 600 and 800 ppm of Phe, in solid culture at a sugarcane bagasse/contaminated soil ratio of 95:5, in Toyamas, Czapeck and Wunder media. Under these conditions, a maximum (70%) Phe removal by A. niger was obtained. In addition C. cladosporioides and A. niger were able to remove high (800 ppm) Phe concentrations.

  7. Analysis of polymorphisms and selective pressures on ama1 gene in Plasmodium knowlesi isolates from Sabah, Malaysia

    Indian Academy of Sciences (India)

    CHUEN YANG CHUA; PINGCHIN LEE; TIEK YING LAU

    2017-09-01

    The apical membrane antigen-1 (AMA-1) of Plasmodium spp. is a merozoite surface antigen that is essential for the recognition and invasion of erythrocytes. Polymorphisms occurring in this surface antigen will cause major obstacles in developing effective malaria vaccines based on AMA-1. The objective of this study was to characterize ama1 gene in Plasmodium knowlesi isolates from Sabah. DNA was extracted from blood samples collected from Keningau, Kota Kinabalu and Kudat. The Pkama1 gene was amplified using nested PCR and subjected to bidirectional sequencing. Analysis of DNA sequence revealed that most of the nucleotide polymorphisms were synonymous and concentrated in domain I of PkAMA-1. Forteen haplotypes were identified based on amino acid variations and haplotype K5 was the most common haplotype. dN/dS ratios implied that purifying selection was prevalent in Pkama1 gene. Fu and Li’s D and F values further provided evidence of negative selection acting on domain II ofPkama1. Lownucleotide diversitywas also detected for the Pkama1 sequences,which is similar to reports on Pkama1 from Peninsular Malaysia and Sarawak. The presence of purifying selection and low nucleotide diversity indicated that domain II of Pkama1 can be used as a target for vaccine development.

  8. Analysing one isolated single walled carbon nanotube in the near-field domain with selective nanovolume Raman spectroscopy.

    Science.gov (United States)

    Atalay, Han; Lefrant, Serge

    2004-09-01

    In this paper, we describe a new method to the selective nanovolume analysing of one isolated single walled carbon nanotube (SWNT). This concept is based on actually available imaging micro-spectrometry systems for working in near-field domain combined with a stigmatic solid immersion lens. This combination of different analytical methods, and modified and configured equipment entitles us to expand the functionality toward a three-dimensional (3D) nanovolume Raman mapping and photoluminescence intensity with a possible discrimination in polarization, as well as photoluminescence decaytime constant mapping with their unique combination. Subsequently, selective spectra can be acquired from the same location on the samples. By spectrally selecting a SWNT, we registered the spatial distribution of the emitted photons in x, y, z vectors to determine the position of a SWNT in the near-field domain. For the SWNTs that are localized with an accuracy better than 18 nm in the x, y and unity throughput. This near-field capability is applied to resolve local variations unambiguously in the Raman spectrum along one single SWNT. Finally, in this paper, we report what we believe to be the first evidence of Raman mapping and 3D real optical imaging of carbon nanotubes with near-field resolution.

  9. Brain activation in teenagers with isolated spelling disorder during tasks involving spelling assessment and comparison of pseudowords. fMRI study.

    Science.gov (United States)

    Borkowska, Aneta Rita; Francuz, Piotr; Soluch, Paweł; Wolak, Tomasz

    2014-10-01

    The present study aimed at defining the specific traits of brain activation in teenagers with isolated spelling disorder in comparison with good spellers. fMRI examination was performed where the subject's task involved taking a decision 1/whether the visually presented words were spelled correctly or not (the orthographic decision task), and 2/whether the two presented letters strings (pseudowords) were identical or not (the visual decision task). Half of the displays showing meaningful words with an orthographic difficulty contained pairs with both words spelled correctly, and half of them contained one misspelled word. Half of the pseudowords were identical, half of them were not. The participants of the study included 15 individuals with isolated spelling disorder and 14 good spellers, aged 13-15. The results demonstrated that the essential differences in brain activation between teenagers with isolated spelling disorder and good spellers were found in the left inferior frontal gyrus, left medial frontal gyrus and right cerebellum posterior lobe, i.e. structures important for language processes, working memory and automaticity of behaviour. Spelling disorder is not only an effect of language dysfunction, it could be a symptom of difficulties in learning and automaticity of motor and visual shapes of written words, rapid information processing as well as automating use of orthographic lexicon. Copyright © 2013 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.

  10. The use of titanium dioxide micro-columns to selectively isolate phosphopeptides from proteolytic digests

    DEFF Research Database (Denmark)

    Thingholm, Tine E; Larsen, Martin R

    2009-01-01

    Titanium dioxide has very high affinity for phosphopeptides and it has become an efficient alternative to already existing methods for phosphopeptide enrichment from complex samples. Peptide loading in a highly acidic environment in the presence of 2,5-dihydroxybenzoic acid (DHB), phthalic acid......, or glycolic acid has been shown to improve selectivity significantly by reducing unspecific binding from nonphosphorylated peptides. The enriched phosphopeptides bound to the titanium dioxide are subsequently eluted from the micro-column using an alkaline buffer. Titanium dioxide chromatography is extremely...... tolerant towards most buffers used in biological experiments. It is highly robust and as such it has become one of the methods of choice in large-scale phospho-proteomics. Here we describe the protocol for phosphopeptide enrichment using titanium dioxide chromatography followed by desalting...

  11. Isolation, Characterization, and Selection of Molds Associated to Fermented Black Table Olives

    DEFF Research Database (Denmark)

    Bavaro, Simona L.; Susca, Antonia; Frisvad, Jens Christian

    2017-01-01

    Table olives are one of the most important fermented food in the Mediterranean countries. Apart from lactic acid bacteria and yeasts that mainly conduct the olive fermentation, molds can develop on the brine surface, and can have either deleterious or useful effects on this process. From the food...... with biotechnological interest. These are putative candidates for future studies as autochthonous co-starters with yeasts and lactic acid bacteria for black table olive production....... (biogenic amines and secondary metabolites, including mycotoxins) characterization. Combining both technological (presence of desired and absence of undesired enzymatic activities) and safety aspects (no or low production of biogenic amines and regulated mycotoxins), it was possible to select six strains...

  12. Selective inactivation by 21-chlorinated steroids of rabbit liver and adrenal microsomal cytochromes P-450 involved in progesterone hydroxylation.

    Science.gov (United States)

    Halpert, J; Jaw, J Y; Balfour, C; Mash, E A; Johnson, E F

    1988-08-01

    The inactivation by 21-chlorinated steroids of rabbit liver cytochromes P-450 involved in the hydroxylation of progesterone has been investigated in intact microsomes encompassing two phenotypes of 21-hydroxylase activity, two phenotypes of 16 alpha-hydroxylase activity, and three phenotypes of 6 beta-hydroxylase activity. In liver microsomes from outbred New Zealand White male rabbits exhibiting a high content of cytochrome P-450 1, 21,21-dichloropregnenolone caused a time- and NADPH-dependent loss of 21-hydroxylase activity. This loss of activity exhibited a number of characteristics of mechanism-based inactivation, including irreversibility, saturation with increasing inhibitor concentrations, and protection by substrate, and was also documented with purified P-450 1 in a reconstituted system. 21,21-Dichloropregnenolone caused no time-dependent loss of 6 beta-hydroxylase activity in microsomes from the New Zealand White rabbits or from control or rifampicin-treated rabbits of the inbred B/J strain. In contrast, in the microsomes from the B/J rabbits, some inactivation of the 16 alpha-hydroxylase was observed (k = 0.04 min-1), regardless of the rifampicin treatment. The other two compounds tested, 21-chloropregnenolone and 21,21-dichloroprogesterone, were less effective than the dichloropregnenolone as inactivators of cytochrome P-450 1. On the other hand, 21,21-dichloroprogesterone, but not 21,21-dichloropregneolone, caused a rapid time-dependent loss of 21-hydroxylase activity in rabbit adrenal microsomes. The results indicate that the introduction of a dichloromethyl group into a substrate bearing a methyl group normally hydroxylated by only one or a few forms of cytochrome P-450 may be a rational means of designing selective inhibitors of the enzyme.

  13. Joint Relationships between Civic Involvement, Higher Education, and Selected Personal Characteristics among Adults in the United States

    OpenAIRE

    Blanks, Felica B. Wooten

    2000-01-01

    American democracy fosters the common good of society by allowing citizen involvement in government. Sustaining American democracy depends on civic involvement among citizens. Civic involvement, which consists of citizens' informed involvement in government, politics, and community life, is a desired behavior among adult citizens in the United States and it is a desired outcome of higher education. However, people in the latter part of the twentieth century have questioned the extent to which...

  14. Joint Relationships between Civic Involvement, Higher Education, and Selected Personal Characteristics among Adults in the United States

    OpenAIRE

    Blanks, Felica B. Wooten

    2000-01-01

    American democracy fosters the common good of society by allowing citizen involvement in government. Sustaining American democracy depends on civic involvement among citizens. Civic involvement, which consists of citizens' informed involvement in government, politics, and community life, is a desired behavior among adult citizens in the United States and it is a desired outcome of higher education. However, people in the latter part of the twentieth century have questioned the extent to which...

  15. Mepivacaine-induced contraction involves phosphorylation of extracellular signal-regulated kinase through activation of the lipoxygenase pathway in isolated rat aortic smooth muscle.

    Science.gov (United States)

    Lee, Hyo Min; Ok, Seong-Ho; Sung, Hui-Jin; Eun, So Young; Kim, Hye Jung; Lee, Soo Hee; Kang, Sebin; Shin, Il-Woo; Lee, Heon Keun; Chung, Young-Kyun; Choi, Mun-Jeoung; Bae, Sung Il; Sohn, Ju-Tae

    2013-04-01

    Mepivacaine is an aminoamide local anesthetic with an intermediate duration that intrinsically produces vasoconstriction both in vivo and in vitro. This study investigated the arachidonic acid metabolic pathways involved in mepivacaine-induced contraction, and elucidated the associated cellular mechanism with a particular focus on extracellular signal-regulated kinase (ERK) in endothelium-denuded rat aorta. Isolated rat thoracic aortic rings were suspended for isometric tension recording. Cumulative mepivacaine concentration-response curves were generated in the presence or absence of the following inhibitors: quinacrine dihydrochloride, nordihydroguaiaretic acid, phenidone, AA-861, indomethacin, NS-398, SC-560, fluconazole, PD 98059, and verapamil. Mepivacaine-induced ERK phosphorylation, 5-lipoxygenase (5-LOX) expression, and cyclooxygenase (COX)-2 expression in rat aortic smooth muscle cells were detected by Western blot analysis in the presence or absence of inhibitors. Mepivacaine produced tonic contraction in isolated endothelium-denuded rat aorta. Quinacrine dihydrochloride, nordihydroguaiaretic acid, phenidone, AA-861, NS-398, PD 98059, and verapamil attenuated mepivacaine-induced contraction in a concentration-dependent manner. However, fluconazole had no effect on mepivacaine-induced contraction. PD 98059, quinacrine dihydrochloride, nordihydroguaiaretic acid, AA-861, phenidone, and indomethacin attenuated mepivacaine-induced ERK phosphorylation. Mepivacaine upregulated 5-LOX and COX-2 expression. These results suggest that mepivacaine-induced contraction involves ERK activation, which is primarily mediated by the 5-LOX pathway and in part by the COX-2 pathway.

  16. A select bibliography with abstracts of reports related to Waste Isolation Pilot Plant geotechnical studies (1972--1990)

    Energy Technology Data Exchange (ETDEWEB)

    Powers, D.W. [Powers (Dennis W.), Anthony, TX (United States); Martin, M.L. [International Technology, Inc., Las Vegas, NV (United States)

    1993-08-01

    This select bibliography contains 941 entries. Each bibliographic entry contains the citation of a report, conference paper, or journal article containing geotechnical information about the Waste Isolation Pilot Plant (WIPP). The entries cover the period from 1972, when investigation began for a WIPP Site in southeastern New Mexico, through December 1990. Each entry is followed by an abstract. If an abstract or suitable summary existed, it has been included; 316 abstracts were written for other documents. For some entries, an annotation has been provided to clarify the abstract, comment on the setting and significance of the document, or guide the reader to related reports. An index of key words/phrases is included for all entries.

  17. Identification of Highly Potent and Selective α-Glucosidase Inhibitors with Antiglycation Potential, Isolated from Rhododendron arboreum

    Directory of Open Access Journals (Sweden)

    Rabia Raza

    2015-01-01

    Full Text Available This study explored antidiabetic potential of eight known pure compounds, isolated from the bark of Rhododendron arboreum. Invitro studies of these compounds against α and β-glucosidases revealed them as very potent and selective inhibitors of α-glucosidase. Compound 7 (3-O-acetylursolic acid was found to be the most potent inhibitor of α-glucosidase with 3.3±0.1µM IC 50 value which was many folds higher than standard inhibitor acarbose. Antiglycation studies of compounds showed that all compounds were also very active antiglycation agents. The studied biological properties of these compounds suggest that they are therapeutically interesting and important tools for treatment of diabetes.

  18. [Differences between species involved and fluoroquinolone resistance patterns of strains isolated from bacteriuria according to nosocomial, health-related or community-acquired onset].

    Science.gov (United States)

    Tassain, Jérome; N'Guyen, Yohan; Batalla, Anne-Sophie; Duval, Véronique; Guillard, Thomas; De Champs, Christophe; Strady, Christophe

    2012-12-01

    To describe and to compare species and antibiotics resistance patterns of bacteria involved among bacteriuria from hospital and city laboratories and among health-related and community-acquired bacteriuria. Epidemiologic transversal study conducted among Bacteriology laboratories of University Hospital (UH) and the whole city of Reims, during the week 21 to 26 June 2010. A standardized investigation form was completed after telephonical interview with the prescriber. One hundred and eighty-nine strains have been isolated among 179 urocultures. One hundred and seven strains were isolated in city laboratories and 82 in UH laboratory. Strains were community-acquired, health-related and nosocomial in 136, 22 and 31 cases, respectively. More Gram positive bacteria and ofloxacin resistant strains were isolated among UH strains (P=0.001 and P=0.015, respectively) and among health-related strains (P=0.01 and P=0.003, respectively). When analysis was restricted only to Enterobacteriaceae or to Escherichia coli, the ofloxacin resistance rate was no more elevated among health-related or UH strains. Ofloxacin resistant Enterobacteriaceae were more frequently resistant to all other classes of antibiotics except nitrofurans. Strains isolated in health-related bacteriuria are more frequently ofloxacin resistant principally because of the greater proportion of Gram positive bacteria and because of a non-significant higher ofloxacin resistance rate among Enterobacteriaceae. Numerous studies only focus on Enterobacteriaceae, and the data from our study need to be confirmed on larger samples, in order to validate the predictive value of health-related bacteriuria for ofloxacin resistance. Copyright © 2012. Published by Elsevier Masson SAS.

  19. Escherichia coli and selected veterinary and zoonotic pathogens isolated from environmental sites in companion animal veterinary hospitals in southern Ontario

    Science.gov (United States)

    Murphy, Colleen P.; Reid-Smith, Richard J.; Boerlin, Patrick; Weese, J. Scott; Prescott, John F.; Janecko, Nicol; Hassard, Lori; McEwen, Scott A.

    2010-01-01

    Hospital-based infection control in veterinary medicine is emerging and the role of the environment in hospital-acquired infections (HAI) in veterinary hospitals is largely unknown. This study was initiated to determine the recovery of Escherichia coli and selected veterinary and zoonotic pathogens from the environments of 101 community veterinary hospitals. The proportion of hospitals with positive environmental swabs were: E. coli — 92%, Clostridium difficile — 58%, methicillin-resistant Staphylococcus aureus (MRSA) — 9%, CMY-2 producing E. coli — 9%, methicillin-resistant Staphylococcus pseudintermedius — 7%, and Salmonella — 2%. Vancomycin-resistant Enterococcus spp., canine parvovirus, and feline calicivirus were not isolated. Prevalence of antimicrobial resistance in E. coli isolates was low. Important potential veterinary and human pathogens were recovered including Canadian epidemic strains MRSA-2 and MRSA-5, and C. difficile ribotype 027. There is an environmental reservoir of pathogens in veterinary hospitals; therefore, additional studies are required to characterize risk factors associated with HAI in companion animals, including the role of the environment. PMID:21119862

  20. Prevalence and Bacterial Isolates of Mastitis in Dairy Farms in Selected Districts of Eastern Harrarghe Zone, Eastern Ethiopia.

    Science.gov (United States)

    Zeryehun, Tesfaheywet; Abera, Gerema

    2017-01-01

    The study was conducted from November 2015 to April 2016 to estimate the prevalence of clinical and subclinical mastitis in lactating cows, to assess the associated risk factors, and to isolate the major bacterial pathogens in dairy farms in selected district of Eastern Harrarghe Zone, Eastern Ethiopia. The study was carried out in 384 dairy cows based on data collection, farm visit, animal examination, California mastitis test (CMT), and isolation bacterial pathogens using standard techniques. In the present study the overall mastitis at cow level was 247 (64.3%). The prevalence of clinical and subclinical mastitis and quarter level prevalence for clinical and subclinical mastitis were 12.5% and 51.8% at cow level and 10.7% and 46.4% at quarter level, respectively. Clinically, 101 (6.6%) quarters which belong to 75 (19.5%) animals were found to be with blind teat. In the present study prevalence of mastitis was significantly associated with parity and age (p mastitis particularly the subclinical mastitis was major problem of dairy cows in the area and hence warrants serious attention.

  1. Poly-γ-Glutamic Acids Contribute to Biofilm Formation and Plant Root Colonization in Selected Environmental Isolates of Bacillus subtilis.

    Science.gov (United States)

    Yu, Yiyang; Yan, Fang; Chen, Yun; Jin, Christopher; Guo, Jian-Hua; Chai, Yunrong

    2016-01-01

    Bacillus subtilis is long known to produce poly-γ-glutamic acids (γ-PGA) as one of the major secreted polymeric substances. In B. subtilis, the regulation of γ-PGA production and its physiological role are still unclear. B. subtilis is also capable of forming structurally complex multicellular communities, or biofilms, in which an extracellular matrix consisting of secreted proteins and polysaccharides holds individual cells together. Biofilms were shown to facilitate B. subtilis-plant interactions. In this study, we show that different environmental isolates of B. subtilis, all capable of forming biofilms, vary significantly in γ-PGA production. This is possibly due to differential regulation of γ-PGA biosynthesis genes. In many of those environmental isolates, γ-PGA seems to contribute to robustness and complex morphology of the colony biofilms, suggesting a role of γ-PGA in biofilm formation. Our evidence further shows that in selected B. subtilis strains, γ-PGA also plays a role in root colonization by the bacteria, pinpointing a possible function of γ-PGA in B. subtilis-plant interactions. Finally, we found that several pathways co-regulate both γ-PGA biosynthesis genes and genes for the biofilm matrix in B. subtilis, but in an opposing fashion. We discussed potential biological significance of that.

  2. High-removal selectivity through interaction between polyacrylamide and SiO2 film in poly isolation chemical mechanical planarization.

    Science.gov (United States)

    Kim, Ye-Hwan; Lee, Kee-June; Park, Jea-Gun; Paik, Ungyu

    2009-06-01

    The interaction between polyacrylamide (PAM) and SiO2 film was investigated in order to elucidate the removal polycrystalline silicon (poly Si) to SiO2 selectivity in poly isolation chemical mechanical planarization (CMP). The hydrophilic characteristics of poly Si and SiO2 were analyzed by the X-ray photoelectron spectroscopy (XPS) and contact angle measurement. The surface of SiO2 is more hydrophilic than that of poly Si due to the siloxane (triple bond Si-O-Si triple bond) bonding. The adsorption behavior of PAM on poly Si and SiO2 film was determined by adsorption isotherms and force measurements using atomic force microscopy (AFM). Interaction between siloxane bonding of SiO2 film and the amine group along the backbone of PAM results in the adsorption of PAM on SiO2 film. Consequently, the passivation layer of PAM on the SiO2 film prevented abrasives from approaching the surface of SiO2 film, which led to suppression of the removal rate of SiO2 film from 82 to 12 A/min in poly isolation CMP process.

  3. Public Involvement in Repository Site Selection for Nuclear Waste: Towards a more Dynamic View in Decision-Making Processes

    Energy Technology Data Exchange (ETDEWEB)

    Kruetli, Pius; Stauffacher, Michael; Flueeler, Thomas; Scholz, Roland W. [ETH Zuerich (Switzerland). lnst. for Human-Environment Systems (HES)

    2006-09-15

    This paper discusses possibilities of public involvement in radioactive waste management. A general overview of the radioactive waste issue is presented referring to a proposed model of the respective decision-making process. Based on the well known participation ladder by Arnstein, we differentiate various intensities of public involvement. A matrix with public involvement and the decision-making process is introduced and three prototypical patterns are discussed. We conclude that time frame, the level of public involvement and the mission have to be considered as well as techniques and the overarching context - all in all, a systematic and dynamic approach for public involvement is needed.

  4. Isolating GaSb membranes grown metamorphically on GaAs substrates using highly selective substrate removal etch processes

    Energy Technology Data Exchange (ETDEWEB)

    Lavrova, Olga [Univ. of New Mexico, Albuquerque, NM (United States). Dept. of Electrical and Computer Engineering. Center for High Technology Materials; Balakrishnan, Ganesh [Univ. of New Mexico, Albuquerque, NM (United States). Dept. of Electrical and Computer Engineering. Center for High Technology Materials

    2017-02-24

    The etch rates of NH4OH:H2O2 and C6H8O7:H2O2 for GaAs and GaSb have been investigated to develop a selective etch for GaAs substrates and to isolate GaSb epilayers grown on GaAs. The NH4OH:H2O2 solution has a greater etch rate differential for the GaSb/GaAs material system than C6H8O7:H2O2 solution. The selectivity of NH4OH:H2O2 for GaAs/GaSb under optimized etch conditions has been observed to be as high as 11471 ± 1691 whereas that of C6H8O7:H2O2 has been measured up to 143 ± 2. The etch contrast has been verified by isolating 2 μm thick GaSb epi-layers that were grown on GaAs substrates. GaSb membranes were tested and characterized with high-resolution X-Ray diffraction (HR-XRD) and atomic force microscopy (AFM).

  5. Tigecycline-Amikacin Combination Effectively Suppresses the Selection of Resistance in Clinical Isolates of KPC-Producing Klebsiella pneumoniae.

    Science.gov (United States)

    Ni, Wentao; Wei, Chuanqi; Zhou, Chufei; Zhao, Jin; Liang, Beibei; Cui, Junchang; Wang, Rui; Liu, Youning

    2016-01-01

    By far, only tigecycline, colistin, and some aminoglycosides still show favorable in vitro activities against carbapenem-resistant Enterobacteriaceae. However, rapid emergence of resistance often occurs during long-term treatment in clinic, challenging these last resort antimicrobials. In this study, we measured mutant prevention concentration (MPC) and mutant selection window (MSW) of tigecycline, colistin and amikacin alone and in combination for clinical isolates of KPC-producing K. pneumoniae, and characterized the resistant mutants recovered. The MPC90 of 30 tested isolates for tigecycline, colistin, and amikacin were 16, >128, and 128 mg/L, respectively. The average MSW of tigecycline-amikacin, tigecycline-colistin, and amikacin-colistin combinations for four representative strains were 11.99, 200.13, and 372.38, respectively. A strong correlation was found between the MSW combination and the product of MSW of each single drug. Combinations of 1 minimal inhibitory concentration (MIC) multiple tigecycline and 1 MIC multiple amikacin could result in 1000- to 10000-fold reduction in mutational frequency relative to their individual mutational frequencies, and combinations of 1 MIC multiple amikacin and 1.5-2 MIC multiple tigecycline could successfully restrict the recovery of resistant mutants on agar plates. However, 2 MIC multiple colistin in combination with 2 MIC multiple tigecycline or amikacin merely resulted in approximately 10-fold decrease in the mutational frequency. In conclusion, this study showed tigecycline-amikacin combination could effectively suppress the selection of resistance at low concentrations compared with the colistin-tigecycline and colistin-amikacin combinations, suggesting that this combination may be useful in clinical therapy.

  6. Tigecycline-amikacin combination effectively suppresses the selection of resistance in clinical isolates of KPC-producing Klebsiella pneumoniae

    Directory of Open Access Journals (Sweden)

    Wentao Ni

    2016-08-01

    Full Text Available By far, only tigecycline, colistin and some aminoglycosides still show favorable in vitro activities against carbapenem-resistant Enterobacteriaceae. However, rapid emergence of resistance often occurs during long-term treatment in clinic, challenging these last resort antimicrobials. In this study, we measured mutant prevention concentration (MPC and mutant selection window (MSW of tigecycline, colistin and amikacin alone and in combination for clinical isolates of KPC-producing K. pneumoniae, and characterized the resistant mutants recovered. The MPC90 of 30 tested isolates for tigecycline, colistin and amikacin were 16, >128 and 128 mg/L, respectively. The average MSW of tigecycine-amikacin, tigecycline-colisitn, and amikacin-colistin combinations for 4 representative strains were 11.99, 200.13 and 372.38, respectively. A strong correlation was found between the MSWcombination and the product of MSW of each single drug. Combinations of 1MIC multiple tigecycline and 1MIC multiple amikacin could result in 1000-10000 fold reduction in mutational frequency relative to their individual mutational frequencies, and combinations of 1 MIC multiple amikacin and 1.5-2 MIC multiple tigecycline could successfully restrict the recovery of resistant mutants on agar plates. However, 2 MIC multiple colistin in combination with 2 MIC multiple tigecycline or amikacin merely resulted in approximately 10 fold decrease in the mutational frequency. In conclusion, this study showed tigecycline-amikacin combination could effectively suppress the selection of resistance at low concentrations compared with the colistin-tigecycline and colistin-amikacin combinations, suggesting that this combination may be useful in clinical therapy.

  7. Selective effect of 2',6'-dihydroxy-4'-methoxychalcone isolated from Piper aduncum on Leishmania amazonensis.

    Science.gov (United States)

    Torres-Santos, E C; Moreira, D L; Kaplan, M A; Meirelles, M N; Rossi-Bergmann, B

    1999-05-01

    2',6'-Dihydroxy-4'-methoxychalcone (DMC) was purified from the dichloromethane extract of Piper aduncum inflorescences. DMC showed significant activity in vitro against promastigotes and intracellular amastigotes of Leishmania amazonensis, with 50% effective doses of 0.5 and 24 micrograms/ml, respectively. Its inhibitory effect on amastigotes is apparently a direct effect on the parasites and is not due to activation of the nitrogen oxidative metabolism of macrophages, since the production of nitric oxide by both unstimulated and recombinant gamma interferon-stimulated macrophages was decreased rather than increased with DMC. The phagocytic activity of macrophages was functioning normally even with DMC concentrations as high as 80 micrograms/ml, as seen by electron microscopy and by the uptake of fluorescein isothiocyanate-labeled beads. Ultrastructural studies also showed that in the presence of DMC the mitochondria of promastigotes were enlarged and disorganized. Despite destruction of intracellular amastigotes, no disarrangement of macrophage organelles were observed, even at 80 micrograms of DMC/ml. These observations suggest that DMC is selectively toxic to the parasites. Its simple structure may well enable it to serve as a new lead compound for the synthesis of novel antileishmanial drugs.

  8. Detection and isolation of selected genes of interest from metagenomic libraries by a DNA microarray approach.

    Science.gov (United States)

    Pathak, Gopal P; Gärtner, Wolfgang

    2010-01-01

    A DNA microarray-based approach is described for screening metagenomic libraries for the presence of selected genes. The protocol is exemplified for the identification of flavin-binding, blue-light-sensitive biological photoreceptors (BL), based on a homology search in already sequenced, annotated genomes. The microarray carried 149 different 54-mer oligonucleotides, derived from consensus sequences of BL photoreceptors. The array could readily identify targets carrying 4% sequence mismatch, and allowed unambiguous identification of a positive cosmid clone of as little as 10 ng against a background of 25 μg of cosmid DNA. The protocol allows screening up to 1,200 library clones in concentrations as low as ca. 20 ng, each with a ca. 40 kb insert size readily in a single batch. Calibration and control conditions are outlined. This protocol, when applied to the thermophilic fraction of a soil sample, yielded the identification and functional characterization of a novel, BL-encoding gene that showed a 58% similarity to a known, BL-encoding gene from Kineococcus radiotolerans SRS30216 (similarity values refer to the respective LOV domains).

  9. Selection and Characterization of a Lysine Yielding Mutant of Corynebacterium glutamicum - a Soil Isolate from Pakistan

    Directory of Open Access Journals (Sweden)

    Habib-ur-Rehman§٭, Abdul Hameed and Safia Ahmed

    2012-01-01

    Full Text Available L-lysine is the second limiting amino acid for poultry and supplemented in broiler feed for optimal performance. Lysine can be produced by inducing mutation in glutamate producing bacteria. The study was conducted to enhance lysine production from a local strain of Corynebacterium glutamicum. The bacterium was mutated by exposure to UV. Mutants resistant to s-2-aminoethyle L-cystein (AEC and showing auxotrophy for L-homoserine were screened for lysine production qualitatively and quantitatively. A mutant showing highest production of lysine (8.2 mg/mL was selected for optimization of physical and nutritional parameters for maximum production of lysine in shake flask. An initial pH 7.6, 30˚C temperature, 300 rpm and 60 h incubation time were the optimized values of physical requirements. Cane molasses and corn starch hydrolysate were required at 15% (w/v in the fermentation media which provided around 9% total sugars to produce maximum lysine (17 to 18 mg/mL. When amonium sulphate was used at 3.5% (w/v level in molasses or corn starch hydrolysate based fermentation media, production of lysine slightly increased above 18 mg/mL. It is concluded that industrial by products like cane molasses, corn steep liquor, and corn starch hydrolysate can be used as carbon and organic nitrogen sources in fermentation medium for scale up process of lysine production and this lysine enriched broth may be used in broiler feed later. However, more potent lysine producing mutant and additional in vivo trials would be required to commercialize this product.

  10. Soft X-ray properties of a spectroscopically selected sample of interacting and isolated Seyfert galaxies

    Science.gov (United States)

    Pfefferkorn, F.; Boller, Th.; Rafanelli, P.

    2001-03-01

    We present a catalogue of ROSAT detected sources in the sample of spectroscopically selected Seyfert 1 and Seyfert 2 galaxies of Rafanelli et al. (\\cite{Rafanelli95}). The catalogue contains 102 Seyfert 1 and 36 Seyfert 2 galaxies. The identification is based on X-ray contour maps overlaid on optical images taken from the Digitized Sky Survey. We have derived the basic spectral and timing properties of the X-ray detected Seyfert galaxies. For Seyfert 1 galaxies a strong correlation between photon index and X-ray luminosity is detected. We confirm the presence of generally steeper X-ray continua in narrow-line Seyfert 1 galaxies (NLS1s) compared to broad-line Seyfert 1 galaxies. Seyfert 2 galaxies show photon indices similar to those of NLS1s. Whereas a tendency for an increasing X-ray luminosity with increasing interaction strength is found for Seyfert 1 galaxies, such a correlation is not found for Seyfert 2 galaxies. For Seyfert 1 galaxies we found also a strong correlation for increasing far-infrared luminosity with increasing interaction strength. Both NLS1s and Seyfert 2 galaxies show the highest values of far-infrared luminosity compared to Seyfert 1 galaxies, suggesting that NLS1s and Seyfert 2 galaxies host strong (circumnuclear) star formation. For variable Seyfert galaxies we present the X-ray light curves obtained from the ROSAT All-Sky Survey and from ROSAT PSPC and HRI pointed observations. Besides the expected strong short- and long-term X-ray variability in Seyfert 1 galaxies, we find indications for X-ray flux variations in Seyfert 2 galaxies. All overlays can be retrieved via CDS anonymous ftp to cdsarc.u-strasbg.fr (130.79.128.5)} or via http://cdsweb.u-strasbg.fr/cgi-bin/qcat?J/A+A/368/797

  11. Antimicrobial and selected in vitro enzyme inhibitory effects of leaf extracts, flavonols and indole alkaloids isolated from Croton menyharthii.

    Science.gov (United States)

    Aderogba, Mutalib A; Ndhlala, Ashwell R; Rengasamy, Kannan R R; Van Staden, Johannes

    2013-10-11

    Croton species are used in folk medicine in the management of infections, inflammation and oxidative stress-related diseases. In order to isolate, characterize and evaluate the bioactive constituents of Croton menyharthii Pax leaf extracts, repeated column fractionation of the ethyl acetate fraction from a 20% aqueous methanol crude extract afforded three flavonols identified by NMR (1D and 2D) spectroscopic methods as myricetrin-3-O-rhamnoside (myricetrin, 1), quercetin-3-O-rhamnoside (2) and quercetin (3) along with an indole alkaloid, (E)-N-(4-hydroxycinnamoyl)-5-hydroxytryptamine, [trans-N-(p-coumaroyl) serotonin, 4]. All the compounds are reported from the leaf extract of this plant for the first time. The crude extracts, four solvent fractions (hexane, DCM, ethyl acetate and butanol) and isolated compounds obtained from the leaves were evaluated for their inhibitory effects on selected bacteria, a fungus (Candida albicans), cyclooxygenase (COX-2), α-glucosidase and acetylcholinesterase (AChE). Amongst the compounds, quercetin (3) was the most active against Bacillus subtilis and Candida albicans while myricetrin-3-O-rhamnoside (1) and trans-N-(p-coumaroyl) serotonin (4) were the most active compounds against Escherichia coli, Klebsiella pneumonia and Staphylococcus aureus. The inhibitory activity of myricetrin-3-O-rhamnoside (1) against COX-2 was insignificant while that of the other three compounds 2-4 was low. The AChE inhibitory activity of the alkaloid, trans-N-(p-coumaroyl) serotonin was high, with a percentage inhibitory activity of 72.6% and an IC50 value of 15.0 µg/mL. The rest of the compounds only had moderate activity. Croton menyharthii leaf extracts and isolated compounds inhibit α-glucosidase at very low IC50 values compared to the synthetic drug acarbose. Structure activity relationship of the isolated flavonols 1-3 is briefly outlined. Compounds 1-4 and the leaf extracts exhibited a broad spectrum of activities. This validates the

  12. Antimicrobial and Selected In Vitro Enzyme Inhibitory Effects of Leaf Extracts, Flavonols and Indole Alkaloids Isolated from Croton menyharthii

    Directory of Open Access Journals (Sweden)

    Johannes Van Staden

    2013-10-01

    Full Text Available Croton species are used in folk medicine in the management of infections, inflammation and oxidative stress-related diseases. In order to isolate, characterize and evaluate the bioactive constituents of Croton menyharthii Pax leaf extracts, repeated column fractionation of the ethyl acetate fraction from a 20% aqueous methanol crude extract afforded three flavonols identified by NMR (1D and 2D spectroscopic methods as myricetrin-3-O-rhamnoside (myricetrin, 1, quercetin-3-O-rhamnoside (2 and quercetin (3 along with an indole alkaloid, (E-N-(4-hydroxycinnamoyl-5-hydroxytryptamine, [trans-N-(p-coumaroyl serotonin, 4]. All the compounds are reported from the leaf extract of this plant for the first time. The crude extracts, four solvent fractions (hexane, DCM, ethyl acetate and butanol and isolated compounds obtained from the leaves were evaluated for their inhibitory effects on selected bacteria, a fungus (Candida albicans, cyclooxygenase (COX-2, α-glucosidase and acetylcholinesterase (AChE. Amongst the compounds, quercetin (3 was the most active against Bacillus subtilis and Candida albicans while myricetrin-3-O-rhamnoside (1 and trans-N-(p-coumaroyl serotonin (4 were the most active compounds against Escherichia coli, Klebsiella pneumonia and Staphylococcus aureus. The inhibitory activity of myricetrin-3-O-rhamnoside (1 against COX-2 was insignificant while that of the other three compounds 2–4 was low. The AChE inhibitory activity of the alkaloid, trans-N-(p-coumaroyl serotonin was high, with a percentage inhibitory activity of 72.6% and an IC50 value of 15.0 µg/mL. The rest of the compounds only had moderate activity. Croton menyharthii leaf extracts and isolated compounds inhibit α-glucosidase at very low IC50 values compared to the synthetic drug acarbose. Structure activity relationship of the isolated flavonols 1–3 is briefly outlined. Compounds 1–4 and the leaf extracts exhibited a broad spectrum of activities. This validates the

  13. Industrial application of selected lactic acid bacteria isolated from local semolinas for typical sourdough bread production.

    Science.gov (United States)

    Corona, Onofrio; Alfonzo, Antonio; Ventimiglia, Giusi; Nasca, Anna; Francesca, Nicola; Martorana, Alessandra; Moschetti, Giancarlo; Settanni, Luca

    2016-10-01

    strict relation was found among the productions carried out by W. cibaria strains. This study proved the suitability of the selected strains of L. citreum and W. cibaria for industrial-scale level applications in sourdough bread production.

  14. Adjustment of pH of enrichment media might improve selective isolation of MRSA from pig samples

    DEFF Research Database (Denmark)

    Cavaco, Lina; Agersø, Yvonne; Mordhorst, Hanne

    2011-01-01

    concentrations has been the main selection. We hypothesized that also pH adjustment could be used for selection of this species. In this study we compared the growth of MRSA and back ground flora in enrichment media at several different combinations of salt and pH. Background flora isolates were obtained from...... strains was completely inhibited by pH ≤5.5 at any salt concentrations tested (5- 6,5%), whereas the growth of Proteus spp. was only inhibited totally at pH of 4.5, but the growth rate could be reduced combining pH at 5 or 5.5 with high salt concentrations. In conclusion, pH adjustment of enrichment media...... for growth in 12 different combinations of salt (5%- 6.5%) and pH (4.5-5.5). For assessment of the growth of S. aureus in pH adjusted media, further 14 MRSA and 13 MSSA were tested in a similar way. In a preliminary study using reference strains (data not shown) it was observed that pH ≤5.5 as well as salt...

  15. Incarceration of Higher-Order Fullerenes within Cyclotriveratrylene-Based Hemicarcerands Allows Selective Isolation of C76 , C78 , and C84 from a Commercial Fullerene Mixture.

    Science.gov (United States)

    Liu, Kuang-Shun; Li, Ming-Jhe; Lai, Chien-Chen; Chiu, Sheng-Hsien

    2016-11-21

    Size-complementary cyclotriveratrylene (CTV)-based hosts can incarcerate C76 , C78 , and C84 , thus allowing the selective isolation of these higher-order fullerenes from a commercially available mixture of fullerenes. The hemicarceplexes, formed after the encapsulation of the size-complementary fullerenes within the hosts, are isolated by column chromatography and released at elevated temperature, thereby leading to the isolation of C76 /C78 and C84 in good purities (up to 95 and 88 %, respectively). © 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. In vitro selection of meropenem resistance among ceftazidime-avibactam resistant, meropenem susceptible Klebsiella pneumoniae isolates with variant KPC-3 carbapenemases.

    Science.gov (United States)

    Shields, Ryan K; Nguyen, M Hong; Press, Ellen G; Chen, Liang; Kreiswirth, Barry N; Clancy, Cornelius J

    2017-02-27

    Ceftazidime-avibactam resistance is mediated by blaKPC-3 mutations, which restore carbapenem susceptibility. We subjected blaKPC-3 mutant (n=5) and wild-type (n=2) K. pneumoniae isolates to serial meropenem passage. Meropenem MICs against all isolates increased. Ompk36 porin mutations evolved in 5 isolates, including those with wild-type blaKPC-3 In different passage lineages, blaKPC-3 mutations reverted to wild-type, were replaced by new mutations, or were retained. Carbapenem treatment of ceftazidime-avibactam resistant K. pneumoniae infections may select for carbapenem resistance.

  17. Isolated Calyx Mistaken for a Cyst: Inappropriately Performed Catheter-Directed Sclerotherapy and Safe Removal of the Catheter After Selective Embolization

    Energy Technology Data Exchange (ETDEWEB)

    Gwak, Jng Won, E-mail: jjungwonie@hanmail.net; Lee, Seung Hwa, E-mail: gareureung@daum.net; Chung, Hwan Hoon, E-mail: chungmic@korea.ac.kr; Je, Bo Kyung, E-mail: purity21@hanmail.net; Yeom, Suk kyu, E-mail: pagoda20@hanmail.net [Korea University College of Medicine, Department of Radiology, Ansan Hospital (Korea, Republic of); Sung, Deuk Jae, E-mail: urora@korea.ac.kr [Korea University College of Medicine, Department of Radiology, Anam Hospital (Korea, Republic of)

    2015-02-15

    We present a case of isolated calyx that was mistaken for a large cyst. A 47-year-old woman was referred for sclerotherapy of a large cystic lesion on her left kidney. Computed tomography (CT) and ultrasound showed that the cystic lesion was a large cyst. We noticed that the cystic lesion was not a typical simple cyst, even after two sessions of catheter-mediated sclerotherapy. Isolated calyx was presumed by medical history review and was confirmed by aspirated fluid analysis and far delayed-phase CT after intravenous contrast injection. We performed meticulous selective arterial embolization for an isolated calyx and inserted a catheter that could be removed without complication.

  18. Isolation of an Escherichia coli K-12 mutant strain able to form biofilms on inert surfaces: involvement of a new ompR allele that increases curli expression.

    Science.gov (United States)

    Vidal, O; Longin, R; Prigent-Combaret, C; Dorel, C; Hooreman, M; Lejeune, P

    1998-05-01

    Classical laboratory strains of Escherichia coli do not spontaneously colonize inert surfaces. However, when maintained in continuous culture for evolution studies or industrial processes, these strains usually generate adherent mutants which form a thick biofilm, visible with the naked eye, on the wall of the culture apparatus. Such a mutant was isolated to identify the genes and morphological structures involved in biofilm formation in the very well characterized E. coli K-12 context. This mutant acquired the ability to colonize hydrophilic (glass) and hydrophobic (polystyrene) surfaces and to form aggregation clumps. A single point mutation, resulting in the replacement of a leucine by an arginine residue at position 43 in the regulatory protein OmpR, was responsible for this phenotype. Observations by electron microscopy revealed the presence at the surfaces of the mutant bacteria of fibrillar structures looking like the particular fimbriae described by the Olsén group and designated curli (A. Olsén, A. Jonsson, and S. Normark, Nature 338:652-655, 1989). The production of curli (visualized by Congo red binding) and the expression of the csgA gene encoding curlin synthesis (monitored by coupling a reporter gene to its promoter) were significantly increased in the presence of the ompR allele described in this work. Transduction of knockout mutations in either csgA or ompR caused the loss of the adherence properties of several biofilm-forming E. coli strains, including all those which were isolated in this work from the wall of a continuous culture apparatus and two clinical strains isolated from patients with catheter-related infections. These results indicate that curli are morphological structures of major importance for inert surface colonization and biofilm formation and demonstrate that their synthesis is under the control of the EnvZ-OmpR two-component regulatory system.

  19. Hybridization, natural selection, and evolution of reproductive isolation: a 25-years survey of an artificial sympatric area between two mosquito sibling species of the Aedes mariae complex.

    Science.gov (United States)

    Urbanelli, Sandra; Porretta, Daniele; Mastrantonio, Valentina; Bellini, Romeo; Pieraccini, Giuseppe; Romoli, Riccardo; Crasta, Graziano; Nascetti, Giuseppe

    2014-10-01

    Natural selection can act against maladaptive hybridization between co-occurring divergent populations leading to evolution of reproductive isolation among them. A critical unanswered question about this process that provides a basis for the theory of speciation by reinforcement, is whether natural selection can cause hybridization rates to evolve to zero. Here, we investigated this issue in two sibling mosquitoes species, Aedes mariae and Aedes zammitii, that show postmating reproductive isolation (F1 males sterile) and partial premating isolation (different height of mating swarms) that could be reinforced by natural selection against hybridization. In 1986, we created an artificial sympatric area between the two species and sampled about 20,000 individuals over the following 25 years. Between 1986 and 2011, the composition of mating swarms and the hybridization rate between the two species were investigated across time in the sympatric area. Our results showed that A. mariae and A. zammitii have not completed reproductive isolation since their first contact in the artificial sympatric area. We have discussed the relative role of factors such as time of contact, gene flow, strength of natural selection, and biological mechanisms causing prezygotic isolation to explain the observed results. © 2014 The Author(s). Evolution © 2014 The Society for the Study of Evolution.

  20. Nucleotide sequence analysis of a DNA region involved in capsular polysaccharide biosynthesis reveals the molecular basis of the nontypeability of two Actinobacillus pleuropneumoniae isolates.

    Science.gov (United States)

    Ito, Hiroya; Ogawa, Torata; Fukamizu, Dai; Morinaga, Yuiko; Kusumoto, Masahiro

    2016-11-01

    The aim of our study was to reveal the molecular basis of the serologic nontypeability of 2 Actinobacillus pleuropneumoniae field isolates. Nine field strains of A. pleuropneumoniae, the causative agent of porcine pleuropneumonia, were isolated from pigs raised on the same farm and sent to our diagnostic laboratory for serotyping. Seven of the 9 strains were identified as serovar 15 strains by immunodiffusion tests. However, 2 strains, designated FH24-2 and FH24-5, could not be serotyped with antiserum prepared against serovars 1-15. Strain FH24-5 showed positive results in 2 serovar 15-specific PCR tests, whereas strain FH24-2 was only positive in 1 of the 2 PCR tests. The nucleotide sequence analysis of gene clusters involved in capsular polysaccharide biosynthesis of the 2 nontypeable strains revealed that both had been rendered nontypeable by the action of ISApl1, a transposable element of A. pleuropneumoniae belonging to the IS30 family. The results showed that ISApl1 of A. pleuropneumoniae can interfere with both the serologic and molecular typing methods, and that nucleotide sequence analysis across the capsular gene clusters is the best means of determining the cause of serologic nontypeability in A. pleuropneumoniae. © 2016 The Author(s).

  1. Isolation and characterization of Streptomyces, Actinoplanes, and Methylibium strains that are involved in degradation of natural rubber and synthetic poly(cis-1,4-isoprene).

    Science.gov (United States)

    Imai, Shunsuke; Ichikawa, Kazuya; Muramatsu, Yuki; Kasai, Daisuke; Masai, Eiji; Fukuda, Masao

    2011-12-10

    Rubber-degrading bacteria were screened for the production of clearing zones around their colonies on latex overlay agar plates. Novel three bacteria, Streptomyces sp. strain LCIC4, Actinoplanes sp. strain OR16, and Methylibium sp. strain NS21, were isolated. To the best of our knowledge, this is the first report on the isolation of a Gram-negative rubber-degrading bacterium other than γ-proteobacteria. Gel permeation chromatography analysis revealed that these strains degraded poly(cis-1,4-isoprene) to low-molecular-weight products. The occurrence of aldehyde groups in the degradation products by NS21 was suggested by staining with Schiff's reagent and ¹H-nuclear magnetic resonance spectroscopy. The lcp gene of LCIC4, which showed 99% amino acid sequence identity with that of Streptomyces sp. strain K30, was cloned, and contained a putative twin-arginine motif at its N terminus. It is located next to oxiB, which is estimated to be responsible for oxidation of degradation intermediate of rubber in K30. Southern hybridization analysis using LCIC4 lcp probe revealed the presence of a lcp-homolog in OR16. These results suggest that the lcp-homologs are involved in rubber degradation in LCIC4 and OR16.

  2. Brain-derived neurotrophic factor controls functional differentiation and microcircuit formation of selectively isolated fast-spiking GABAergic interneurons.

    Science.gov (United States)

    Berghuis, Paul; Dobszay, Marton B; Sousa, Kyle M; Schulte, Gunnar; Mager, Peter P; Härtig, Wolfgang; Görcs, Tamás J; Zilberter, Yuri; Ernfors, Patrik; Harkany, Tibor

    2004-09-01

    GABAergic interneurons with high-frequency firing, fast-spiking (FS) cells, form synapses on perisomatic regions of principal cells in the neocortex and hippocampus to control the excitability of cortical networks. Brain-derived neurotrophic factor (BDNF) is essential for the differentiation of multiple interneuron subtypes and the formation of their synaptic contacts. Here, we examined whether BDNF, alone or in conjunction with sustained KCl-induced depolarization, drives functional FS cell differentiation and the formation of inhibitory microcircuits. Homogeneous FS cell cultures were established by target-specific isolation using the voltage-gated potassium channel 3.1b subunit as the selection marker. Isolated FS cells expressed parvalbumin, were surrounded by perineuronal nets, formed immature inhibitory connections and generated slow action potentials at 12 days in vitro. Brain-derived neurotrophic factor (BDNF) promoted FS cell differentiation by increasing the somatic diameter, dendritic branching and the frequency of action potential firing. In addition, BDNF treatment led to a significant up-regulation of synaptophysin and vesicular GABA transporter expression, components of the synaptic machinery critical for GABA release, which was paralleled by an increase in synaptic strength. Long-term membrane depolarization alone was detrimental to dendritic branching. However, we observed that BDNF and KCl exerted additive effects, as reflected by the significantly accelerated maturation of synaptic contacts and high discharge frequencies, and was required for the formation of reciprocal connections between FS cells. Our results show that BDNF, along with membrane depolarization, is critical for FS cells to establish inhibitory circuitries during corticogenesis.

  3. Screening and selection of most potent diazotrophic cyanobacterial isolate exhibiting natural tolerance to rice field herbicides for exploitation as biofertilizer.

    Science.gov (United States)

    Singh, Surendra; Datta, Pallavi

    2006-01-01

    Periodic applications of heavy dosages of herbicides in modern rice-agriculture are a necessary evil for obtaining high crop productivity. Such herbicides are not only detrimental to weeds but biofertilizer strains of diazotrophic cyanobacteria also. It is therefore, essential to screen and select such biofertilizer strains of diazotrophic cyanobacteria exhibiting natural tolerance to common rice-field herbicides that can be further improved by mutational techniques to make biofertilizer technology a viable one. Therefore, efforts have been made to screen five dominant diazotrophic cyanobacterial forms e.g. filamentous heterocystous Nostoc punctiforme , Nostoc calcicola , Anabaena variabilis and unicellular Gloeocapsa sp. and Aphanocapsa sp. along with standard laboratory strain Nostoc muscorum ISU against increasing concentrations (0-100 mg l(-1) of four commercial grade common rice-field herbicides i.e. Arozin, Butachlor, Alachlor and 2,4-D under diazotrophic growth conditions. The lethal and IGC(50) concentrations for all four herbicides tested were found highest for A. variabilis as compared to other test cyanobacteria. The lowest reduction in chlorophyll a content, photosynthetic oxygen evolution, and N(2)-fixation was found in A. variabilis as compared to other rice field isolates and standard laboratory strain N. muscorum ISU. On the basis of prolong survival potential and lowest reductions in vital metabolic activities tested at IGC(50) concentration of four herbicides, it is concluded that A. variabilis is the most potent and promising cyanobacterial isolate as compared with other forms. This could be further improved by mutational techniques for exploitation as most potential and viable biofertilizer strain.

  4. Chapter 4: considerations during the site selection. Public involvement and acceptance; Capitulo 4: consideracoes durante a selecao de sitio. Envolvimento e aceitacao publicos

    Energy Technology Data Exchange (ETDEWEB)

    Atala, Drausio Lima

    2009-07-01

    The public involvement and acceptance is to be considered a necessary and desirable part of the process of the nuclear site selection and allows to the entrepreneur to consult and incorporate the considerations of the interested people in the decision process.

  5. Isolation, selection and evaluation of yeasts for use in fermentation of coffee beans by the wet process.

    Science.gov (United States)

    de Melo Pereira, Gilberto Vinícius; Soccol, Vanete Thomaz; Pandey, Ashok; Medeiros, Adriane Bianchi Pedroni; Andrade Lara, João Marcos Rodrigues; Gollo, André Luiz; Soccol, Carlos Ricardo

    2014-10-01

    During wet processing of coffee, the ripe cherries are pulped, then fermented and dried. This study reports an experimental approach for target identification and selection of indigenous coffee yeasts and their potential use as starter cultures during the fermentation step of wet processing. A total of 144 yeast isolates originating from spontaneously fermenting coffee beans were identified by molecular approaches and screened for their capacity to grow under coffee-associated stress conditions. According to ITS-rRNA gene sequencing, Pichia fermentans and Pichia kluyveri were the most frequent isolates, followed by Candida Candida glabrata, quercitrusa, Saccharomyces sp., Pichia guilliermondii, Pichia caribbica and Hanseniaspora opuntiae. Nine stress-tolerant yeast strains were evaluated for their ability to produce aromatic compounds in a coffee pulp simulation medium and for their pectinolytic activity. P. fermentans YC5.2 produced the highest concentrations of flavor-active ester compounds (viz., ethyl acetate and isoamyl acetate), while Saccharomyces sp. YC9.15 was the best pectinase-producing strain. The potential impact of these selected yeast strains to promote flavor development in coffee beverages was investigated for inoculating coffee beans during wet fermentation trials at laboratory scale. Inoculation of a single culture of P. fermentans YC5.2 and co-culture of P. fermentans YC5.2 and Saccharomyces sp. YC9.15 enhanced significantly the formation of volatile aroma compounds during the fermentation process compared to un-inoculated control. The sensory analysis indicated that the flavor of coffee beverages was influenced by the starter cultures, being rated as having the higher sensory scores for fruity, buttery and fermented aroma. This demonstrates a complementary role of yeasts associated with coffee quality through the synthesis of yeast-specific volatile constituents. The yeast strains P. fermentans YC5.2 and Saccharomyces sp. YC9.15 have a great

  6. A Best Practice for Selecting Your QEP: Techniques for Facilitating Mission-Based Stakeholder Involvement in the Selection of the Institutional Quality Enhancement Plan

    Science.gov (United States)

    Tincher-Ladner, Lynn

    2009-01-01

    How can comprehensive community colleges serve students with diverse learning needs today while determining a single Quality Enhancement Plan (QEP) to improve student learning? During the reaffirmation planning process, Mississippi Gulf Coast Community College (MGCCC) recognized this challenge and responded by designing a plan for the selection of…

  7. Balancing selection and genetic drift at major histocompatibility complex class II genes in isolated populations of golden snub-nosed monkey (Rhinopithecus roxellana

    Directory of Open Access Journals (Sweden)

    Luo Mao-Fang

    2012-10-01

    Full Text Available Abstract Background Small, isolated populations often experience loss of genetic variation due to random genetic drift. Unlike neutral or nearly neutral markers (such as mitochondrial genes or microsatellites, major histocompatibility complex (MHC genes in these populations may retain high levels of polymorphism due to balancing selection. The relative roles of balancing selection and genetic drift in either small isolated or bottlenecked populations remain controversial. In this study, we examined the mechanisms maintaining polymorphisms of MHC genes in small isolated populations of the endangered golden snub-nosed monkey (Rhinopithecus roxellana by comparing genetic variation found in MHC and microsatellite loci. There are few studies of this kind conducted on highly endangered primate species. Results Two MHC genes were sequenced and sixteen microsatellite loci were genotyped from samples representing three isolated populations. We isolated nine DQA1 alleles and sixteen DQB1 alleles and validated expression of the alleles. Lowest genetic variation for both MHC and microsatellites was found in the Shennongjia (SNJ population. Historical balancing selection was revealed at both the DQA1 and DQB1 loci, as revealed by excess non-synonymous substitutions at antigen binding sites (ABS and maximum-likelihood-based random-site models. Patterns of microsatellite variation revealed population structure. FST outlier analysis showed that population differentiation at the two MHC loci was similar to the microsatellite loci. Conclusions MHC genes and microsatellite loci showed the same allelic richness pattern with the lowest genetic variation occurring in SNJ, suggesting that genetic drift played a prominent role in these isolated populations. As MHC genes are subject to selective pressures, the maintenance of genetic variation is of particular interest in small, long-isolated populations. The results of this study may contribute to captive breeding and

  8. Selection of Beauveria bassiana sensu lato and Metarhizium anisopliae sensu lato isolates as microbial control agents against the boll weevil (Anthonomus grandis) in Argentina.

    Science.gov (United States)

    Nussenbaum, A L; Lecuona, R E

    2012-05-01

    The boll weevil (Anthonomus grandis) is the main pest of cotton in the Americas. The aim of this work was to evaluate isolates of the entomopathogenic fungi Beauveria bassiana sensu lato and Metarhizium anisopliae sensu lato virulent against A. grandis. Screening was performed to evaluate the pathogenicity of 28 isolates of M. anisopliae s.l. and 66 isolates of B. bassiana s.l. against boll weevil adults. To select the isolates, LC(50) values of the most virulent isolates were calculated, and compatibility between the fungi and insecticides was studied. In addition, the effects of these isolates on the feeding behavior of the adults were evaluated. Isolates Ma 50 and Ma 20 were the most virulent against A. grandis and their LC(50) values were 1.13×10(7) and 1.20×10(7) conidia/ml, respectively. In addition, these isolates were compatible with pyrethroid insecticides, but none with endosulfan. On the other hand, infected females reduced the damage caused by feeding on the cotton squares and their weight gain. This shows that entomopathogenic fungi cause mortality in the insects, but also these fungi could influence the feeding behavior of the females. In summary, these results indicate the possibility of the use of M. anisopliae s.l. as a microbiological control agent against boll weevils. Also, this species could be included in an Integrated Pest Management program.

  9. Thymic commitment of regulatory T cells is a pathway of TCR-dependent selection that isolates repertoires undergoing positive or negative selection.

    Science.gov (United States)

    Coutinho, A; Caramalho, I; Seixas, E; Demengeot, J

    2005-01-01

    The seminal work of Le Douarin and colleagues (Ohki et al. 1987; Ohki et al. 1988; Salaun et al. 1990; Coutinho et al. 1993) first demonstrated that peripheral tissue-specific tolerance is centrally established in the thymus, by epithelial stromal cells (TEC). Subsequent experiments have shown that TEC-tolerance is dominant and mediated by CD4 regulatory T cells (Treg) that are generated intrathymically by recognition of antigens expressed on TECs (Modigliani et al. 1995; Modigliani et al. 1996a). From these and other observations, in 1996 Modigliani and colleagues derived a general model for the establishment and maintenance of natural tolerance (MM96) (Modigliani et al. 1996b), with two central propositions: (1) T cell receptor (TCR)-dependent sorting of emergent repertoires generates TEC-specific Treg displaying the highest TCR self-affinities below deletion thresholds, thus isolating repertoires undergoing positive and negative selection; (2) Treg are intrathymically committed (and activated) for a unique differentiative pathway with regulatory effector functions. The model explained the embryonic/perinatal time window of natural tolerance acquisition, by developmental programs determining (1) TCR multireactivity, (2) the cellular composition in the thymic stroma (relative abundance of epithelial vs hemopoietic cells), and (3) the dynamics of peripheral lymphocyte pools, built by accumulation of recent thymic emigrants (RTE) that remain recruitable to regulatory functions. We discuss here the MM96 in the light of recent results demonstrating the promiscuous expression of tissue-specific antigens by medullary TECs (Derbinski et al. 2001; Anderson et al. 2002; Gotter et al. 2004) and indicating that Treg represent a unique differentiative pathway (Fontenot et al. 2003; Hori et al. 2003; Khattri et al. 2003), which is adopted by CD4 T cells with high avidity for TEC-antigens (Bensinger et al. 2001; Jordan et al. 2001; Apostolou et al. 2002). In the likelihood that

  10. Isolation of Two Unknown Genes Potentially Involved in Differentiation of the Hematopoietic Pathway, and Studies of Spermidine/Spermine Acetyltransferase Regulation

    Energy Technology Data Exchange (ETDEWEB)

    Kubera, C.; Gavin, I.; Huberman, E.

    2002-01-01

    Differential display identified a number of candidate genes involved with growth and differentiation in the human leukemia cell lines HL-60 and HL-525. Two of these genes were previously unknown, and one is the gene for the enzyme spermidine/spermine acetyltransferase (SSAT). One of our objectives is to isolate and sequence the unknown genes, 631A1 and 510C1, in order to characterize them and determine their functions. The other is to determine how SSAT is regulated, and look at how the polyamines that SSAT regulates effect macrophage differentiation. By screening the CEM T-cell DNA library and the fetal brain library, we were able to identify clones that had inserts with homology to the 631A1 cDNA probe sequence. The insert was amplified using the polymerase chain reaction (PCR) and is currently being sent to the University of Chicago for automated sequencing. The library screens for 510C1 are currently underway, but hybridization of the 510C1 cDNA probe with nylon membranes containing CEM library phage DNA produced strong signal, indicating the gene is there. SSAT experiments identified that the rate-limiting enzyme that marks the polyamines spermidine and spermine for degradation is regulated by PKC and a transcription factor called Nrf2. The knowledge of regulation and function of these genes involved in macrophage differentiation will provide new insight into this cellular process, potentially making it possible to discover the roots of the problems that cause cancerous diseases.

  11. Isolation and identification of novel genes involved in artemisinin production from flowers of Artemisia annua using suppression subtractive hybridization and metabolite analysis.

    Science.gov (United States)

    Liu, Shuoqian; Tian, Na; Li, Juan; Huang, Jianan; Liu, Zhonghua

    2009-11-01

    Malaria is a global health problem that threatens 300-500 million people and kills more than one million people annually. Artemisinin is highly effective against multidrug-resistant Plasmodium falciparum and it has been widely used as part of the artemisinin-based combination therapies against malaria. To elucidate the biosynthetic pathway of artemisinin and to clone related genes in Artemisia annua, differentially expressed genes between blooming flowers and flower buds were isolated and characterized by a combined approach of suppression subtractive hybridization (SSH) and metabolite analysis. A total of 350 cDNA clones from a subtractive cDNA library were randomly picked, sequenced and analyzed and 253 high-quality sequences were obtained. BLASTX comparisons indicated that about 9.9 % of the clones encoded enzymes involved in isoprenoid (including artemisinin) biosynthesis. The expression of 4 gene transcripts involved in artemisinin biosynthesis was examined by RT-PCR and the results confirmed the higher expression of these transcripts in blooming flowers than in flower buds. In addition, 2 putative transcript factors transparenta testa glabra 1 (TTG1) and ENHANCER OF GLABRA3 (GL3), which promote trichome initiation, were presented in the library. Finally, this study demonstrated that the increase of expression level of the putative TTG1 gene correlated with the improvement of glandular trichome density and artemisinin production in A. annua leaves. The subtractive cDNA library described in the present study provides important candidate genes for future research in order to increase the artemisinin content in A. annua.

  12. Comparative study of antifungal activities of six selected essential oils against fungal isolates from cheese wagashi in Benin.

    Science.gov (United States)

    Sessou, P; Farougou, S; Ahounou, S; Hounnankpon, Y; Azokpota, P; Youssao, I; Sohounhloue, D

    2013-12-01

    The study has compared the antifungal efficacy of six essential oils, Cinnamomum zeylanicum, Cymbopogon citratus, Ocimum gratissimum, Pimenta racemosa, Syzygium aromaticum and Zingiber officinale, tested in culture medium and in traditional cheese wagashi system against moulds belonging to Aspergillus, Penicillium, Fusarium and Scopulariopsis genera in perspective to select the most actives as substitutes of chemical preservatives for wagashi preservation. Results obtained from this work indicated that Syzygium aromaticum, Pimenta racemosa, Ocimum gratissimum and Cymbopogon citratus essentials oils were the most actives extracts at in vitro assay in decreasing order with strong fungistatic activity against the isolates tested; the pronounced activity was provided by S. aromaticum essential oil. The effectiveness of these actives oils on the less sensitive moulds common to these oils showed that, among these extracts that of Syzygium aromaticum in particular exerted high sporale reduction against all the strains tested. In sum, Syzygium aromaticum essential oil possessed the highest antifungal activity both in culture medium and in wagashi system. Essential oils of C. citratus, O. gratissimum, P. racemosa and above all that of S. aromaticum, among the six extracts investigated, were the most promising oils as wagashi additives in substitution of synthetic chemicals ones to extend shelf life time of this by-product of milk for its valorization. Further studies are needed to be performed on the safety of oils for human, the shelf life time of this cheese and its acceptability when treated with essential oils to reduce and control pathogen contamination or native microflora.

  13. Antibacterial Effects of Origanum vulgare Essence Against Multidrug-Resistant Acinetobacter baumannii Isolated From Selected Hospitals of Tehran, Iran

    Directory of Open Access Journals (Sweden)

    Saghi

    2015-02-01

    Full Text Available Background Infection due to Acinetobacter baumannii has become a significant challenge to modern healthcare systems. The rapid emergence and global dissemination of A. baumannii as a major nosocomial pathogen is remarkable and it demonstrates its successful adaptation to the 21st century hospital environment. Recent studies have discussed about essential oil of Origanum vulgare against a range of bacteria, including various species of Staphylococcus, Pseudomonas, Bacillus and Escherichia coli. Objectives The present study aimed to investigate the inhibitory effects O. vulgare essence against multidrug-resistant (MDR strains of A. baumannii from selected hospitals in Tehran, Iran. Materials and Methods This oil was obtained using the hydrodistillation method and analyzed by gas chromatography mass spectrography (GC/MS. The antimicrobial activity against MDR isolates was achieved using disc diffusion method and macro-broth dilution assay. Results Analysis of the essential oil revealed the presence of pulegone (68.59% piperitone (7.8%, piperitenone (7.8%, 1, 8-cineole (1.3%, and carvacrol (1.6% as the major components. The results showed a significant activity against MDR A. baumannii with inhibition zones and minimal inhibitory concentration values in the ranges of 7-15 mm and 20-35 µL/mL respectively. Conclusions This investigation showed that the essence oil of O. vulgare had a potent antimicrobial activity against MDR A. baumannii. Further research is required to evaluate the practical values of therapeutic applications.

  14. Isolation of the copper redox steps in the standard selective catalytic reduction on Cu-SSZ-13.

    Science.gov (United States)

    Paolucci, Christopher; Verma, Anuj A; Bates, Shane A; Kispersky, Vincent F; Miller, Jeffrey T; Gounder, Rajamani; Delgass, W Nicholas; Ribeiro, Fabio H; Schneider, William F

    2014-10-27

    Operando X-ray absorption experiments and density functional theory (DFT) calculations are reported that elucidate the role of copper redox chemistry in the selective catalytic reduction (SCR) of NO over Cu-exchanged SSZ-13. Catalysts prepared to contain only isolated, exchanged Cu(II) ions evidence both Cu(II) and Cu(I) ions under standard SCR conditions at 473 K. Reactant cutoff experiments show that NO and NH3 together are necessary for Cu(II) reduction to Cu(I). DFT calculations show that NO-assisted NH3 dissociation is both energetically favorable and accounts for the observed Cu(II) reduction. The calculations predict in situ generation of Brønsted sites proximal to Cu(I) upon reduction, which we quantify in separate titration experiments. Both NO and O2 are necessary for oxidation of Cu(I) to Cu(II), which DFT suggests to occur by a NO2 intermediate. Reaction of Cu-bound NO2 with proximal NH4(+) completes the catalytic cycle. N2 is produced in both reduction and oxidation half-cycles.

  15. Isolation of the Copper Redox Steps in the Standard Selective Catalytic Reduction on Cu-SSZ-13

    Energy Technology Data Exchange (ETDEWEB)

    Paolucci, Christopher; Verma, Anuj A.; Bates, Shane A.; Kispersky, Vincent F.; Miller, Jeffrey T.; Gounder, Rajmani; Delgass, Nick; Ribeiro, Fabio; Schneider, William F.

    2014-10-27

    Operando X-ray absorption experiments and density functional theory (DFT) calculations are reported that elucidate the role of copper redox chemistry in the selective catalytic reduction (SCR) of NO over Cu-exchanged SSZ-13. Catalysts prepared to contain only isolated, exchanged CuII ions evidence both CuII and CuI ions under standard SCR conditions at 473 K. Reactant cutoff experiments show that NO and NH3 together are necessary for CuII reduction to CuI. DFT calculations show that NO-assisted NH3 dissociation is both energetically favorable and accounts for the observed CuII reduction. The calculations predict in situ generation of Brønsted sites proximal to CuI upon reduction, which we quantify in separate titration experiments. Both NO and O2 are necessary for oxidation of CuI to CuII, which DFT suggests to occur by a NO2 intermediate. Reaction of Cu-bound NO2 with proximal NH4 + completes the catalytic cycle. N2 is produced in both reduction and oxidation half-cycles.

  16. Selective inhibitory effects of niflumic acid on 5-HT-induced contraction of the rat isolated stomach fundus.

    Science.gov (United States)

    Scarparo, H C; Santos, G C; Leal-Cardoso, J H; Criddle, D N

    2000-06-01

    The effects of niflumic acid (NFA), an inhibitor of calcium-activated chloride currents I(Cl(Ca)), were compared with the actions of the voltage-dependent calcium channel (VDCC) blocker nifedipine on 5-hydroxtryptamine (5-HT)- and acetylcholine (ACh)-induced contractions of the rat isolated fundus. NFA (1 - 30 microM) elicited a concentration-dependent inhibition of contractions induced by 5-HT (10 microM) with a reduction to 15. 5+/-6.0% of the control value at 30 microM. 1 microM nifedipine reduced 5-HT-induced contraction to 15.2+/-4.9% of the control, an effect not greater in the additional presence of 30 microM NFA. In contrast, the contractile response to ACh (10 microM) was not inhibited by NFA in concentrations /=10 microM. Our results show that NFA can exert selective inhibitory effects on the chloride-dependent 5-HT-induced contractions of the rat fundus. The data support the hypothesis that activation of Cl((Ca)) channels leading to calcium entry via VDCCs is a mechanism utilized by 5-HT, but not by ACh, to elicit contraction of the rat fundus.

  17. Actinobacteria phylogenomics, selective isolation from an iron oligotrophic environment and siderophore functional characterization, unveil new desferrioxamine traits.

    Science.gov (United States)

    Cruz-Morales, Pablo; Ramos-Aboites, Hilda E; Licona-Cassani, Cuauhtémoc; Selem-Mójica, Nelly; Mejía-Ponce, Paulina M; Souza-Saldívar, Valeria; Barona-Gómez, Francisco

    2017-09-01

    Desferrioxamines are hydroxamate siderophores widely conserved in both aquatic and soil-dwelling Actinobacteria. While the genetic and enzymatic bases of siderophore biosynthesis and their transport in model families of this phylum are well understood, evolutionary studies are lacking. Here, we perform a comprehensive desferrioxamine-centric (des genes) phylogenomic analysis, which includes the genomes of six novel strains isolated from an iron and phosphorous depleted oasis in the Chihuahuan desert of Mexico. Our analyses reveal previously unnoticed desferrioxamine evolutionary patterns, involving both biosynthetic and transport genes, likely to be related to desferrioxamines chemical diversity. The identified patterns were used to postulate experimentally testable hypotheses after phenotypic characterization, including profiling of siderophores production and growth stimulation of co-cultures under iron deficiency. Based in our results, we propose a novel des gene, which we term desG, as responsible for incorporation of phenylacetyl moieties during biosynthesis of previously reported arylated desferrioxamines. Moreover, a genomic-based classification of the siderophore-binding proteins responsible for specific and generalist siderophore assimilation is postulated. This report provides a much-needed evolutionary framework, with specific insights supported by experimental data, to direct the future ecological and functional analysis of desferrioxamines in the environment. © FEMS 2017.

  18. Modulation of cytokine gene expression by selected Lactobacillus isolates in the ileum, caecal tonsils and spleen of Salmonella-challenged broilers.

    Science.gov (United States)

    Hu, Jie-Lun; Yu, Hai; Kulkarni, Raveendra R; Sharif, Shayan; Cui, Steve W; Xie, Ming-Yong; Nie, Shao-Ping; Gong, Joshua

    2015-01-01

    Probiotics have been used to control Salmonella colonization in the chicken intestine. Recently, we demonstrated that certain selected Lactobacillus isolates were able to reduce Salmonella infection in the chicken spleen and liver as well as down-regulated Salmonella pathogenicity island 1 virulence gene expression in the chicken caecum. To further understand the mechanisms through which Lactobacillus protected chickens from Salmonella infection, the present study has investigated the Lactobacillus isolate(s)-induced host immune response of chickens to Salmonella enterica serovar Typhimurium infection. A thorough examination of cytokine gene expression in the ileum, caecal tonsils, and spleen on days 1 and 3 post-Salmonella infection showed a dynamic spatial and temporal response to Salmonella infection and Lactobacillus treatments. In most instances, it was evident that treatment of chickens with Lactobacillus isolates could significantly attenuate Salmonella-induced changes in the gene expression profile. These included the genes encoding pro-inflammatory cytokines [lipopolysaccharide-induced TNF factor, interleukin (IL)-6, and IL-8], T helper 1 cytokines [IL-12 and interferon (IFN)-γ], and T helper 2 cytokines (IL-4 and IL-10). Another important observation from the present investigation was that the response induced by a combination of Lactobacillus isolates was generally more effective than that induced by a single Lactobacillus isolate. Our results show that administration of certain selected Lactobacillus isolates can effectively modulate Salmonella-induced cytokine gene expression, and thus help reduce Salmonella infection in chickens.

  19. Different Patterns of pfcrt and pfmdr1 Polymorphisms in P. falciparum Isolates from Nigeria and Brazil: The Potential Role of Antimalarial Drug Selection Pressure

    Science.gov (United States)

    Gbotosho, Grace O.; Folarin, Onikepe A.; Bustamante, Carolina; Pereira da Silva, Luis Hildebrando; Mesquita, Elieth; Sowunmi, Akintunde; Zalis, Mariano G.; Oduola, Ayoade M. J.; Happi, Christian T.

    2012-01-01

    The effect of antimalarial drug selection on pfcrt and pfmdr1 polymorphisms in Plasmodium falciparum isolates from two distinct geographical locations was determined in 70 and 18 P. falciparum isolates from Nigeria and Brazil, respectively, using nested polymerase chain reaction and direct DNA sequencing approaches. All isolates from Brazil and 72% from Nigeria harbored the mutant SVMNT and CVIET pfcrt haplotype, respectively. The pfcrt CVMNT haplotype was also observed in (7%) of the Nigerian samples. One hundred percent (100%) and 54% of the parasites from Brazil and Nigeria, respectively, harbored wild-type pfmdr1Asn86. We provide first evidence of emergence of the CVMNT haplotype in West Africa. The high prevalence of pfcrt CVIET and SVMNT haplotypes in Nigeria and Brazil, respectively, is indicative of different selective pressure by chloroquine and amodiaquine. Continuous monitoring of pfcrt SVMNT haplotype is required in endemic areas of Africa, where artesunate-amodiaquine combination is used for treatment of acute uncomplicated malaria. PMID:22302850

  20. Teachers' Perspective towards Their Involvement in Selection and Organization of Learning Experiences and Implementation of Secondary School Curriculum in Kenya

    Science.gov (United States)

    Kobiah, Lydia Kanake

    2016-01-01

    Teacher empowerment has been the subject of considerable educational research in recent years, but the capacity of teachers in curriculum development especially in selection and organization of learning experiences has received limited empirical attention. The purpose of this study was to establish teachers' perspective towards their involvement…

  1. Selection, assessment of virulence to Alphitobius diaperinus, and Pr1 enzyme production of Beauveria bassiana (Bals. Vuill. isolates cultured at stress temperatures

    Directory of Open Access Journals (Sweden)

    Kelly Christiane Constanski

    2015-12-01

    Full Text Available The entomopathogenic fungus Beauveria bassiana is a promising agent for use in insect control. Its pathogenic activity, as well as other factors such as temperature that can interfere with its development, should be assessed, thus, establishing the foundations for B. bassiana use in biological control programs. The objective of this study was to select and induce tolerance of B. bassiana isolates to high and low temperatures and to assess their virulence before and after exposure to those temperatures. A pre-selection test was performed, in which the tolerance of isolates to stress temperatures was tested and compared to the ideal growth temperature of 25 °C for this organism. For the isolates/temperature combinations resulting in growth, conidia germination and colony-forming units (CFUs were assessed. The isolates Unioeste 4 and Unioeste 40 exhibited >95% germinated conidia at 16 and 31 °C. Thereafter, they underwent four consecutive passages at maximum and minimum tolerated temperatures (10 and 37 °C. A significant difference in germination was observed between the two isolates at all temperatures tested. More CFUs were observed for Unioeste 4 compared to Unioeste 40 at all temperatures, and in the case of the latter, there was no difference in CFU formation at 10 and 25 °C. For both isolates, decreased vegetative growth was observed at 37 °C. Recovery of virulence was observed in both isolates, as determined by insect mortality. No relationship was observed between production of the enzyme Pr1 and the virulence of the isolates.

  2. A hidden Markov model to identify and adjust for selection bias: an example involving mixed migration strategies

    OpenAIRE

    Fieberg, John R.; Paul B Conn

    2014-01-01

    An important assumption in observational studies is that sampled individuals are representative of some larger study population. Yet, this assumption is often unrealistic. Notable examples include online public-opinion polls, publication biases associated with statistically significant results, and in ecology, telemetry studies with significant habitat-induced probabilities of missed locations. This problem can be overcome by modeling selection probabilities simultaneously with other predicto...

  3. Brain regions involved in processing facial identity and expression are differentially selective for surface and edge information.

    Science.gov (United States)

    Harris, Richard J; Young, Andrew W; Andrews, Timothy J

    2014-08-15

    Although different brain regions are widely considered to be involved in the recognition of facial identity and expression, it remains unclear how these regions process different properties of the visual image. Here, we ask how surface-based reflectance information and edge-based shape cues contribute to the perception and neural representation of facial identity and expression. Contrast-reversal was used to generate images in which normal contrast relationships across the surface of the image were disrupted, but edge information was preserved. In a behavioural experiment, contrast-reversal significantly attenuated judgements of facial identity, but only had a marginal effect on judgements of expression. An fMR-adaptation paradigm was then used to ask how brain regions involved in the processing of identity and expression responded to blocks comprising all normal, all contrast-reversed, or a mixture of normal and contrast-reversed faces. Adaptation in the posterior superior temporal sulcus--a region directly linked with processing facial expression--was relatively unaffected by mixing normal with contrast-reversed faces. In contrast, the response of the fusiform face area--a region linked with processing facial identity--was significantly affected by contrast-reversal. These results offer a new perspective on the reasons underlying the neural segregation of facial identity and expression in which brain regions involved in processing invariant aspects of faces, such as identity, are very sensitive to surface-based cues, whereas regions involved in processing changes in faces, such as expression, are relatively dependent on edge-based cues.

  4. Light-triggered selective nitration of PsbO1 in isolated Arabidopsis thylakoid membranes is inhibited by photosynthetic electron transport inhibitors.

    Science.gov (United States)

    Takahashi, Misa; Shigeto, Jun; Sakamoto, Atsushi; Morikawa, Hiromichi

    2016-12-01

    PsbO1 is exclusively nitrated when isolated thylakoid membranes are incubated in a buffer bubbled with nitrogen dioxide (NO2) containing NO2 and nitrite. NO2 is the primary intermediate for this selective nitration. Isolated thylakoid membranes were incubated in NO2-bubbled buffer at 25°C in the light or dark. Protein analysis confirmed the selective nitration of PsbO1. Illumination was found to be essential in PsbO1 nitration. A nitration mechanism whereby nitratable tyrosine residues of PsbO1 are, prior to nitration, selectively photo-oxidized by photosynthetic electron transport to tyrosyl radicals to combine with NO2 to form 3-nitrotyrosine was hypothesized. We tested the electron transport inhibitors 3-(3,4-dichlorophenyl)-1,1- dimethylurea, sodium azide, and 1,5-diphenylcarbazide and found distinct inhibition of nitration of PsbO1. We also propose a possible nitration mechanism.

  5. Genetic diversity of infectious hematopoietic necrosis virus from Feather River and Lake Oroville, California, and virulence of selected isolates for Chinook salmon and rainbow trout.

    Science.gov (United States)

    Bendorf, Christin M; Kelley, Garry O; Yun, Susan C; Kurath, Gael; Andree, Karl B; Hedrick, Ronald P

    2007-12-01

    Infectious hematopoietic necrosis virus (IHNV) is a significant pathogen of young salmonid fishes worldwide but particularly within the historical range of the Pacific Northwest and California. In the Sacramento and San Joaquin River drainages of California, IHNV outbreaks in juvenile Chinook salmon Oncorhynchus tshawytscha have been observed regularly at large production hatcheries, including Coleman National Fish Hatchery (established in 1941) and Feather River State Fish Hatchery (FRH; established in 1967), since facility operations began. Recent severe epidemics at the FRH in 1998 and 2000-2002 prompted investigations into the characteristics and potential sources of virus at this facility. Both phylogenetic analyses of a variable portion of the glycoprotein gene and serologic comparisons based on neutralization with three polyclonal rabbit sera were used to characterize 82 IHNV isolates from the Feather River watershed between 1969 and 2004. All isolates examined were in the L genogroup and belonged to one of three serologic groups typical of IHNV from California. The IHNV isolates from the Feather River area demonstrated a maximum nucleotide sequence divergence of 4.0%, and new isolates appeared to emerge from previous isolates rather than by the introduction of more diverse subgroups from exogenous sources. The earliest isolates examined from the watershed formed the subgroup LI, which disappeared coincidently with a temporal shift to new genetic and serologic types of the larger subgroup LII. Experimental challenges demonstrated no significant differences in the virulence for juvenile Chinook salmon and rainbow trout O. mykiss from selected isolates representing the principal types of IHNV found historically and from recent epidemics at FRH. While most isolates were equally virulent for both host species, one isolate was found to be more virulent for Chinook salmon than for rainbow trout.

  6. Isolation, screening, characterization, and selection of superior rhizobacterial strains as bioinoculants for seedling emergence and growth promotion of Mandarin orange (Citrus reticulata Blanco).

    Science.gov (United States)

    Thokchom, Elizabeth; Kalita, Mohan Chandra; Talukdar, Narayan Chandra

    2014-02-01

    Mandarin orange (MO) is an important fruit crop of tropical and subtropical regions of the world. A total of 217 morphologically distinct rhizobacteria from MO orchards in 3 states of northeastern India were isolated and analyzed for 4 plant-growth-promoting (PGP) attributes: nitrogen fixation, production of indole acetic acid like substances, solubilization of phosphate, and ability to antagonize pathogenic fungi. Isolates were ranked based on in-vitro-assayed PGP attributes, and 10 superior isolates were selected to test their effect on seedling emergence and seedling growth in a completely randomized pot experiment. These 10 isolates increased seedling emergence over a noninoculated control within 45 days after sowing. Five isolates, namely RCE1, RCE2, RCE3, RCE5, and RCE7, significantly increased shoot length, shoot dry biomass, and root dry biomass of 120-day-old seedlings over the noninoculated control. The beneficial effects of 4 selected strains, namely Enterobacter hormaechei RCE-1, Enterobacter asburiae RCE-2, Enterobacter ludwigii RCE-5, and Klebsiella pneumoniae RCE-7, on growth of the seedlings were visible up to 1 year after their transfer to 8 kg capacity pots. These strains were superior both in terms of in-vitro-assayed PGP attributes and of their beneficial effect in low phosphorus soil and, thus, may be promising bioinoculants for promoting early emergence and growth of MO seedlings.

  7. Antidepressant-Like Effects of Sanggenon G, Isolated from the Root Bark of Morus alba, in Rats: Involvement of the Serotonergic System.

    Science.gov (United States)

    Lim, Dong Wook; Jung, Jae-Woo; Park, Ji-Hae; Baek, Nam-In; Kim, Yun Tai; Kim, In-Ho; Han, Daeseok

    2015-01-01

    The root bark of Morus alba is commonly used as an alternative medicine due to its numerous health benefits in humans. However, the antidepressant effects of various active components from M. alba have not been fully elucidated. In this study, we aimed to determine whether sanggenon G, an active compound isolated from the root bark of M. alba, exhibited antidepressant-like activity in rats subjected to forced swim test (FST)-induced depression. Acute treatment of rats with sanggenon G (30 mg/kg, intraperitoneally (i.p.)) significantly reduced immobility time and increased swimming time without any significant change in climbing. Rats treated with sanggenon G also exhibited a decrease in the limbic hypothalamic-pituitary-adrenal (HPA) axis response to the FST, as indicated by attenuation of the corticosterone response and decreased c-Fos immunoreactivity in the hypothalamic paraventricular nucleus (PVN). In addition, the antidepressant-like effects of sanggenon G were significantly inhibited by WAY100635 (1 mg/kg, i.p.; a selective 5-hydroxytryptamine1A (5-HT1A) receptor antagonist), but not SCH23390 (0.05 mg/kg, i.p.; a dopamine D1 receptor antagonist). Our findings suggested that the antidepressant-like effects of sanggenon G were mediated by an interaction with the serotonergic system. Further studies are needed to evaluate the potential of sanggenon G as an alternative therapeutic approach for the treatment of depression.

  8. Observations of parent-child co-shoppers in supermarkets: children's involvement in food selections, parental yielding, and refusal strategies.

    Science.gov (United States)

    O'Dougherty, Maureen; Story, Mary; Stang, Jamie

    2006-01-01

    The study aimed to collect descriptive information on the decision-making processes of adult shoppers around food purchases when young children are present. Anthropological field observations were conducted on adult-child grocery shoppers. Eleven supermarkets in the Minneapolis-St. Paul metropolitan region. A convenience sample (n = 142) of adult-child shoppers at 8 budget and 3 deluxe supermarkets located in diverse urban and suburban areas. Observations registered adult-child interactions over food selections, including parental yielding or refusal strategies and child engagement in shopping. Means and frequencies were calculated for food items considered. In 67 (50.4%) of the total 133 observations, a child initiated a request. Half (55.2%) of the requests were for sweets or snacks. Nearly half (47.8%) of adults yielded to the child's request. Brands and marketing techniques appeared to be a factor in 28.6% of selections. The most frequent adult refusals either provided an explanation or ignored the request. Adults yield to children's requests for sweets and snacks nearly as often as they refuse them. However, effective refusal strategies are used by many adults. Opportunities exist in the grocery store for adults to reinforce young children's interest in food and nutrition.

  9. Lignolytic Enzymes Production from Selected Mushrooms

    Directory of Open Access Journals (Sweden)

    H.M. Shantaveera Swamy

    2015-06-01

    Full Text Available In this paper, ligninase enzymes produced by selected mushrooms have been reported. We collected mushrooms from Western Ghats, most of them were edible food. Thirty samples isolated were tested using a plate assay through direct agar plate assay by using ABTS, decolourisation containing the fifteen isolates were able to decolourise the dye, indicating a lignin-degrading ability. Spectrophotometric enzyme assays from all selected isolates were carried out to examine the production of Ligninolytic enzymes (Laccase, lignin peroxidase and manganese peroxidase. Ten selected isolates produced all three kinds of enzymes tested. Lignolytic enzymes are groups of enzymes these are actively involved in bioremediation.

  10. 10 years of 1:1 taping in Belgium - A selection of murder cases involving fibre examination.

    Science.gov (United States)

    De Wael, K; Lepot, L; Lunstroot, K; Gason, F

    2016-01-01

    Since 2002, the 1:1 tape lifting technique on murder victims has been introduced in Belgium. Thirty-six murder cases for which a fibre examination was carried out in our laboratory between 2002 and 2012 are reviewed. These cases all involved the collection of microtraces on 39 victims using a 1:1 taping or similar technique, called semi 1:1 taping. The degree of "positive outcomes" for investigative and comparative fibre cases is high. Especially in cases where the modus operandi of suspects had to be verified, the examination of the 1:1 tape lifting led to very useful fibre results. The findings from some of these fibre cases are used to illustrate the advantages, drawbacks and points of interest of the two collection techniques. These are also compared with the zonal taping technique, another microtrace collection technique applied on victims at the crime scene.

  11. Homology modeling of mosquito cytochrome P450 enzymes involved in pyrethroid metabolism: insights into differences in substrate selectivity

    Directory of Open Access Journals (Sweden)

    Rongnoparut Pornpimol

    2011-09-01

    Full Text Available Abstract Background Cytochrome P450 enzymes (P450s have been implicated in insecticide resistance. Anopheles minumus mosquito P450 isoforms CYP6AA3 and CYP6P7 are capable of metabolizing pyrethroid insecticides, however CYP6P8 lacks activity against this class of compounds. Findings Homology models of the three An. minimus P450 enzymes were constructed using the multiple template alignment method. The predicted enzyme model structures were compared and used for molecular docking with insecticides and compared with results of in vitro enzymatic assays. The three model structures comprise common P450 folds but differences in geometry of their active-site cavities and substrate access channels are prominent. The CYP6AA3 model has a large active site allowing it to accommodate multiple conformations of pyrethroids. The predicted CYP6P7 active site is more constrained and less accessible to binding of pyrethroids. Moreover the predicted hydrophobic interface in the active-site cavities of CYP6AA3 and CYP6P7 may contribute to their substrate selectivity. The absence of CYP6P8 activity toward pyrethroids appears to be due to its small substrate access channel and the presence of R114 and R216 that may prevent access of pyrethroids to the enzyme heme center. Conclusions Differences in active site topologies among CYPAA3, CYP6P7, and CYP6P8 enzymes may impact substrate binding and selectivity. Information obtained using homology models has the potential to enhance the understanding of pyrethroid metabolism and detoxification mediated by P450 enzymes.

  12. Environmental selection pressures related to iron utilization are involved in the loss of the flavodoxin gene from the plant genome.

    Science.gov (United States)

    Pierella Karlusich, Juan J; Ceccoli, Romina D; Graña, Martín; Romero, Héctor; Carrillo, Néstor

    2015-02-16

    Oxidative stress and iron limitation represent the grim side of life in an oxygen-rich atmosphere. The versatile electron transfer shuttle ferredoxin, an iron-sulfur protein, is particularly sensitive to these hardships, and its downregulation under adverse conditions severely compromises survival of phototrophs. Replacement of ferredoxin by a stress-resistant isofunctional carrier, flavin-containing flavodoxin, is a widespread strategy employed by photosynthetic microorganisms to overcome environmental adversities. The flavodoxin gene was lost in the course of plant evolution, but its reintroduction in transgenic plants confers increased tolerance to environmental stress and iron starvation, raising the question as to why a genetic asset with obvious adaptive value was not kept by natural selection. Phylogenetic analyses reveal that the evolutionary history of flavodoxin is intricate, with several horizontal gene transfer events between distant organisms, including Eukarya, Bacteria, and Archaea. The flavodoxin gene is unevenly distributed in most algal lineages, with flavodoxin-containing species being overrepresented in iron-limited regions and scarce or absent in iron-rich environments. Evaluation of cyanobacterial genomic and metagenomic data yielded essentially the same results, indicating that there was little selection pressure to retain flavodoxin in iron-rich coastal/freshwater phototrophs. Our results show a highly dynamic evolution pattern of flavodoxin tightly connected to the bioavailability of iron. Evidence presented here also indicates that the high concentration of iron in coastal and freshwater habitats may have facilitated the loss of flavodoxin in the freshwater ancestor of modern plants during the transition of photosynthetic organisms from the open oceans to the firm land.

  13. Selection and evaluation of Debaryomyces hansenii isolates as potential bioprotective agents against toxigenic penicillia in dry-fermented sausages.

    Science.gov (United States)

    Núñez, Félix; Lara, María S; Peromingo, Belén; Delgado, Josué; Sánchez-Montero, Lourdes; Andrade, María J

    2015-04-01

    Biocontrol using autochthonous Debaryomyces hansenii isolates is a potentially suitable strategy for inhibiting toxigenic moulds in dry-cured meat products. The antifungal activity of 280 D. hansenii isolated from dry-cured meat products as well as the mode of action of the most active isolates against toxigenic penicillia were evaluated in this work. A 13.9% of the D. hansenii isolates showed inhibitory activity in a radial inhibition assay. The effects on penicillia growth of both the cell-free culture filtrate and volatile compounds from active yeast isolates were analysed. Penicillia growth inhibition by D. hansenii was probably based on additive or synergistic effects of several inhibiting factors such as competition for nutrient and space, and production of soluble or volatile compounds. When four D. hansenii isolates were tested on dry-fermented sausage, two of them produced a significantly growth reduction of the ochratoxigenic Penicillium verrucosum, keeping its counts under the level considered as hazardous for the mycotoxin presence. Therefore, the use of these two D. hansenii isolates during the processing of dry-fermented meat product could be a promising tool to control toxigenic moulds in the meat industry.

  14. "To-be-forgotten" statements become less true: Memory processes involved in selection and forgetting lead to truthfulness changes of ambiguous sentences.

    Science.gov (United States)

    Santos, Ana Sofia; Ramos, Tânia; Garcia-Marques, Leonel; Carneiro, Paula

    2017-06-01

    What happens when people try to forget something? What are the consequences of instructing people to intentionally forget a sentence? Recent studies employing the item-method directed forgetting paradigm have shown that to-be-forgotten (TBF) items are, in a subsequent task, emotionally devaluated relative to to-be-remembered (TBR) items, an aftereffect of memory selection (Vivas, Marful, Panagiotidou & Bajo, 2016). As such, distractor devaluation by attentional selection generalizes to memory selection. In this study, we use the item-method directed forgetting paradigm to test the effects of memory selection and inhibition on truth judgments of ambiguous sentences. We expected the relative standing of an item in the task (i.e., whether it was instructed to be remembered or forgotten) to affect the truthfulness value of that item, making TBF items less valid/truthful than TBR items. As predicted, ambiguous sentences associated with a "Forget" cue were subsequently judged as less true than sentences associated with a "Remember" cue, suggesting that instructions to intentionally forget a statement can produce changes in the validity/truthfulness of that statement. To our knowledge, this is the first study to show an influence of memory processes involved in selection and forgetting on the perceived truthfulness of sentences. © 2017 Scandinavian Psychological Associations and John Wiley & Sons Ltd.

  15. Selective isolation of potentially phosphate-mobilizing, biosurfactant-producing and biodegradative bacteria associated with a sub-Arctic, terricolous lichen, Peltigera membranacea.

    Science.gov (United States)

    Sigurbjörnsdóttir, Margrét Auður; Vilhelmsson, Oddur

    2016-06-01

    Lichens are the symbiotic association of fungi and a photosynthetic partner. However, non-phototrophic bacteria are also present and thought to comprise an essential part of the lichen symbiosis, although their roles in the symbiosis are still poorly understood. In this study, we isolated and characterized 110 non-phototrophic bacterial lichen associates from thalli of the terricolous lichen Peltigera membranacea The biodegradative and other nutrient-scavenging properties studied among selected isolates were phosphate mobilization, biosurfactant production and degradation of napthalene and several biopolymers, suggesting organic and inorganic nutrient scavenging as roles for bacteria in the lichen symbiotic association. Identification by partial 16S rRNA gene sequencing revealed that the isolates comprised 18 genera within the Proteobacteria, Actinobacteria, Bacteroidetes and Firmicutes, many with high similarities with bacteria typically associated with the plant and rhizosphere environments, could suggest that plants may be important sources of terricolous lichen-associated bacteria, or vice versa.

  16. A murine specific expansion of the Rhox cluster involved in embryonic stem cell biology is under natural selection

    Directory of Open Access Journals (Sweden)

    Keebler Jon

    2006-08-01

    Full Text Available Abstract Background The rodent specific reproductive homeobox (Rhox gene cluster on the X chromosome has been reported to contain twelve homeobox-containing genes, Rhox1-12. Results We have identified a 40 kb genomic region within the Rhox cluster that is duplicated eight times in tandem resulting in the presence of eight paralogues of Rhox2 and Rhox3 and seven paralogues of Rhox4. Transcripts have been identified for the majority of these paralogues and all but three are predicted to produce full-length proteins with functional potential. We predict that there are a total of thirty-two Rhox genes at this genomic location, making it the most gene-rich homoeobox cluster identified in any species. From the 95% sequence similarity between the eight duplicated genomic regions and the synonymous substitution rate of the Rhox2, 3 and 4 paralogues we predict that the duplications occurred after divergence of mouse and rat and represent the youngest homoeobox cluster identified to date. Molecular evolutionary analysis reveals that this cluster is an actively evolving region with Rhox2 and 4 paralogues under diversifying selection and Rhox3 evolving neutrally. The biological importance of this duplication is emphasised by the identification of an important role for Rhox2 and Rhox4 in regulating the initial stages of embryonic stem (ES cell differentiation. Conclusion The gene rich Rhox cluster provides the mouse with significant biological novelty that we predict could provide a substrate for speciation. Moreover, this unique cluster may explain species differences in ES cell derivation and maintenance between mouse, rat and human.

  17. Plant High-Affinity Potassium (HKT Transporters Involved in Salinity Tolerance: Structural Insights to Probe Differences in Ion Selectivity

    Directory of Open Access Journals (Sweden)

    Maria Hrmova

    2013-04-01

    Full Text Available High-affinity Potassium Transporters (HKTs belong to an important class of integral membrane proteins (IMPs that facilitate cation transport across the plasma membranes of plant cells. Some members of the HKT protein family have been shown to be critical for salinity tolerance in commercially important crop species, particularly in grains, through exclusion of Na+ ions from sensitive shoot tissues in plants. However, given the number of different HKT proteins expressed in plants, it is likely that different members of this protein family perform in a range of functions. Plant breeders and biotechnologists have attempted to manipulate HKT gene expression through genetic engineering and more conventional plant breeding methods to improve the salinity tolerance of commercially important crop plants. Successful manipulation of a biological trait is more likely to be effective after a thorough understanding of how the trait, genes and proteins are interconnected at the whole plant level. This article examines the current structural and functional knowledge relating to plant HKTs and how their structural features may explain their transport selectivity. We also highlight specific areas where new knowledge of plant HKT transporters is needed. Our goal is to present how knowledge of the structure of HKT proteins is helpful in understanding their function and how this understanding can be an invaluable experimental tool. As such, we assert that accurate structural information of plant IMPs will greatly inform functional studies and will lead to a deeper understanding of plant nutrition, signalling and stress tolerance, all of which represent factors that can be manipulated to improve agricultural productivity.

  18. Plant High-Affinity Potassium (HKT) Transporters involved in salinity tolerance: structural insights to probe differences in ion selectivity.

    Science.gov (United States)

    Waters, Shane; Gilliham, Matthew; Hrmova, Maria

    2013-04-09

    High-affinity Potassium Transporters (HKTs) belong to an important class of integral membrane proteins (IMPs) that facilitate cation transport across the plasma membranes of plant cells. Some members of the HKT protein family have been shown to be critical for salinity tolerance in commercially important crop species, particularly in grains, through exclusion of Na+ ions from sensitive shoot tissues in plants. However, given the number of different HKT proteins expressed in plants, it is likely that different members of this protein family perform in a range of functions. Plant breeders and biotechnologists have attempted to manipulate HKT gene expression through genetic engineering and more conventional plant breeding methods to improve the salinity tolerance of commercially important crop plants. Successful manipulation of a biological trait is more likely to be effective after a thorough understanding of how the trait, genes and proteins are interconnected at the whole plant level. This article examines the current structural and functional knowledge relating to plant HKTs and how their structural features may explain their transport selectivity. We also highlight specific areas where new knowledge of plant HKT transporters is needed. Our goal is to present how knowledge of the structure of HKT proteins is helpful in understanding their function and how this understanding can be an invaluable experimental tool. As such, we assert that accurate structural information of plant IMPs will greatly inform functional studies and will lead to a deeper understanding of plant nutrition, signalling and stress tolerance, all of which represent factors that can be manipulated to improve agricultural productivity.

  19. Isolation of novel single-chain Cro proteins targeted for binding to the bcl-2 transcription initiation site by repertoire selection and subunit combinatorics.

    Science.gov (United States)

    Jonas, Kristina; Van Der Vries, Erhard; Nilsson, Mikael T I; Widersten, Mikael

    2005-11-01

    New designed DNA-binding proteins may be recruited to act as transcriptional regulators and could provide new therapeutic agents in the treatment of genetic disorders such as cancer. We have isolated tailored DNA-binding proteins selected for affinity to a region spanning the transcription initiation site of the human bcl-2 gene. The proteins were derived from a single-chain derivative of the lambda Cro protein (scCro), randomly mutated in its recognition helices to construct libraries of protein variants of distinct DNA-binding properties. By phage display-afforded affinity selections combined with recombination of shuffled subunits, protein variants were isolated, which displayed high affinity for the target bcl-2 sequence, as determined by electrophoretic mobility shift and biosensor assays. The proteins analyzed were moderately sequence-specific but provide a starting point for further maturation of desired function.

  20. Ginsenoside Rd attenuates mitochondrial permeability transition and cytochrome C release in isolated spinal cord mitochondria: involvement of kinase-mediated pathways.

    Science.gov (United States)

    Zhou, Jin-Song; Wang, Jiang-Feng; He, Bao-Rong; Cui, Yong-Sheng; Fang, Xiang-Yi; Ni, Jian-Long; Chen, Jie; Wang, Kun-Zheng

    2014-06-03

    Ginsenoside Rd (Rd), one of the main active ingredients in Panax ginseng, has multifunctional activity via different mechanisms and neuroprotective effects that are exerted probably via its antioxidant or free radical scavenger action. However, the effects of Rd on spinal cord mitochondrial dysfunction and underlying mechanisms are still obscure. In this study, we sought to investigate the in vitro effects of Rd on mitochondrial integrity and redox balance in isolated spinal cord mitochondria. We verified that Ca2+ dissipated the membrane potential, provoked mitochondrial swelling and decreased NAD(P)H matrix content, which were all attenuated by Rd pretreatment in a dose-dependent manner. In contrast, Rd was not able to inhibit Ca2+ induced mitochondrial hydrogen peroxide generation. The results of Western blot showed that Rd significantly increased the expression of p-Akt and p-ERK, but had no effects on phosphorylation of PKC and p38. In addition, Rd treatment significantly attenuated Ca2+ induced cytochrome c release, which was partly reversed by antagonists of Akt and ERK, but not p-38 inhibitor. The effects of bisindolylmaleimide, a PKC inhibitor, on Rd-induced inhibition of cytochrome c release seem to be at the level of its own detrimental activity on mitochondrial function. Furthermore, we also found that pretreatment with Rd in vivo (10 and 50 mg/kg) protected spinal cord mitochondria against Ca2+ induced mitochondrial membrane potential dissipation and cytochrome c release. It is concluded that Rd regulate mitochondrial permeability transition pore formation and cytochrome c release through protein kinases dependent mechanism involving activation of intramitochondrial Akt and ERK pathways.

  1. Ginsenoside Rd Attenuates Mitochondrial Permeability Transition and Cytochrome c Release in Isolated Spinal Cord Mitochondria: Involvement of Kinase-Mediated Pathways

    Directory of Open Access Journals (Sweden)

    Jin-Song Zhou

    2014-06-01

    Full Text Available Ginsenoside Rd (Rd, one of the main active ingredients in Panax ginseng, has multifunctional activity via different mechanisms and neuroprotective effects that are exerted probably via its antioxidant or free radical scavenger action. However, the effects of Rd on spinal cord mitochondrial dysfunction and underlying mechanisms are still obscure. In this study, we sought to investigate the in vitro effects of Rd on mitochondrial integrity and redox balance in isolated spinal cord mitochondria. We verified that Ca2+ dissipated the membrane potential, provoked mitochondrial swelling and decreased NAD(PH matrix content, which were all attenuated by Rd pretreatment in a dose-dependent manner. In contrast, Rd was not able to inhibit Ca2+ induced mitochondrial hydrogen peroxide generation. The results of Western blot showed that Rd significantly increased the expression of p-Akt and p-ERK, but had no effects on phosphorylation of PKC and p38. In addition, Rd treatment significantly attenuated Ca2+ induced cytochrome c release, which was partly reversed by antagonists of Akt and ERK, but not p-38 inhibitor. The effects of bisindolylmaleimide, a PKC inhibitor, on Rd-induced inhibition of cytochrome c release seem to be at the level of its own detrimental activity on mitochondrial function. Furthermore, we also found that pretreatment with Rd in vivo (10 and 50 mg/kg protected spinal cord mitochondria against Ca2+ induced mitochondrial membrane potential dissipation and cytochrome c release. It is concluded that Rd regulate mitochondrial permeability transition pore formation and cytochrome c release through protein kinases dependent mechanism involving activation of intramitochondrial Akt and ERK pathways.

  2. Detection of Extended Spectrum Beta-Lactamases Resistance Genes among Bacteria Isolated from Selected Drinking Water Distribution Channels in Southwestern Nigeria

    Science.gov (United States)

    Ogunjobi, Adeniyi A.

    2016-01-01

    Extended Spectrum Beta-Lactamases (ESBL) provide high level resistance to beta-lactam antibiotics among bacteria. In this study, previously described multidrug resistant bacteria from raw, treated, and municipal taps of DWDS from selected dams in southwestern Nigeria were assessed for the presence of ESBL resistance genes which include blaTEM, blaSHV, and blaCTX by PCR amplification. A total of 164 bacteria spread across treated (33), raw (66), and municipal taps (68), belonging to α-Proteobacteria, β-Proteobacteria, γ-Proteobacteria, Flavobacteriia, Bacilli, and Actinobacteria group, were selected for this study. Among these bacteria, the most commonly observed resistance was for ampicillin and amoxicillin/clavulanic acid (61 isolates). Sixty-one isolates carried at least one of the targeted ESBL genes with blaTEM being the most abundant (50/61) and blaCTX being detected least (3/61). Klebsiella was the most frequently identified genus (18.03%) to harbour ESBL gene followed by Proteus (14.75%). Moreover, combinations of two ESBL genes, blaSHV + blaTEM or blaCTX + blaTEM, were observed in 11 and 1 isolate, respectively. In conclusion, classic blaTEM ESBL gene was present in multiple bacterial strains that were isolated from DWDS sources in Nigeria. These environments may serve as foci exchange of genetic traits in a diversity of Gram-negative bacteria. PMID:27563674

  3. Isolation of novel non-HLA gene fragments from the hemochromatosis region (6p21. 3) by cDNA hybridization selection

    Energy Technology Data Exchange (ETDEWEB)

    Goei, V.L.; Capossela, A.; Gruen, J.R.; Parimoo, S.; Chu, T.W. (Yale Univ. School of Medicine, New Haven, CT (United States))

    1994-02-01

    It has previously been shown that cDNA hybridization selection can identify and recover novel genes from large cloned genomic DNA such as cosmids or YACs. In an effort to identify candidate genes for hemochromatosis, this technique was applied to a 320-kb YAC containing the HLA-A gene. A short fragment cDNA library derived from human duodenum was selected with the YAC DNA. Ten novel gene fragments were isolated, characterized, and localized on the physical map of the YAC. 39 refs., 4 figs., 3 tabs.

  4. Contrasting signals of positive selection in genes involved in human skin-color variation from tests based on SNP scans and resequencing

    Directory of Open Access Journals (Sweden)

    de Gruijter Johanna

    2011-12-01

    Full Text Available Abstract Background Numerous genome-wide scans conducted by genotyping previously ascertained single-nucleotide polymorphisms (SNPs have provided candidate signatures for positive selection in various regions of the human genome, including in genes involved in pigmentation traits. However, it is unclear how well the signatures discovered by such haplotype-based test statistics can be reproduced in tests based on full resequencing data. Four genes (oculocutaneous albinism II (OCA2, tyrosinase-related protein 1 (TYRP1, dopachrome tautomerase (DCT, and KIT ligand (KITLG implicated in human skin-color variation, have shown evidence for positive selection in Europeans and East Asians in previous SNP-scan data. In the current study, we resequenced 4.7 to 6.7 kb of DNA from each of these genes in Africans, Europeans, East Asians, and South Asians. Results Applying all commonly used neutrality-test statistics for allele frequency distribution to the newly generated sequence data provided conflicting results regarding evidence for positive selection. Previous haplotype-based findings could not be clearly confirmed. Although some tests were marginally significant for some populations and genes, none of them were significant after multiple-testing correction. Combined P values for each gene-population pair did not improve these results. Application of Approximate Bayesian Computation Markov chain Monte Carlo based to these sequence data using a simple forward simulator revealed broad posterior distributions of the selective parameters for all four genes, providing no support for positive selection. However, when we applied this approach to published sequence data on SLC45A2, another human pigmentation candidate gene, we could readily confirm evidence for positive selection, as previously detected with sequence-based and some haplotype-based tests. Conclusions Overall, our data indicate that even genes that are strong biological candidates for positive

  5. Evaluation of Two Standard and Two Chromogenic Selective Media for Optimal Growth and Enumeration of Isolates of 16 Unique Bacillus Species.

    Science.gov (United States)

    Kabir, M Shahjahan; Hsieh, Ying-Hsin; Simpson, Steven; Kerdahi, Khalil; Sulaiman, Irshad M

    2017-06-01

    The genus Bacillus is a group of gram-positive endospore-forming bacteria that can cause food poisoning and diarrheal illness in humans. A wide range of food products have been linked to foodborne outbreaks associated with these opportunistic pathogens. The U.S. Food and Drug Administration recommends (in their Bacteriological Analytical Manual) the use of Bacara or mannitol egg yolk polymyxin (MYP) agar plates and the most-probable-number (MPN) method for enumeration and confirmation of Bacillus cereus and related species isolated from foods, sporadic cases, outbreaks, and routine environmental surveillance samples. We performed a comparative analysis of two chromogenic media (Bacara and Brilliance) and two traditional media (MYP and polymyxin egg yolk mannitol bromothymol blue agar [PEMBA]) for the isolation and enumeration of 16 Bacillus species under modified growth conditions that included pH, temperature, and dilution factor. A total of 50 environmental, food, and American Type Culture Collection reference isolates from 16 distinct Bacillus species were evaluated. A food adulteration experiment also was carried out by artificially adulterating two baby food matrices with two isolates each of B. cereus and Bacillus thuringiensis . Our results clearly indicated that chromogenic plating media (Bacara and Brilliance) are better than conventional standard media (MYP and PEMBA) for the detection and enumeration of B. cereus in foods and other official regulatory samples. The comparison of the two chromogenic media also indicated that Brilliance medium to be more efficient and selective for the isolation of Bacillus.

  6. Screening of Lactobacillus isolates from gastrointestinal tract of guinea fowl for probiotic qualities using in vitro tests to select species-specific probiotic candidates.

    Science.gov (United States)

    Vineetha, P G; Tomar, S; Saxena, V K; Susan, C; Sandeep, S; Adil, K; Mukesh, K

    2016-08-01

    A total of 32 Lactobacillus isolates, 8 each from the crop (LGFCP1-LGFCP8), proventriculus (LGFP9-LGFP16), ileum (LGFI17-LGFI24) and caeca (LGFCM25-LGFCM32) were isolated from 25 adult guinea fowl (Pearl variety), 22-28 weeks of age, and characterised morphologically, physiologically, biochemically and by molecular methods. Isolates were screened for their probiotic quality using range of in vitro tests: aggregation test, cell surface hydrophobicity, resistance to bile salts and acidic conditions, enzymatic tests and coaggregation and antagonistic test. Based on in vitro test results and a novel scoring method, the two best isolates were selected and partial 16S rRNA sequencing was done. BLAST (Basic Local Alignment Search Tool) analysis of sequence of isolate LGFCP4 showed 99% genetic identity with Lactobacillus plantarum and LGFP16 with Lactobacillus reuteri. The study shows that these two microbial agents may be suitable as potential probiotic candidates in guinea fowl, as well as in a feed supplement for other poultry species.

  7. Genotypic Characterization of Azotobacteria Isolated from Argentinean Soils and Plant-Growth-Promoting Traits of Selected Strains with Prospects for Biofertilizer Production

    Directory of Open Access Journals (Sweden)

    Esteban Julián Rubio

    2013-01-01

    Full Text Available The genetic diversity among 31 putative Azotobacter isolates obtained from agricultural and non-agricultural soils was assessed using rep-PCR genomic fingerprinting and identified to species level by ARDRA and partial 16S rRNA gene sequence analysis. High diversity was found among the isolates, identified as A. chroococcum, A. salinestris, and A. armeniacus. Selected isolates were characterized on the basis of phytohormone biosynthesis, nitrogenase activity, siderophore production, and phosphate solubilization. Indole-3 acetic-acid (IAA, gibberellin (GA3 and zeatin (Z biosynthesis, nitrogenase activity, and siderophore production were found in all evaluated strains, with variation among them, but no phosphate solubilization was detected. Phytohormones excreted to the culture medium ranged in the following concentrations: 2.2–18.2 μg IAA mL−1, 0.3–0.7 μg GA3 mL−1, and 0.5–1.2 μg Z mL−1. Seed inoculations with further selected Azotobacter strains and treatments with their cell-free cultures increased the number of seminal roots and root hairs in wheat seedlings. This latter effect was mimicked by treatments with IAA-pure solutions, but it was not related to bacterial root colonization. Our survey constitutes a first approach to the knowledge of Azotobacter species inhabiting Argentinean soils in three contrasting geographical regions. Moreover, this phenotypic characterization constitutes an important contribution to the selection of Azotobacter strains for biofertilizer formulations.

  8. Restoring the selectivity of modified charcoal cefoperazone deoxycholate agar for the isolation of Campylobacter species using tazobactam, a β-lactamase inhibitor.

    Science.gov (United States)

    Smith, Shaun; Meade, Joseph; McGill, Kevina; Gibbons, James; Bolton, Declan; Whyte, Paul

    2015-10-01

    Extended spectrum β-lactamase (ESBL) producing Escherichia coli have emerged as a contaminant on modified charcoal cefoperazone deoxycholate agar (mCCDA) when attempting to selectively isolate Campylobacter spp. from poultry. E. coli are particularly problematic given their ability to grow under microaerophilic conditions and have been shown to outcompete Campylobacter species making Campylobacter detection or enumeration difficult. This paper recommends a novel method for restoring the selectivity of mCCDA using tazobactam, a β-lactamase inhibitor. The method significantly inhibited ESBL E. coli growth in spiked or naturally contaminated broiler caecal samples (p≤0.01) when compared to conventional mCCDA. This effect was seen at concentrations as low as 1mg/L tazobactam. TmCCDA(1) was found to inhibit up to 8 log10 CFU/mL of ESBL E. coli in mixed pure cultures and 7.5 log10 CFU/mL in caecal samples. Furthermore TmCCDA concentrations up to 10 mg/L had no statistically significant inhibitory effect (p≥0.05) on the recovery of a panel of 27 Campylobacter jejuni and 5 Campylobacter coli isolates when compared to conventional mCCDA. From this study it is suggested that tazobactam, which is more chemically stable than clavulanic acid or sulbactam, is more suitable for restoring the selectivity of mCCDA for the detection or isolation of campylobacters.

  9. Genotypic characterization of Azotobacteria isolated from Argentinean soils and plant-growth-promoting traits of selected strains with prospects for biofertilizer production.

    Science.gov (United States)

    Rubio, Esteban Julián; Montecchia, Marcela Susana; Tosi, Micaela; Cassán, Fabricio Darío; Perticari, Alejandro; Correa, Olga Susana

    2013-01-01

    The genetic diversity among 31 putative Azotobacter isolates obtained from agricultural and non-agricultural soils was assessed using rep-PCR genomic fingerprinting and identified to species level by ARDRA and partial 16S rRNA gene sequence analysis. High diversity was found among the isolates, identified as A. chroococcum, A. salinestris, and A. armeniacus. Selected isolates were characterized on the basis of phytohormone biosynthesis, nitrogenase activity, siderophore production, and phosphate solubilization. Indole-3 acetic-acid (IAA), gibberellin (GA3) and zeatin (Z) biosynthesis, nitrogenase activity, and siderophore production were found in all evaluated strains, with variation among them, but no phosphate solubilization was detected. Phytohormones excreted to the culture medium ranged in the following concentrations: 2.2-18.2 μ g IAA mL(-1), 0.3-0.7 μ g GA3 mL(-1), and 0.5-1.2 μ g Z mL(-1). Seed inoculations with further selected Azotobacter strains and treatments with their cell-free cultures increased the number of seminal roots and root hairs in wheat seedlings. This latter effect was mimicked by treatments with IAA-pure solutions, but it was not related to bacterial root colonization. Our survey constitutes a first approach to the knowledge of Azotobacter species inhabiting Argentinean soils in three contrasting geographical regions. Moreover, this phenotypic characterization constitutes an important contribution to the selection of Azotobacter strains for biofertilizer formulations.

  10. Isolation of Alcaligenes sp strain L6 at low oxygen concentrations and degradation of 3-chlorobenzoate via a pathway not involving (chloro)catechols

    NARCIS (Netherlands)

    Krooneman, J; Wieringa, EBA; Moore, ERB; Gerritse, J; Prins, RA; Gottschal, JC

    1996-01-01

    Isolations of 3-chlorobenzoate (3CBA)-degrading aerobic bacteria under reduced O-2, partial pressures yielded organisms which metabolized 3CBA via the gentisate or the protocatechuate pathway rather than via the catechol route. The 3CBA metabolism of one of these isolates, L6, which,vas identified a

  11. Identification of amino acid changes in the envelope glycoproteins of bovine viral diarrhea viruses isolated from alpaca that may be involved in host adaptation

    Science.gov (United States)

    Bovine viral diarrhea viruses (BVDV) are most commonly associated with infections of cattle. However, BVDV is often isolated from closely related ruminants with a number of BVDV-1b viruses being isolated from alpacas that were both acutely and persistently infected (PI). The complete nucleotide se...

  12. Natural selection drives the fine-scale divergence of a coevolutionary arms race involving a long-mouthed weevil and its obligate host plant

    Directory of Open Access Journals (Sweden)

    Toju Hirokazu

    2009-01-01

    Full Text Available Abstract Background One of the major recent advances in evolutionary biology is the recognition that evolutionary interactions between species are substantially differentiated among geographic populations. To date, several authors have revealed natural selection pressures mediating the geographically-divergent processes of coevolution. How local, then, is the geographic structuring of natural selection in coevolutionary systems? Results I examined the spatial scale of a "geographic selection mosaic," focusing on a system involving a seed-predatory insect, the camellia weevil (Curculio camelliae, and its host plant, the Japanese camellia (Camellia japonica. In this system, female weevils excavate camellia fruits with their extremely-long mouthparts to lay eggs into seeds, while camellia seeds are protected by thick pericarps. Quantitative evaluation of natural selection demonstrated that thicker camellia pericarps are significantly favored in some, but not all, populations within a small island (Yakushima Island, Japan; diameter ca. 30 km. At the extreme, camellia populations separated by only several kilometers were subject to different selection pressures. Interestingly, in a population with the thickest pericarps, camellia individuals with intermediate pericarp thickness had relatively high fitness when the potential costs of producing thick pericarps were considered. Also importantly, some parameters of the weevil - camellia interaction such as the severity of seed infestation showed clines along temperature, suggesting the effects of climate on the fine-scale geographic differentiation of the coevolutionary processes. Conclusion These results show that natural selection can drive the geographic differentiation of interspecific interactions at surprisingly small spatial scales. Future studies should reveal the evolutionary/ecological outcomes of the "fine scale geographic mosaics" in biological communities.

  13. Pattern of Bacterial Pathogens and Their Susceptibility Isolated from Surgical Site Infections at Selected Referral Hospitals, Addis Ababa, Ethiopia

    Directory of Open Access Journals (Sweden)

    Walelign Dessie

    2016-01-01

    Full Text Available Background. The emergence of multidrug resistant bacterial pathogens in hospitals is becoming a challenge for surgeons to treat hospital acquired infections. Objective. To determine bacterial pathogens and drug susceptibility isolated from surgical site infections at St. Paul Specialized Hospital Millennium Medical College and Yekatit 12 Referral Hospital Medical College, Addis Ababa, Ethiopia. Methods. A cross-sectional study was conducted between October 2013 and March 2014 on 107 surgical site infected patients. Wound specimens were collected using sterile cotton swab and processed as per standard operative procedures in appropriate culture media; and susceptibility testing was done using Kirby-Bauer disc diffusion technique. The data were analyzed by using SPSS version 20. Result. From a total of 107 swabs collected, 90 (84.1% were culture positive and 104 organisms were isolated. E. coli (24 (23.1% was the most common organism isolated followed by multidrug resistant Acinetobacter species (23 (22.1%. More than 58 (75% of the Gram negative isolates showed multiple antibiotic resistance (resistance ≥ 5 drugs. Pan-antibiotic resistance was noted among 8 (34.8% Acinetobacter species and 3 (12.5% E. coli. This calls for abstinence from antibiotic abuse. Conclusion. Gram negative bacteria were the most important isolates accounting for 76 (73.1%. Ampicillin, amoxicillin, penicillin, cephazoline, and tetracycline showed resistance while gentamicin and ciprofloxacin were relatively effective antimicrobials.

  14. Characterization of native Bacillus thuringiensis strains and selection of an isolate active against Spodoptera frugiperda and Peridroma saucia.

    Science.gov (United States)

    Alvarez, Analía; Virla, Eduardo G; Pera, Licia M; Baigorí, Mario D

    2009-12-01

    Twelve Bacillus thuringiensis (Bt) strains, isolated from larvae and soil samples in Argentina, were molecularly and phenotypically characterized and their insecticidal activities against Spodoptera frugiperda and Peridroma saucia were determined. One isolate--Bt RT--produced more than 93% mortality on first instar larvae of both species, which was higher than that produced by the reference strain Bt 4D1. Bt RT carried a different cry gene profile than Bt 4D1. Scanning electron microscopy showed the presence of bipyramidal and cuboidal crystals. Phenotypic characterization revealed lytic enzymes that could contribute to Bt pathogenicity.

  15. Selective block of IKs plays a significant role in MAP triangulation induced by IKr block in isolated rabbit heart.

    Science.gov (United States)

    Guérard, N C; Traebert, M; Suter, W; Dumotier, B M

    2008-01-01

    The role of IKr (rapidly-activating delayed rectifier K(+) current) block in triangulation of monophasic action potentials (MAP) and in development of torsade de pointes (TdP) arrhythmia is known. Combined IKr and IKs (slowly-activating delayed rectifier K(+) current) block has been demonstrated to promote TdP. The aim of this study was to describe a possible implication of IKs block in MAP triangulation. Four contact electrodes were placed on the epicardium of the left ventricle of Langendorff-perfused rabbit hearts to record monophasic action potentials (MAP), with an IKr blocker d,l-sotalol (3 to 100 microM, n=6) or a non-selective IKr blocker, quinidine (1 to 30 microM, n=6). Their effects were assessed with or without a specific IKs blocker chromanol 293B (20 microM, n=6), on MAP duration at 30, 60 and 90% of repolarization (APD30, 60 and 90, respectively) and MAP triangulation (APD90-APD30) at 1 and 0.2 Hz. D,L-sotalol increased significantly APD90 and triangulation with reverse use-dependency for concentrations > or =10 microM. Quinidine markedly prolonged APD90 and triangulation with reverse use-dependency at concentrations > or =3 microM. Chromanol 293B alone had no effects on APD, but when combined with D,L-sotalol or quinidine (i) increased APD prolonging effects, (ii) lowered values of pro-arrhythmic concentrations, (iii) increased incidence and length of D,L-sotalol- or quinidine-induced Early Afterdepolarizations (EADs) and TdP. All these events were primarily due to an important slowing of final repolarization, i.e. a marked increased triangulation. IKs, even of low amplitude in rabbits, plays a key role in ventricular repolarization. IKs is involved in prolonged MAP duration mainly by triangulation and subsequent increased drug arrhythmogenicity. Therefore drug affinity for IKs must be evaluated with IKr studies as part of preclinical drug cardiac safety assessment.

  16. Antidepressant-like effect of ursolic acid isolated from Rosmarinus officinalis L. in mice: evidence for the involvement of the dopaminergic system.

    Science.gov (United States)

    Machado, D G; Neis, V B; Balen, G O; Colla, A; Cunha, M P; Dalmarco, J B; Pizzolatti, M G; Prediger, R D; Rodrigues, A L S

    2012-12-01

    Ursolic acid, a constituent from Rosmarinus officinalis, is a triterpenoid compound which has been extensively known for its anticancer and antioxidant properties. In the present study, we investigated the antidepressant-like effect of ursolic acid isolated from this plant in two predictive tests of antidepressant property, the tail suspension test (TST) and the forced swimming test (FST) in mice. Furthermore, the involvement of dopaminergic system in its antidepressant-like effect was investigated in the TST. Ursolic acid reduced the immobility time in the TST (0.01 and 0.1mg/kg, p.o.) and in the FST (10mg/kg, p.o.), similar to fluoxetine (10mg/kg, p.o.), imipramine (1mg/kg, p.o.) and bupropion (10mg/kg, p.o.). The effect of ursolic acid (0.1mg/kg, p.o.) in the TST was prevented by the pretreatment of mice with SCH23390 (0.05mg/kg, s.c., a dopamine D(1) receptor antagonist) and sulpiride (50mg/kg, i.p., a dopamine D(2) receptor antagonist). The administration of a sub-effective dose of ursolic acid (0.001mg/kg, p.o.) in combination with sub-effective doses of SKF38393 (0.1mg/kg, s.c., a dopamine D(1) receptor agonist), apomorphine (0.5μg/kg, i.p., a preferential dopamine D(2) receptor agonist) or bupropion (1mg/kg, i.p., a dual dopamine/noradrenaline reuptake inhibitor) reduced the immobility time in the TST as compared with either drug alone. Ursolic acid and dopaminergic agents alone or in combination did not cause significant alterations in the locomotor and exploratory activities. These results indicate that the antidepressant-like effect of ursolic acid in the TST is likely mediated by an interaction with the dopaminergic system, through the activation of dopamine D(1) and D(2) receptors.

  17. Analysis of the complete sequences of two biologically distinct Zucchini yellow mosaic virus isolates further evidences the involvement of a single amino acid in the virus pathogenicity.

    Science.gov (United States)

    Nováková, S; Svoboda, J; Glasa, M

    2014-01-01

    The complete genome sequences of two Slovak Zucchini yellow mosaic virus isolates (ZYMV-H and ZYMV-SE04T) were determined. These isolates differ significantly in their pathogenicity, producing either severe or very mild symptoms on susceptible cucurbit hosts. The viral genome of both isolates consisted of 9593 nucleotides in size, and contained an open reading frame encoding a single polyprotein of 3080 amino acids. Despite their different biological properties, an extremely high nucleotide identity could be noted (99.8%), resulting in differences of only 5 aa, located in the HC-Pro, P3, and NIb, respectively. In silico analysis including 5 additional fully-sequenced and phylogenetically closely-related isolates known to induce different symptoms in cucurbits was performed. This suggested that the key single mutation responsible for virus pathogenicity is likely located in the N-terminal part of P3, adjacent to the PIPO.

  18. Spatially heterogeneous environmental selection strengthens evolution of reproductively isolated populations in a Dobzhansky-Muller system of hybrid incompatibility

    Science.gov (United States)

    Samuel A. Cushman; Erin L. Landguth

    2016-01-01

    Within-species hybrid incompatibility can arise when combinations of alleles at more than one locus have low fitness but where possession of one of those alleles has little or no fitness consequence for the carriers. Limited dispersal with small numbers of mate potentials alone can lead to the evolution of clusters of reproductively isolated genotypes despite...

  19. Selection for protection from insolation results in the visual isolation of Yellow-eyed Penguin Megadyptes antipodes nests

    CSIR Research Space (South Africa)

    Clark, RD

    2015-06-01

    Full Text Available The concealed and widely dispersed nests of the rare and endangered Yellow-eyed Penguin Megadyptes antipodes, or “hoiho”, have been considered to reflect an essential requirement for the visual isolation of nest sites from conspecifics. However...

  20. Infectivity analysis of a blackgram isolate of Mungbean yellow mosaic virus and genetic assortment with MYMIV in selective hosts.

    Science.gov (United States)

    Haq, Q M I; Rouhibakhsh, A; Ali, Arif; Malathi, V G

    2011-06-01

    Yellow mosaic disease in grain legumes in Indian subcontinent is caused by two important virus species viz. Mungbean yellow mosaic virus (MYMV) and Mungbean yellow mosaic India virus (MYMIV), belonging to the genus Begomovirus of the family Geminiviridae. The genomic components of a begomovirus causing yellow mosaic disease in blackgram in southern India were cloned and sequenced. Nucleotide sequence comparison of DNA A component shows the virus isolate to be a variant of Mungbean yellow mosaic virus:-(MYMV-[IN:Vam:05]). However, DNA B component of the present virus isolate has greater similarity (92%) to Mungbean yellow mosaic India virus. Agroinoculations of the viral clones produced typical yellow mosaic symptoms in blackgram and mungbean, severe leaf curl and stunting in French bean, similar to blackgram isolate of MYMIV. Blackgram isolates of both the virus species were only mildly infectious on cowpea, produced atypical leaf curl symptoms and not yellow or golden mosaic. In agroinoculations done by exchanging genomic components, symptom expression was seen only in French bean. In cowpea, blackgram and mungbean there was no visible symptoms though viral DNA could be detected by PCR.

  1. VapB type 8 plasmids in Rhodococcus equi isolated from the small intestine of pigs and comparison of selective culture media.

    Science.gov (United States)

    Lara, G H B; Takai, S; Sasaki, Y; Kakuda, T; Listoni, F J P; Risseti, R M; de Morais, A B C; Ribeiro, M G

    2015-09-01

    The virulence-plasmid profile of Rhodococcus equi strains isolated from Suidae and humans is similar. Recent evidence suggests that the consumption of pork products contaminated with faeces might be a potential source of R. equi infections in humans, mainly to patients with rhodococcosis without history of contact with pigs or pig farms. This study investigated the virulence-associated genes (vapA and vapB) and plasmid profiles of R. equi among the 150 samples of small intestinal content obtained from slaughtered pigs. In addition, all samples were subjected to microbiological culture in conventional sheep blood agar and CAZ-NB, TCP and TVP selective media. A total of 40 (26·7%) of the samples recovered R. equi, with two samples recovering isolates harbouring the VapB type 8 plasmid. Among the 150 pigs sampled herein, CAZ-NB was considered the best selective medium for the isolation of R. equi from faeces. Our results provide evidence that the contamination of slaughtered pig carcasses with pathogenic R. equi might occur through faeces, representing a public health concern. Furthermore, this study is the first description of R. equi strains carrying the VapB plasmid in the gut of pigs. Intermediately virulent (VapB) is a common plasmid-type harboured by R. equi isolated from pigs and humans with AIDS. Curiously, humans with rhodococcosis usually have no history of contact with pigs or pig farms. Virulence-plasmid profile of 40 R. equi isolated among 150 small intestine content samples from pigs revelled two carrying isolates with the VapB type-8 plasmids. Moreover, comparison of three selective culture media shows that CAZ-NB was the best. Our results provide evidence that contamination of slaughtered pig carcasses with pathogenic R. equi might occur through faeces, representing a public health concern. Furthermore, R. equi carrying VapB type-8 plasmids types are described for the first time in the gut of the pig. © 2015 The Society for Applied

  2. Geometric morphometrics of nine field isolates of Aedes aegypti with different resistance levels to lambda-cyhalothrin and relative fitness of one artificially selected for resistance.

    Science.gov (United States)

    Jaramillo-O, Nicolás; Fonseca-González, Idalyd; Chaverra-Rodríguez, Duverney

    2014-01-01

    Aedes aegypti, a mosquito closely associated with humans, is the principal vector of dengue virus which currently infects about 400 million people worldwide. Because there is no way to prevent infection, public health policies focus on vector control; but insecticide-resistance threatens them. However, most insecticide-resistant mosquito populations exhibit fitness costs in absence of insecticides, although these costs vary. Research on components of fitness that vary with insecticide-resistance can help to develop policies for effective integrated management and control. We investigated the relationships in wing size, wing shape, and natural resistance levels to lambda-cyhalothrin of nine field isolates. Also we chose one of these isolates to select in lab for resistance to the insecticide. The main life-traits parameters were assessed to investigate the possible fitness cost and its association with wing size and shape. We found that wing shape, more than wing size, was strongly correlated with resistance levels to lambda-cyhalothrin in field isolates, but founder effects of culture in the laboratory seem to change wing shape (and also wing size) more easily than artificial selection for resistance to that insecticide. Moreover, significant fitness costs were observed in response to insecticide-resistance as proved by the diminished fecundity and survival of females in the selected line and the reversion to susceptibility in 20 generations of the non-selected line. As a practical consequence, we think, mosquito control programs could benefit from this knowledge in implementing efficient strategies to prevent the evolution of resistance. In particular, the knowledge of reversion to susceptibility is important because it can help in planning better strategies of insecticide use to keep useful the few insecticide-molecules currently available.

  3. A novel 9 × 9 map-based solvent selection strategy for targeted counter-current chromatography isolation of natural products.

    Science.gov (United States)

    Liang, Junling; Meng, Jie; Wu, Dingfang; Guo, Mengzhe; Wu, Shihua

    2015-06-26

    Counter-current chromatography (CCC) is an efficient liquid-liquid chromatography technique for separation and purification of complex mixtures like natural products extracts and synthetic chemicals. However, CCC is still a challenging process requiring some special technical knowledge especially in the selection of appropriated solvent systems. In this work, we introduced a new 9 × 9 map-based solvent selection strategy for CCC isolation of targets, which permit more than 60 hexane-ethyl acetate-methanol-water (HEMWat) solvent systems as the start candidates for the selection of solvent systems. Among these solvent systems, there are clear linear correlations between partition coefficient (K) and the system numbers. Thus, an appropriate CCC solvent system (i.e., sweet spot for K = 1) may be hit by measurement of k values of the target only in two random solvent systems. Besides this, surprisingly, we found that through two sweet spots, we could get a line ("Sweet line") where there are infinite sweet solvent systems being suitable for CCC separation. In these sweet solvent systems, the target has the same partition coefficient (K) but different solubilities. Thus, the better sweet solvent system with higher sample solubility can be obtained for high capacity CCC preparation. Furthermore, we found that there is a zone ("Sweet zone") where all solvent systems have their own sweet partition coefficients values for the target in range of 0.4 natural products as standards and further confirmed by isolation of several targets including honokiol and magnolol from the extracts of Magnolia officinalis Rehd. Et Wils and tanshinone IIA from Salvia miltiorrhiza Bunge. In practice, it is much easier to get a suitable solvent system only by making a simple screening two to four HEMWat two-phase solvent systems to obtain the sweet line or sweet zone without special knowledge or comprehensive standards as references. This is an important advancement for solvent system selection

  4. Genetic polymorphism and effect of natural selection at domain I of apical membrane antigen-1 (AMA-1) in Plasmodium vivax isolates from Myanmar.

    Science.gov (United States)

    Moon, Sung-Ung; Na, Byoung-Kuk; Kang, Jung-Mi; Kim, Jung-Yeon; Cho, Shin-Hyeong; Park, Yun-Kyu; Sohn, Woon-Mok; Lin, Khin; Kim, Tong-Soo

    2010-05-01

    Malaria is endemic or hypoendemic in Myanmar and the country still contributes to the high level of malaria deaths in South-East Asia. Although information on the nature and extent of population diversity within malaria parasites in the country is essential not only for understanding the epidemic situation but also to establish a proper control strategy, very little data is currently available on the extent of genetic polymorphisms of the malaria parasites in Myanmar. In this study, we analyzed the genetic polymorphism and natural selection at domain I of the apical membrane antigen-1 (AMA-1) among Plasmodium vivax Myanmar isolates. A total of 34 distinguishable haplotypes were identified among the 76 isolates sequenced. Comparison with the previously available PvAMA-1 sequences in the GenBank database revealed that 21 of them were new haplotypes that have never been reported till date. The difference between the rate of nonsynonymous (dN) and synonymous (dS) mutations was positive (dN-dS, 0.013+/-0.005), suggesting the domain I is under positive natural selection. The Tajima's D statistics was found to be -0.74652, suggesting that the gene has evolved under population size expansion and/or positive selection. The minimum recombination events were also high, indicating that recombination may occur within the domain I resulting in allelic diversity of PvAMA-1. Our results collectively suggest that PvAMA-1 displays high genetic polymorphism among Myanmar P. vivax isolates with highly diversifying selection at domain I. These results have significant implications in understanding the nature of P. vivax population circulating in Myanmar as well as providing useful information for malaria vaccine development based on this antigen.

  5. Geometric morphometrics of nine field isolates of Aedes aegypti with different resistance levels to lambda-cyhalothrin and relative fitness of one artificially selected for resistance.

    Directory of Open Access Journals (Sweden)

    Nicolás Jaramillo-O

    Full Text Available Aedes aegypti, a mosquito closely associated with humans, is the principal vector of dengue virus which currently infects about 400 million people worldwide. Because there is no way to prevent infection, public health policies focus on vector control; but insecticide-resistance threatens them. However, most insecticide-resistant mosquito populations exhibit fitness costs in absence of insecticides, although these costs vary. Research on components of fitness that vary with insecticide-resistance can help to develop policies for effective integrated management and control. We investigated the relationships in wing size, wing shape, and natural resistance levels to lambda-cyhalothrin of nine field isolates. Also we chose one of these isolates to select in lab for resistance to the insecticide. The main life-traits parameters were assessed to investigate the possible fitness cost and its association with wing size and shape. We found that wing shape, more than wing size, was strongly correlated with resistance levels to lambda-cyhalothrin in field isolates, but founder effects of culture in the laboratory seem to change wing shape (and also wing size more easily than artificial selection for resistance to that insecticide. Moreover, significant fitness costs were observed in response to insecticide-resistance as proved by the diminished fecundity and survival of females in the selected line and the reversion to susceptibility in 20 generations of the non-selected line. As a practical consequence, we think, mosquito control programs could benefit from this knowledge in implementing efficient strategies to prevent the evolution of resistance. In particular, the knowledge of reversion to susceptibility is important because it can help in planning better strategies of insecticide use to keep useful the few insecticide-molecules currently available.

  6. Determination of the Influence of Substrate Concentration on Enzyme Selectivity Using Whey Protein Isolate and Bacillus licheniformis Protease

    NARCIS (Netherlands)

    Butré, C.I.; Sforza, S.; Gruppen, H.; Wierenga, P.A.

    2014-01-01

    Increasing substrate concentration during enzymatic protein hydrolysis results in a decrease in hydrolysis rate. To test if changes in the mechanism of hydrolysis also occur, the enzyme selectivity was determined. The selectivity is defined quantitatively as the relative rate of hydrolysis of each

  7. Secretory IgA, IgG, and IgM immunoglobulins isolated simultaneously from colostral whey by selective thiophilic adsorption.

    Science.gov (United States)

    Hutchens, T W; Magnuson, J S; Yip, T T

    1990-03-27

    In order to evaluate the collective contribution of immunoglobulins to gastrointestinal and immune system development in newborn infants, it would be advantageous to develop a simple and rapid procedure for the selective and quantitative removal of all immunoglobulin classes from colostrum and milk. Toward this end, the major immunoglobulin classes typically present in colostrum (IgM, sIgA, and IgG) have been isolated simultaneously by selective removal from a porcine colostral whey model system using a single-step, preparative scale procedure termed thiophilic adsorption. At neutral pH, the salt-promoted thiophilic affinity of immunoglobulins for the synthetic sulfone-thioether ligands exploits a common (as yet unknown) structural feature of immunoglobulins. The adsorption and recovery procedures operate efficiently under mild buffer conditions permitting the subsequent comparative biochemical analyses of individual immunoglobulin classes for structural and functional heterogeneity. Thus, 1 liter columns of thiophilic adsorbent (T-gel) were employed to obtain pure, structurally intact immunoglobulins from 1 liter batches of porcine colostral whey. The identity and purity of the isolated immunoglobulins were determined under both structure-stabilizing ('native') conditions and denaturing conditions using high-performance size-exclusion chromatography, sucrose density gradient centrifugation, immunodiffusion techniques, SDS-polyacrylamide gradient gel electrophoresis with immunoblotting identification procedures, and two-dimensional electrophoresis. This is the first procedure known to us that allows for the simultaneous one-step isolation (under homologous conditions) of various immunoglobulins with preserved quaternary structure and apparent biological activity.

  8. Characterization of Neisseria cinerea, a nonpathogenic species isolated on Martin-Lewis medium selective for pathogenic Neisseria spp.

    OpenAIRE

    Knapp, J S; Totten, P A; Mulks, M H; Minshew, B H

    1984-01-01

    An asaccharolytic, gram-negative, oxidase-positive diplococcus was isolated on Martin-Lewis medium from the cervix of a patient attending an arthritis clinic at Seattle Public Health Hospital, Seattle, Wash. This strain, NRL 32165, did not produce detectable acid from glucose, maltose, sucrose, fructose, mannitol, or lactose in either cystine Trypticase agar (BBL Microbiology Systems, Cockeysville, Md.) or modified oxidation-fermentation medium and was identified presumptively as a glucose-ne...

  9. Rapid and selective extraction, isolation, preconcentration, and quantitation of small RNAs from cell lysate using on-chip isotachophoresis.

    Science.gov (United States)

    Schoch, Reto B; Ronaghi, Mostafa; Santiago, Juan G

    2009-08-07

    We present a technique which enables the separation of small RNAs-such as microRNAs (miRNAs), short interfering RNAs (siRNAs), and Piwi-interacting RNAs (piRNAs)-from >or=66 nucleotide RNAs and other biomolecules contained in a cell lysate. In particular, the method achieves separation of small RNAs from precursor miRNAs (pre-miRNAs) in less than 3 min. We use on-chip isotachophoresis (ITP) for the simultaneous extraction, isolation, preconcentration and quantitation of small RNAs (approximately 22 nucleotides) and employ the high-efficiency sieving matrix Pluronic F-127; a thermo-responsive triblock copolymer which allows convenient microchannel loading at low temperature. We present the isolation of small RNAs from the lysate of 293A human kidney cells, and quantitate the number of short RNA molecules per cell to be 2.9x10(7). We estimate this quantity is an aggregate of roughly 500 types of short RNA molecules per 293A cell. Currently, the minimal cell number for small RNA extraction and detection with our method is approximately 900 (from a 5 microL sample volume), and we believe that small RNA analysis from tens of cells is realizable. Techniques for rapid and sensitive extraction and isolation of small RNAs from cell lysate are much-needed to further uncover their full range and functionality, including RNA interference studies.

  10. Selection and Test of L-histidine Decarboxylase Enzyme Activity of Six Isolates of Histamine Forming Bacteria

    Directory of Open Access Journals (Sweden)

    Romauli Aya Sophia

    2007-11-01

    Full Text Available Six isolates of histamine forming bacteria were screened to see the degree of ability in producing histamine on modified Niven's medium. The result showed that the six bacteria were able to produce histamine by giving a pinkish color on the medium, which could be used as a preliminary identification of histamine-forming bacteria (HFB. The isolates were grown in liquid modified Niven medium to measure the production of histamine. The histamine produced were determined by Hardy and Smith method. The result showed that all of the isolates produced high level of histamine (92.35 - 305.49 mg/100 ml of the medium. From all of them, Enterobacter spp. produced the highest level of histamine (305.49 mg/100 ml. A synthetic medium was used to measure the growth pattern and optimum time required by Enterobacter spp and Morganella morganii (as control bacteria to produce the L-histidine decarboxylase enzyme (HDC which is responsible for histamine production. The result showed that for both bacteria, the optimum enzim production was 8 hours after incubation.

  11. Prevalence of Pathogenic Bacteria Isolated from Surgical Site and Wound Infection among Patients Admitted in some selected Hospitals in Sokoto Metropolis, Nigeria

    Directory of Open Access Journals (Sweden)

    UK Muhammad

    2014-09-01

    Full Text Available Surgical and open wounds are commonly encountered in clinical practice. This study was aim to determine the prevalence of pathogenic bacteria in surgical and open wound infection among patients admitted in some selected hospitals in Sokoto metropolis. A total of one hundred and fifty one (151 isolates were obtained from two hundred (200 surgical site and wound samples collected from patients in this study. The result showed that Usmanu Danfodiyo Teaching Hospital Sokoto (UDUTH had the highest number of clinical isolates with 64 gram positive and gram negative bacteria followed by Specialist Hospital Sokoto (S.H.S with 57 gram positive and gram negative bacteria and then Maryam Abacha Women and Children Hospital (MAWCH with 30 gram positive and gram negative bacteria. Gram positive cocci 108 (71.5% were more predominant pathogen isolated in the hospitals than gram negative bacilli 43 (28.5%. Staphylococcus aureus had the highest number of occurrence with 54(35.76% followed by Coagulate negative Staphylococci with 47(31.1% while Citrobacter freundii had the lowest number of occurrence with 2(1.32% isolates. Also, the susceptibility of the isolates to antimicrobial agents were carried out using Amoxacillin, Ampicillin, Erythromycin, Chloramphenicol, Ampiclox, Ciprofloxacin, Gentamycin, Tetracycline, Pefloxacin and Cotrimoxazole. The mean zone of inhibition recorded against Staphlococcus aureus by using Amoxacillin antibiotic is 2.20mm while with Citrobacter freundii is 1.00. DOI: http://dx.doi.org/10.3126/ije.v3i3.11066 International Journal of Environment Vol.3(3 2014: 89-103

  12. Resistance to erucic acid as a selectable marker for peroxisomal activity: isolation of revertants of an infantile Refsum disease cell line.

    Science.gov (United States)

    Bachir Bioukar, E; Straehli, F; Ng, K H; Rolland, M O; Hashimoto, T; Carreau, J P; Deschatrette, J

    1994-01-01

    A system based on the ability of cells to oxidize very long-chain fatty acids (VLCFA) was developed to select in vitro normal human fibroblasts from fibroblasts of patients suffering from peroxisomal disorders with multienzymatic deficiencies: Zellweger syndrome, neonatal adrenoleukodystrophy, infantile Refsum disease (IRD). Cells treated with various concentrations of erucic acid (C22:1 n-9) revealed an enhanced toxicity of this fatty acid for the fibroblasts of patients compared with normal cells. This differential toxicity is correlated with variable accumulations of C22:1 n-9 and the absence of beta-oxidation products in the mutants. Revertants from clonal IRD cell lines were isolated in the selective medium at frequencies ranging from 3 x 10(-7) to 4 x 10(-6) depending on the line. After six weeks of growth in the absence of selective pressure, the variants exhibited a resistance level to C22:1 n-9 identical to that of normal cells. Furthermore, beta-oxidation of VLCFA is re-established in these selected cells as well as dihydroxyacetone phosphate acyltransferase activity. Immunoblot experiments also demonstrated a restored pattern of acyl-CoA oxidase molecular forms. Last, immunofluorescence studies revealed the presence of cytoplasmic structures that were absent in the original IRD cells. Thus, both the deficiencies in metabolic pathways and paucity of the organelle are at least partially corrected in the selected clones.

  13. Development of a qPCR Strategy to Select Bean Genes Involved in Plant Defense Response and Regulated by the Trichoderma velutinum - Rhizoctonia solani Interaction.

    Science.gov (United States)

    Mayo, Sara; Cominelli, Eleonora; Sparvoli, Francesca; González-López, Oscar; Rodríguez-González, Alvaro; Gutiérrez, Santiago; Casquero, Pedro A

    2016-01-01

    Bean production is affected by a wide diversity of fungal pathogens, among them Rhizoctonia solani is one of the most important. A strategy to control bean infectious diseases, mainly those caused by fungi, is based on the use of biocontrol agents (BCAs) that can reduce the negative effects of plant pathogens and also can promote positive responses in the plant. Trichoderma is a fungal genus that is able to induce the expression of genes involved in plant defense response and also to promote plant growth, root development and nutrient uptake. In this article, a strategy that combines in silico analysis and real time PCR to detect additional bean defense-related genes, regulated by the presence of Trichoderma velutinum and/or R. solani has been applied. Based in this strategy, from the 48 bean genes initially analyzed, 14 were selected, and only WRKY33, CH5b and hGS showed an up-regulatory response in the presence of T. velutinum. The other genes were or not affected (OSM34) or down-regulated by the presence of this fungus. R. solani infection resulted in a down-regulation of most of the genes analyzed, except PR1, OSM34 and CNGC2 that were not affected, and the presence of both, T. velutinum and R. solani, up-regulates hGS and down-regulates all the other genes analyzed, except CH5b which was not significantly affected. As conclusion, the strategy described in the present work has been shown to be effective to detect genes involved in plant defense, which respond to the presence of a BCA or to a pathogen and also to the presence of both. The selected genes show significant homology with previously described plant defense genes and they are expressed in bean leaves of plants treated with T. velutinum and/or infected with R. solani.

  14. Development of a qPCR strategy to select bean genes involved in plant defence response and regulated by the Trichoderma velutinum - Rhizoctonia solani interaction

    Directory of Open Access Journals (Sweden)

    Sara Mayo

    2016-08-01

    Full Text Available Bean production is affected by a wide diversity of fungal pathogens, among them Rhizoctonia solani is one of the most important. A strategy to control bean infectious diseases, mainly those caused by fungi, is based on the use of biocontrol agents that can reduce the negative effects of plant pathogens and also can promote positive responses in the plant. Trichoderma is a fungal genus that is able to induce the expression of genes involved in plant defence response and also to promote plant growth, root development and nutrient uptake. In this article, a strategy that combines in silico analysis and real time PCR to detect additional bean defence-related genes, regulated by the presence of Trichoderma velutinum and/or R. solani has been applied. Based in this strategy, from From the 48 bean genes initially analysed, 14 were selected, and only WRKY33, CH5b and hGS showed an up-regulatory response in the presence of T. velutinum. The other genes were or not affected (OSM34 or down-regulated by the presence of this fungus. R. solani infection resulted in a down-regulation of most of the genes analyzed, except PR1, OSM34 and CNGC2 that were not affected, and the presence of both, T. velutinum and R. solani, up-regulates hGS and down-regulates all the other genes analyzed, except CH5b which was not significantly affected.As conclusion, the strategy described in the present work has been shown to be effective to detect genes involved in plant defence, which respond to the presence of a biocontrol agent or to a pathogen and also to the presence of both. The selected genes show significant homology with previously described plant defence genes and they are expressed in bean leaves of plants treated with T. velutinum and/or infected with R. solani.

  15. Development of a qPCR Strategy to Select Bean Genes Involved in Plant Defense Response and Regulated by the Trichoderma velutinum – Rhizoctonia solani Interaction

    Science.gov (United States)

    Mayo, Sara; Cominelli, Eleonora; Sparvoli, Francesca; González-López, Oscar; Rodríguez-González, Alvaro; Gutiérrez, Santiago; Casquero, Pedro A.

    2016-01-01

    Bean production is affected by a wide diversity of fungal pathogens, among them Rhizoctonia solani is one of the most important. A strategy to control bean infectious diseases, mainly those caused by fungi, is based on the use of biocontrol agents (BCAs) that can reduce the negative effects of plant pathogens and also can promote positive responses in the plant. Trichoderma is a fungal genus that is able to induce the expression of genes involved in plant defense response and also to promote plant growth, root development and nutrient uptake. In this article, a strategy that combines in silico analysis and real time PCR to detect additional bean defense-related genes, regulated by the presence of Trichoderma velutinum and/or R. solani has been applied. Based in this strategy, from the 48 bean genes initially analyzed, 14 were selected, and only WRKY33, CH5b and hGS showed an up-regulatory response in the presence of T. velutinum. The other genes were or not affected (OSM34) or down-regulated by the presence of this fungus. R. solani infection resulted in a down-regulation of most of the genes analyzed, except PR1, OSM34 and CNGC2 that were not affected, and the presence of both, T. velutinum and R. solani, up-regulates hGS and down-regulates all the other genes analyzed, except CH5b which was not significantly affected. As conclusion, the strategy described in the present work has been shown to be effective to detect genes involved in plant defense, which respond to the presence of a BCA or to a pathogen and also to the presence of both. The selected genes show significant homology with previously described plant defense genes and they are expressed in bean leaves of plants treated with T. velutinum and/or infected with R. solani. PMID:27540382

  16. The proteome of exudates from germinating Lupinus albus seeds is secreted through a selective dual-step process and contains proteins involved in plant defence.

    Science.gov (United States)

    Scarafoni, Alessio; Ronchi, Alessandro; Prinsi, Bhakti; Espen, Luca; Assante, Gemma; Venturini, Giovanni; Duranti, Marcello

    2013-03-01

    The general knowledge of defence activity during the first steps of seed germination is still largely incomplete. The present study focused on the proteins released in the exudates of germinating white lupin seeds. During the first 24 h, a release of proteins was observed. Initially (i.e. during the first 12 h), the proteins found in exudates reflected the composition of the seed, indicating a passive extrusion of pre-formed proteins. Subsequently, when the rate of protein release was at its highest, the composition of the released proteome changed drastically. This transition occurred in a short time, indicating that more selective and regulated events, such as secretory processes, took place soon after the onset of germination. The present study considered: (a) the characterization of the proteome accumulated in the germinating medium collected after the appearance of the post-extrusion events; (b) the biosynthetic origin and the modalities that are the basis of protein release outside the seeds; and (c) an assessment of antifungal activity of these exudates. The most represented protein in the exudate was chitinase, which was synthesized de novo. The other proteins are involved in the cellular mechanisms responding to stress events, including biotic ones. This exudate was effectively able to inhibit fungal growth. The results of the present study indicate that seed exudation is a dual-step process that leads to the secretion of selected proteins and thus is not a result of passive leakage. The released proteome is involved in protecting the spermosphere environment and thus may act as first defence against pathogens.

  17. Hypercapnic vasodilatation in isolated rat basilar arteries is exerted via low pH and does not involve nitric oxide synthase stimulation or cyclic GMP production

    DEFF Research Database (Denmark)

    You, J P; Wang, Qian; Zhang, W

    1994-01-01

    The relaxant effect of hypercapnia (15% CO2) was studied in isolated circular segments of rat basilar arteries with intact endothelium. The nitric oxide synthase inhibitor nitro-L-arginine (L-NOARG) and the cytosolic guanylate cyclase inhibitor methylene blue (MB), significantly reduced...

  18. Mutation in the S-ribosylhomocysteinase (luxS) gene involved in quorum sensing affects biofilm formation and virulence in a clinical isolate of Aeromonas hydrophila

    Science.gov (United States)

    A diarrheal isolate SSU of Aeromonas hydrophila produces a cytotoxic enterotoxin (Act) with cytotoxic, enterotoxic, and hemolytic activities. Our laboratory has characterized from the above Aeromonas strain, in addition to Act, the type 3- and T6-secretion systems and their effec...

  19. A genome-wide search for genes involved in type 2 diabetes in a recently genetically isolated population from the Netherlands

    NARCIS (Netherlands)

    Y.S. Aulchenko (Yurii); N. Vaessen (Norbert); P. Heutink (Peter); J. Pullen (Jan); P.J.L.M. Snijders (Pieter); A. Hofman (Albert); L.A. Sandkuijl (Lodewijk); J.J. Houwing-Duistermaat (Jeanine); S. Bennett (Simon); B.A. Oostra (Ben); C.M. van Duijn (Cock); M. Edwards (Mark)

    2003-01-01

    textabstractMultiple genes, interacting with the environment, contribute to the susceptibility to type 2 diabetes. We performed a genome-wide search to localize type 2 diabetes susceptibility genes in a recently genetically isolated population in the Netherlands. We identified 79 nuclear families wi

  20. Fermentation of Apple Juice with a Selected Yeast Strain Isolated from the Fermented Foods of Himalayan Regions and Its Organoleptic Properties

    Science.gov (United States)

    Kanwar, S. S.; Keshani

    2016-01-01

    Twenty-three Saccharomyces cerevisiae strains isolated from different fermented foods of Western Himalayas have been studied for strain level and functional diversity in our department. Among these 23 strains, 10 S. cerevisiae strains on the basis of variation in their brewing traits were selected to study their organoleptic effect at gene level by targeting ATF1 gene, which is responsible for ester synthesis during fermentation. Significant variation was observed in ATF1 gene sequences, suggesting differences in aroma and flavor of their brewing products. Apple is a predominant fruit in Himachal Pradesh and apple cider is one of the most popular drinks all around the world hence, it was chosen for sensory evaluation of six selected yeast strains. Organoleptic studies and sensory analysis suggested Sc21 and Sc01 as best indigenous strains for soft and hard cider, respectively, indicating their potential in enriching the local products with enhanced quality. PMID:27446050

  1. Utilization of Multi-Immunization and Multiple Selection Strategies for Isolation of Hapten-Specific Antibodies from Recombinant Antibody Phage Display Libraries

    Directory of Open Access Journals (Sweden)

    Antti Tullila

    2017-05-01

    Full Text Available Phage display technology provides a powerful tool for the development of novel recombinant antibodies. In this work, we optimized and streamlined the recombinant antibody discovery process for haptens as an example. A multi-immunization approach was used in order to avoid the need for construction of multiple antibody libraries. Selection methods were developed to utilize the full potential of the recombinant antibody library by applying four different elution conditions simultaneously. High-throughput immunoassays were used to analyse the binding properties of the individual antibody clones. Different carrier proteins were used in the immunization, selection, and screening phases to avoid enrichment of the antibodies for the carrier protein epitopes. Novel recombinant antibodies against mycophenolic acid and ochratoxin A, with affinities up to 39 nM and 34 nM, respectively, were isolated from a multi-immunized fragment antigen-binding (Fab library.

  2. Utilization of Multi-Immunization and Multiple Selection Strategies for Isolation of Hapten-Specific Antibodies from Recombinant Antibody Phage Display Libraries

    Science.gov (United States)

    Tullila, Antti; Nevanen, Tarja K.

    2017-01-01

    Phage display technology provides a powerful tool for the development of novel recombinant antibodies. In this work, we optimized and streamlined the recombinant antibody discovery process for haptens as an example. A multi-immunization approach was used in order to avoid the need for construction of multiple antibody libraries. Selection methods were developed to utilize the full potential of the recombinant antibody library by applying four different elution conditions simultaneously. High-throughput immunoassays were used to analyse the binding properties of the individual antibody clones. Different carrier proteins were used in the immunization, selection, and screening phases to avoid enrichment of the antibodies for the carrier protein epitopes. Novel recombinant antibodies against mycophenolic acid and ochratoxin A, with affinities up to 39 nM and 34 nM, respectively, were isolated from a multi-immunized fragment antigen-binding (Fab) library. PMID:28561803

  3. Interaction of activity involvement and recreational location selection behavior in Lowland City: A case study of public parks in Saga City, Japan

    Institute of Scientific and Technical Information of China (English)

    IAMTRAKUL Pawinee; TEKNOMO Kardi; GE Jian; HOKAO Kazunori

    2005-01-01

    Public parks provide many benefits to the community as the representatives of green area. The allocation of public places plays an extremely important role in the daily lives of inhabitants especially for recreational use that could enhance the quality of life of residents in the vicinity. To understand park users' behavior is one of the most important prerequisites for assessing the participation in public service from the park users' point of view. The pattern of park utilization on location and activity selection are important elements in behavioral study, while the public parks topograph may also influence the typical user's behavior. Questionnaire survey on park utilization was used to investigate the interaction between activity involvement and recreational location with the use of linear discriminant analysis (LDA) model. The study found that public park users' behavior is influenced not only by social characteristics but also by the recreational activities and their specific location characteristics. We found that about 45 percent of park visitors are local residents living within a radius of 3 km preferred travel to parks near their residential area. This implies that location selection behavior is correlated with travel distance, travel time and travel cost. Visit frequencies and on site expenditures reflect the recreation behavior for different type of activities. The overall information can be usefully applied by decision makers to launch appropriate public policy in consistence with the useful results of this study.

  4. Involvement of HDAC1 and HDAC3 in the Pathology of Polyglutamine Disorders: Therapeutic Implications for Selective HDAC1/HDAC3 Inhibitors

    Directory of Open Access Journals (Sweden)

    Elizabeth A. Thomas

    2014-05-01

    Full Text Available Histone deacetylases (HDACs enzymes, which affect the acetylation status of histones and other important cellular proteins, have been recognized as potentially useful therapeutic targets for a broad range of human disorders. Emerging studies have demonstrated that different types of HDAC inhibitors show beneficial effects in various experimental models of neurological disorders. HDAC enzymes comprise a large family of proteins, with18 HDAC enzymes currently identified in humans. Hence, an important question for HDAC inhibitor therapeutics is which HDAC enzyme(s is/are important for the amelioration of disease phenotypes, as it has become clear that individual HDAC enzymes play different biological roles in the brain. This review will discuss evidence supporting the involvement of HDAC1 and HDAC3 in polyglutamine disorders, including Huntington’s disease, and the use of HDAC1- and HDAC3-selective HDAC inhibitors as therapeutic intervention for these disorders. Further, while HDAC inhibitors are known alter chromatin structure resulting in changes in gene transcription, understanding the exact mechanisms responsible for the preclinical efficacy of these compounds remains a challenge. The potential chromatin-related and non-chromatin-related mechanisms of action of selective HDAC inhibitors will also be discussed.

  5. Equilibrium studies in solution involving nickel(II) complexes of flexidentate Schiff base ligands: isolation and structural characterization of the planar red and octahedral green species involved in the equilibrium.

    Science.gov (United States)

    Mukhopadhyay, Suman; Mandal, Debdas; Ghosh, Dipesh; Goldberg, Israel; Chaudhury, Muktimoy

    2003-12-15

    Three new flexidentate 5-substituted salicylaldimino Schiff base ligands (L1-OH-L3-OH) based on 1-(2-aminoethyl)piperazine (X=H, L1-OH; X=NO2, L2-OH; and X=Br, L3-OH) and their nickel(II) complexes (1a, 1b, 2, and 3) have been reported. The piperazinyl arm of these ligands can in principle have both boat and chair conformations that allow the ligands to bind the Ni(II) center in an ambidentate manner, forming square-planar and/or octahedral complexes. The nature of substitution in the salicylaldehyde aromatic ring and the type of associated anion in the complexes have profound influences on the coordination geometry of the isolated products. With the parent ligand L1-OH, the product obtained is either a planar red compound [Ni(L1-O)]+, isolated as tetraphenylborate salt (1a), or an octahedral green compound [Ni(L1-NH)(H2O)3](2+), isolated with sulfate anion (1b); both have been crystallographically characterized. In aqueous solution, both these planar (S=0) and octahedral (S=1) forms are in equilibrium that has been followed in the temperature range 298-338 K by 1H NMR technique using the protocol of Evans's method. The large exothermicity of the equilibrium process [Ni(L1-O)]+ + 3H2O + H+[Ni(L1-NH)(H2O)3](2+) (DeltaH degrees=-46 +/- 0.2 kJ mol(-1) and DeltaS degrees=-133 +/- 5 J K(-1) mol(-1)) reflects formation of three new Ni-OH2 bonds in going from planar to the octahedral species. With the 5-nitro derivative ligand L2-OH, the sole product is an octahedral compound 2, isolated as a sulfate salt while with the bromo derivative ligand L3-OH, the exclusive product is a planar molecule 3 with associated tetraphenylborate anion. Both 2 and 3 have been structurally characterized by X-ray diffraction analysis.

  6. Improvement of mannitol lysine crystal violet brilliant green agar for the selective isolation of H2S-positive Salmonella.

    Science.gov (United States)

    Kodaka, H; Mizuochi, S; Honda, T; Yamaguchi, K

    2000-12-01

    Mannitol lysine crystal violet brilliant green agar (MLCB) is widely used in Japan for Salmonella isolation because the medium has been commercially available. Colonies of Salmonella on MLCB appear colorless with black centers due to H2S gas production; however, most Citrobacter freundii also produce H2S gas. In order to distinguish H2S-positive Salmonella from C. freundii we have improved MLCB. To MLCB was added 1% lactose (L-MLCB). The relation for pH and black center colony formation was examined. The pH of MLCB and L-MLCB inoculated with Salmonella species was slightly acid after 7 h, but the pH of L-MLCB inoculated with C. freundii did not become acid for 24 h. The colony of C. freundii did not have a black center because the production of acid from lactose lowers the pH below 10 where it is needed for H2S to react with iron to produce black pigments. Of 99 Salmonella strains including 13 serotypes tested, all strains had the same colony morphologies on MLCB and L-MLCB. When MLCB and L-MLCB were evaluated with 36 C. freundii strains isolated from foods, only colonies on MLCB had black centers. We conclude that L-MLCB is useful for detection of nonlactose-fermenting, H2S-positive Salmonella in food samples.

  7. Selection of potential Enterococcus faecium isolated from Thai native chicken for probiotic use according to the in vitro properties

    Directory of Open Access Journals (Sweden)

    Napaporn Lertworapreecha

    2011-02-01

    Full Text Available Sixty strains of E. faecium were isolated from 30 samples of native chickens’ gastrointestinal tracts. All strains weretested on acid and bile tolerance. Fifteen strains passed the acid tolerance test. The best five strains were EFMC 17, 21 and24; EFMD 25; EFMI 47 and 49. Only four strains, EFMC 21; EFMD 30; EFMI 47, and 49, survived 4 hours of bile exposure.Fifteen strains that passed the acid tolerance test were tested for their ability of intestinal mucus attachment. The resultsindicated that all strains were able to attach to intestinal mucus. For the ability of pathogenic bacteria inhibition test, theresult found seven strains (EFMC 17, 21 and 24; EFMD 29 and 30; EFMI 46 and 49 showed better performance than strainEFC. All seven strains were acid producer, but only four strains (EFMC 21; EFMD 25; EFMI 47 and 49 were able to releasebacteriocin. Based on proper probiotic properties two strains (EFMI 47 and 49 of E. faecium isolated from Thai native chicksin this study have a potential use as probiotics. Antimicrobial susceptibility test of these two strains have been also performed;they were susceptible to amoxicillin/clavulanic, ciprofloxacin, gentamycin, trimethoprime/sulphamethoxazole, vancomycin,and trimethoprim. On the other hand, they were resistant to cefotaxime, erythromycin, and tetracycline. The DNA-DNAhybridization percentage of DNA-DNA homology to E. faecium NRIC 1145 of EFMI 47 and EFMI 49 were 82.36 and 78.63%,respectively.

  8. [Selected properties of lactose-fermenting and non-fermenting Salmonella agona strains isolated from specimens from hospitalized infants].

    Science.gov (United States)

    Kukulska, D; Zimmer, A

    1989-01-01

    The aim of this study was to compare some of the properties of 28 lactose-positive and 28 lactose-negative Salmonella agona strains isolated from faeces of infants hospitalized in the same hospital. Some of biochemical properties, sensitivity to 14 antibiotics and chemotherapeutic agents and sensitivity to bacteriophages used for typing of this Salmonella genus were tested. Results of biochemical examinations revealed that lactose-fermenting strains retain the remaining of Salmonella of subspecies I. Two biochemical features are of particular importance: the ability to ferment lactose on all lactose containing media and a lack of the ability to produce H2S on Kligler medium. These two features differentiate lactose-fermenting strains of Salmonella from non-lactose fermenting ones. Antibiotic sensitivity pattern differed between lactose-positive and lactose-negative strains. Lactose-positive strains showed higher degree of resistance than lactose-negative strains. The differences in resistance were seen in the case of chloramphenicol, doxycycline, gentamicin and tetracycline. Both lactose-positive and lactose-negative strains were sensitive to colistin, neomycin, nitrofurantoin and nalidixic acid. They were resistant to ampicillin, cloxacillin, rifampicin, streptomycin, sulfatiazol and biseptol. Bacteriophage typing revealed that all lactose-negative strains isolated in this study from clinical samples belonged to the same phage pattern V. Lactose-positive strains belonged to two phage types VB and XI. Type VB prevailed.

  9. Mimicking the germinal center reaction in hybridoma cells to isolate temperature-selective anti-PEG antibodies.

    Science.gov (United States)

    Su, Yu-Cheng; Al-Qaisi, Talal S; Tung, Hsin-Yi; Cheng, Tian-Lu; Chuang, Kuo-Hsiang; Chen, Bing-Mae; Roffler, Steve R

    2014-01-01

    Modification of antibody class and binding properties typically requires cloning of antibody genes, antibody library construction, phage or yeast display and recombinant antibody expression. Here, we describe an alternative "cloning-free" approach to generate antibodies with altered antigen-binding and heavy chain isotype by mimicking the germinal center reaction in antibody-secreting hybridoma cells. This was accomplished by lentiviral transduction and controllable expression of activation-induced cytidine deaminase (AID) to generate somatic hypermutation and class switch recombination in antibody genes coupled with high-throughput fluorescence-activated cell sorting (FACS) of hybridoma cells to detect altered antibody binding properties. Starting from a single established hybridoma clone, we isolated mutated antibodies that bind to a low-temperature structure of polyethylene glycol (PEG), a polymer widely used in nanotechnology, biotechnology and pharmaceuticals. FACS of AID-infected hybridoma cells also facilitated rapid identification of class switched variants of monoclonal IgM to monoclonal IgG. Mimicking the germinal center reaction in hybridoma cells may offer a general method to identify and isolate antibodies with altered binding properties and class-switched heavy chains without the need to carry out DNA library construction, antibody engineering and recombinant protein expression.

  10. Confirmation of Isolated Cu2+ Ions in SSZ-13 Zeolite as Active Sites in NH3-Selective Catalytic Reduction

    NARCIS (Netherlands)

    Deka, U.; Juhin, A.F.; Eilertsen, E.A.; Emerich, H.; Green, M.A.; Korhonen, S.T.; Weckhuysen, B.M.; Beale, A.M.

    2012-01-01

    NH3-Selective Catalytic Reduction (NH3-SCR) is a widely used technology for NOx reduction in the emission control systems of heavy duty diesel vehicles. Copper-based ion exchanged zeolites and in particular Cu-SSZ-13 (CHA framework) catalysts show both exceptional activity and hydrothermal stability

  11. Synergistic Effects of n-Hexane Fraction of Parkia biglobosa (Jacq. Bark Extract and Selected Antibiotics on Bacterial Isolates

    Directory of Open Access Journals (Sweden)

    Oluwatayo E. Abioye

    2017-02-01

    Full Text Available The incidence of resistance to commonly used antimicrobial agents by microbial pathogens demands increased effort in the development of effective ways of treating infections and diseases. The n-hexane fraction of lyophilized crude bark extract of Parkia biglobosa (Jacq. was prepared and, in combination with selected antibiotics, assayed for antimicrobial activity against some selected bacterial pathogens using time-kill assay. Protein leakage analysis of the combined agents was performed using Bradford protein quantification method. Determination of active compounds present in the n-hexane fraction was done using Fourier Transform Infrared Spectroscopy (FTIR. While time-kill assay detected 43.33% synergy; 56.67% indifference and no antagonism at 1/2 × minimum inhibitory concentration (MIC, 1 × MIC exhibited 55% synergy, 45% indifference and no antagonism. Protein leakages from the cells of selected bacteria ranged from 1.20 µg/mL to 256.93 µg/mL. The presence of a phenyl group, an aromatic ring and phenolic compounds in the n-hexane fraction was confirmed at 2162 cm−1–2020 cm−1, 1605 cm−1–1533 cm−1 and 1438 cm−1–1444 cm−1 spectra peaks, respectively. The observed antibiotic−n-hexane fraction synergistic interaction revealed the improved antibacterial activity of the selected antibiotics. Hence, exploration of a combination of antibiotics with plant secondary metabolites is hereby advocated in the global quest for means of combating infectious diseases caused by multidrug-resistant pathogens.

  12. Characterization and selection of autochthonous lactic acid bacteria isolated from traditional Iberian dry-fermented salchichón and chorizo sausages.

    Science.gov (United States)

    Benito, M J; Martín, A; Aranda, E; Pérez-Nevado, F; Ruiz-Moyano, S; Córdoba, M G

    2007-08-01

    A total of 192 lactic acid bacteria were isolated from 2 types of naturally fermented dry sausages at 3 different stages of the ripening process in order to select the most suitable strains as starter cultures in dry-cured sausage manufacture according to their technological characteristics such as glucose fermentation, lactic and acetic acid production, and proteolytic, lipolytic, and antimicrobial activities. Identification of the isolates revealed that 31.2% were Pediococcus pentosaceus, 26.9% Lactococcus lactis, 18.6% Pediococcus acidilactici, 17% Lactobacillus brevis, and sporadic isolates of Leuconostoc mesenteroides, Lactobacillus plantarum, and Lactobacillus curvatus. Most of the strains did not produce gas from glucose and showed the capacity to produce lactic acid rapidly. Some 25% of the strains were able to degrade tributyrin (esterase activity), but none showed lipolytic activity against olive oil and pork fat. Only 3 strains of P. acidilactici showed weak proteolytic activity against myofibrillar or sarcoplasmic proteins. Also, the same strains showed antimicrobial activity against Listeria monocytogenes. Nine strains with the best properties were preselected and tested for biogenic amine production. The results showed that two of the strains, identified as P. acidilactici by polymerase chain reaction, had the potential to be further tested as starter cultures in pilot processing of Iberian sausages.

  13. Rapid Isolation of the Trichoderma Strain with Higher Degrading Ability of a Filter Paper and Superior Proliferation Characteristics Using Avicel Plates and the Double-Layer Selection Medium

    Science.gov (United States)

    Toyama, Hideo; Nakano, Megumi; Satake, Yuuki; Toyama, Nobuo

    The cost of cellulase is still a problem for bioethanol production. As the cellulase of Trichoderma reesei is applicable for producing ethanol from cellulosic materials, the cellulase productivity of this fungus should be increased. Therefore, we attempted to develop a system to isolate the strain with higher degrading ability of a filter paper and superior proliferation characteristics among the conidia treated with the mitotic arrester, colchicine. When green mature conidia of T. reesei RUT C-30 were swollen, autopolyploidized, and incubated in the double-layer selection medium containing Avicel, colonies appeared on the surface earlier than the original strain. When such colonies and the original colony were incubated on the Avicel plates, strain B5, one of the colonies derived from the colchicinetreated conidia, showed superior proliferation characteristics. Moreover, when strain B5 and the original strain were compared in the filter paper degrading ability and the cellulose hydrolyzing activity, strain B5 was also superior to the original strain. It was suspected that superior proliferation characteristics of strain B5 reflects higher filter paper degrading ability. Thus, we concluded that the Trichoderma strain with higher degrading ability of a filter paper and superior proliferation characteristics can be isolated using Avicel plates and the double-layer selection medium.

  14. Selection of prophylactic antibiotics according to the microorganisms isolated from surgical site infections (SSIs) in a previous series of surgeries reduces SSI incidence after pancreaticoduodenectomy.

    Science.gov (United States)

    Kondo, Kazuhiro; Chijiiwa, Kazuo; Ohuchida, Jiro; Kai, Masahiro; Fujii, Yoshiro; Otani, Kazuhiro; Hiyoshi, Masahide; Nagano, Motoaki; Imamura, Naoya

    2013-03-01

    The incidence of surgical site infections (SSIs) is high after pancreaticoduodenectomy (PD). We divided 116 consecutive patients who underwent PD into an early group (n = 58) and a later group (n = 58) according to time of surgery. In both groups, endoscopic retrograde biliary drainage was mainly employed for the patients with obstructive jaundice. In the later group, prophylactic antibiotics were selected according to the susceptibility of microorganisms isolated from SSIs in the early group. The incidence of SSIs was compared between the groups. The background characteristics (including methods of preoperative biliary drainage and microorganisms in the bile obtained before or during operation) of the patients were not significantly different between the groups, except for the serum albumin level, which was lower in the later group than in the early group (P = 0.0026). The incidence of SSIs was significantly lower in the later group (24.1 %) than in the early group (46.6 %) (P = 0.0116). Belonging to the later group was one independent negative risk factor for SSI. Selection of prophylactic antibiotics on the basis of microorganisms isolated from SSIs in the early group contributed to the reduced incidence of SSIs in the later group after PD.

  15. Rapid isolation of the Trichoderma strain with higher degrading ability of a filter paper and superior proliferation characteristics using avicel plates and the double-layer selection medium.

    Science.gov (United States)

    Toyama, Hideo; Nakano, Megumi; Satake, Yuuki; Toyama, Nobuo

    2008-03-01

    The cost of cellulase is still a problem for bioethanol production. As the cellulase of Trichoderma reesei is applicable for producing ethanol from cellulosic materials, the cellulase productivity of this fungus should be increased. Therefore, we attempted to develop a system to isolate the strain with higher degrading ability of a filter paper and superior proliferation characteristics among the conidia treated with the mitotic arrester, colchicine. When green mature conidia of T. reesei RUT C-30 were swollen, autopolyploidized, and incubated in the double-layer selection medium containing Avicel, colonies appeared on the surface earlier than the original strain. When such colonies and the original colony were incubated on the Avicel plates, strain B5, one of the colonies derived from the colchicine-treated conidia, showed superior proliferation characteristics. Moreover, when strain B5 and the original strain were compared in the filter paper degrading ability and the cellulose hydrolyzing activity, strain B5 was also superior to the original strain. It was suspected that superior proliferation characteristics of strain B5 reflects higher filter paper degrading ability. Thus, we concluded that the Trichoderma strain with higher degrading ability of a filter paper and superior proliferation characteristics can be isolated using Avicel plates and the double-layer selection medium.

  16. Isolation and characterization of Bacillus subtilis genes involved in siderophore biosynthesis: relationship between B. subtilis sfpo and Escherichia coli entD genes.

    OpenAIRE

    Grossman, T H; Tuckman, M; Ellestad, S; Osburne, M S

    1993-01-01

    In response to iron deprivation, Bacillus subtilis secretes a catecholic siderophore, 2,3-dihydroxybenzoyl glycine, which is similar to the precursor of the Escherichia coli siderophore enterobactin. We isolated two sets of B. subtilis DNA sequences that complemented the mutations of several E. coli siderophore-deficient (ent) mutants with defective enterobactin biosynthesis enzymes. One set contained DNA sequences that complemented only an entD mutation. The second set contained DNA sequence...

  17. Selection and characterisation of lactic acid bacteria isolated from different origins for ensilingRobinia pseudoacacia and Morus alba L. leaves

    Institute of Scientific and Technical Information of China (English)

    NI Kui-kui; YANG Hui-xiao; HUA Wei; WANG Yan-ping; PANG Hui-li

    2016-01-01

    The objective of this study was to isolate lactic acid bacteria (LAB) strains from different origins and to select the best strains for ensilingRobinia pseudoacacia (RB) and Morus albaL. (MB)leaves. The LAB strains were inoculated into the extracted liquid obtained from RB and MB leaves to evaluate the fermentation products. 11 LAB strains were selected for further experiments based on the highest products of lactic or acetic acid, including 1 strain of Weissela confusa, 2 ofLactobacilus reuteri and 8 ofLactobacilus plantarum.The API 50 CH fermentation experiment indicated that al of the selected 11 LAB strains utilised most of the carbohydrates. Al the strains grew at temperatures between 10 and 45°C and at a pH of 3.5 to 4.5; however,L. reuteri F7 and F8 tolerated a pH as low as 3.0. Al 11 LAB strains showed antibacterial activity against Listeria monocytogens,Escherichia coil,Salmonelasp. and Acetobacter pasteurianus; however, after excluding the effect of organic acids, only F7 and F8 stil exhibited antibacterial activity. The present study indicated that the selected 11 LAB strains could be used to prepare silages of RB and MB leaves, especialyL. reuteri F7 and F8.

  18. Microarray-based comparative genomic profiling of reference strains and selected Canadian field isolates of Actinobacillus pleuropneumoniae

    Directory of Open Access Journals (Sweden)

    MacInnes Janet I

    2009-02-01

    Full Text Available Abstract Background Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia, is a highly contagious respiratory pathogen that causes severe losses to the swine industry worldwide. Current commercially-available vaccines are of limited value because they do not induce cross-serovar immunity and do not prevent development of the carrier state. Microarray-based comparative genomic hybridizations (M-CGH were used to estimate whole genomic diversity of representative Actinobacillus pleuropneumoniae strains. Our goal was to identify conserved genes, especially those predicted to encode outer membrane proteins and lipoproteins because of their potential for the development of more effective vaccines. Results Using hierarchical clustering, our M-CGH results showed that the majority of the genes in the genome of the serovar 5 A. pleuropneumoniae L20 strain were conserved in the reference strains of all 15 serovars and in representative field isolates. Fifty-eight conserved genes predicted to encode for outer membrane proteins or lipoproteins were identified. As well, there were several clusters of diverged or absent genes including those associated with capsule biosynthesis, toxin production as well as genes typically associated with mobile elements. Conclusion Although A. pleuropneumoniae strains are essentially clonal, M-CGH analysis of the reference strains of the fifteen serovars and representative field isolates revealed several classes of genes that were divergent or absent. Not surprisingly, these included genes associated with capsule biosynthesis as the capsule is associated with sero-specificity. Several of the conserved genes were identified as candidates for vaccine development, and we conclude that M-CGH is a valuable tool for reverse vaccinology.

  19. Selective mobilization of saturated fatty acids in isolated adipocytes of hibernating 13-lined ground squirrels Ictidomys tridecemlineatus.

    Science.gov (United States)

    Price, Edwin R; Armstrong, Christopher; Guglielmo, Christopher G; Staples, James F

    2013-01-01

    Fatty acids are not mobilized from adipocyte triacylglycerols uniformly but rather some are preferentially mobilized while others are preferentially retained. In many vertebrate species, the pattern of differential mobilization is determined by the physical and chemical properties of each fatty acid. Fatty acids with shorter chains and more double bonds tend to be more readily mobilized than others, a pattern observed both in whole-animal studies and in isolated adipocytes. Several hibernating species seem to break this pattern, however, and retain 18:2ω6 (linoleic acid) while mobilizing saturated fatty acids such as 18:0. We sought to confirm this pattern in adipocytes of a hibernator, the 13-lined ground squirrel Ictidomys tridecemlineatus, and to investigate mobilization patterns for the first time at hibernation temperature. We isolated adipocytes from summer active and winter torpid squirrels and incubated them with 1 μM norepinephrine at 4°C (7 h) and 37°C (90 min). We measured the proportion of each fatty acid in the adipose tissue and in the buffer at the end of incubation. Patterns of mobilization were similar in both seasons and incubation temperatures. Saturated fatty acids (18:0 and 16:0) were highly mobilized relative to the average, while some unsaturated fatty acids (notably, 18:1ω9 and 18:2ω6) were retained. We conclude that hibernators have unique mechanisms at the level of adipose tissue that preferentially mobilize saturated fatty acids. Additionally, we found that adipocytes from hibernating squirrels produced more glycerol than those from summer squirrels (regardless of temperature), indicating a higher lipolytic capacity in hibernating squirrels.

  20. Selective enrichment media bias the types of Salmonella enterica strains isolated from mixed strain cultures and complex enrichment broths.

    Science.gov (United States)

    Gorski, Lisa

    2012-01-01

    For foodborne outbreak investigations it can be difficult to isolate the relevant strain from food and/or environmental sources. If the sample is contaminated by more than one strain of the pathogen the relevant strain might be missed. In this study mixed cultures of Salmonella enterica were grown in one set of standard enrichment media to see if culture bias patterns emerged. Nineteen strains representing four serogroups and ten serotypes were compared in four-strain mixtures in Salmonella-only and in cattle fecal culture enrichment backgrounds using Salmonella enrichment media. One or more strain(s) emerged as dominant in each mixture. No serotype was most fit, but strains of serogroups C2 and E were more likely to dominate enrichment culture mixtures than strains of serogroups B or C1. Different versions of Rappaport-Vassiliadis (RV) medium gave different patterns of strain dominance in both Salmonella-only and fecal enrichment culture backgrounds. The fittest strains belonged to serogroups C1, C2, and E, and included strains of S. Infantis, S. Thompson S. Newport, S. 6,8:d:-, and S. Give. Strains of serogroup B, which included serotypes often seen in outbreaks such as S. Typhimurium, S. Saintpaul, and S. Schwarzengrund were less likely to emerge as dominant strains in the mixtures when using standard RV as part of the enrichment. Using a more nutrient-rich version of RV as part of the protocol led to a different pattern of strains emerging, however some were still present in very low numbers in the resulting population. These results indicate that outbreak investigations of food and/or other environmental samples should include multiple enrichment protocols to ensure isolation of target strains of Salmonella.

  1. Selected Essential Oils as Antifungal Agents Against Antibiotic-Resistant Candida spp.: In Vitro Study on Clinical and Food-Borne Isolates.

    Science.gov (United States)

    Rajkowska, Katarzyna; Kunicka-Styczyńska, Alina; Maroszyńska, Marta

    2017-01-01

    Candida spp. cause significant health problems, inducing various types of superficial and deep-seated mycoses in humans. As a result of the increasing antibiotic resistance among pathogenic yeasts, the interest in alternative agents of antifungal activity is growing. This study evaluated the antimicrobial activity of selected essential oils (EOs) against Candida clinical and food-borne strains, including antibiotic-resistant isolates, in relation to yeast cell surface hydrophobicity (CSH). Candida strains showed different range of susceptibility to tea tree, thyme, peppermint, and clove oils, and peppermint oil demonstrated the lowest anticandidal activity with minimal inhibitory concentrations (MICs) of 0.03-8.0% v/v. MIC values for thyme and clove oils ranged from 0.03% to 0.25% v/v, and for tea tree oil-from 0.12% to 2.0% v/v. The exception was Candida tropicalis food-borne strain, the growth of which was inhibited after application of EOs at concentration of 8% v/v. Due to diverse yeast susceptibility to EOs, isolates were divided into five clusters in a principal component analysis model, each containing both clinical and food-borne strains. Hydrophobic properties of yeast were also diversified, and 37% of clinical and 50% of food-borne strains exhibited high hydrophobicity. The study indicates high homology of clinical and food-borne Candida isolates in relation to their susceptibility to anticandidal agents and hydrophobic properties. The susceptibility of yeasts to EOs could be partially related to their CSH. High antifungal activity of examined EOs, also against antibiotic-resistant isolates, indicates their usefulness as agents preventing the development of Candida strains of different origin.

  2. Mutant-prevention concentration and mechanism of resistance in clinical isolates and enrofloxacin/marbofloxacin-selected mutants of Escherichia coli of canine origin.

    Science.gov (United States)

    Gebru, Elias; Choi, Myung-Jin; Lee, Seung-Jin; Damte, Dereje; Park, Seung Chun

    2011-10-01

    The antibacterial activity and selection of resistant bacteria, along with mechanisms of fluoroquinolone resistance, were investigated by integrating the static [MIC or mutant-prevention concentration (MPC)] and in vitro dynamic model approaches using Escherichia coli isolates from diseased dogs. Using the dynamic models, selected E. coli strains and enrofloxacin and marbofloxacin at a range of simulated area under concentration-time curve over a 24 h interval (AUC(24 h))/MIC ratios were investigated. Our results indicated increasing losses in susceptibility of E. coli upon continuous exposure to enrofloxacin and marbofloxacin in vitro. This effect was transferable to other fluoroquinolones, as well as to structurally unrelated drugs. Our results also confirmed an AUC(24 h)/MIC (AUC(24 h)/MPC)-dependent antibacterial activity and selection of resistant E. coli mutants, in which maximum losses in fluoroquinolone susceptibility occurred at simulated AUC(24 h)/MIC ratios of 40-60. AUC(24 h)/MPC ratios of 39 (enrofloxacin) and 32 (marbofloxacin) were considered protective against the selection of resistant mutants of E. coli. Integrating our MIC and MPC data with published pharmacokinetic information in dogs revealed a better effect of the conventional dosing regimen of marbofloxacin than that of enrofloxacin in restricting the selection of resistant mutants of E. coli. Target mutations, especially at codon 83 (serine to leucine) of gyrA, and overexpression of efflux pumps contributed to resistance development in both clinically resistant and in vitro-selected mutants of E. coli. We also report here a previously undescribed mutation at codon 116 of parC in two laboratory-derived resistant mutants of E. coli. Additional studies would determine the exact role of this mutation in fluoroquinolone susceptibility, as well as establish the importance of our findings in the clinical setting.

  3. Preparation of on-plate immobilized metal ion affinity chromatography platform via dopamine chemistry for highly selective isolation of phosphopeptides with matrix assisted laser desorption/ionization mass spectrometry analysis.

    Science.gov (United States)

    Shi, Chenyi; Lin, Qinrui; Deng, Chunhui

    2015-04-01

    In this study, a novel on-plate IMAC technique was developed for highly selective enrichment and isolation of phosphopeptides with high-throughput MALDI-TOF-MS analysis. At first, a MALDI plate was coated with polydopamine (PDA), and then Ti(4+) was immobilized on the PDA-coated plate. The obtained IMAC plate was successfully applied to the highly selective enrichment and isolation of phosphopeptides in protein digests and human serum. Because of no loss of samples, the on-plate IMAC platform exhibits excellent selectivity and sensitivity in the selective enrichment and isolation of phosphopeptides, which provides a potential technique for high selectivity in the detection of low-abundance phosphopeptides in biological samples.

  4. Selection of nitrogen-fixing deficient Burkholderia vietnamiensis strains by cystic fibrosis patients: involvement of nif gene deletions and auxotrophic mutations.

    Science.gov (United States)

    Menard, Aymeric; Monnez, Claire; Estrada de Los Santos, Paulina; Segonds, Christine; Caballero-Mellado, Jesus; Lipuma, John J; Chabanon, Gerard; Cournoyer, Benoit

    2007-05-01

    Burkholderia vietnamiensis is the third most prevalent species of the Burkholderia cepacia complex (Bcc) found in cystic fibrosis (CF) patients. Its ability at fixing nitrogen makes it one of the main Bcc species showing strong filiations with environmental reservoirs. In this study, 83% (29 over 35) of the B. vietnamiensis CF isolates and 100% of the environmental ones (over 29) were found expressing the dinitrogenase complex (encoded by the nif cluster) which is essential in N(2) fixation. Among the deficient strains, two were found growing with ammonium chloride suggesting that they were defective in N(2) fixation, and four with amino acids supplements suggesting that they were harbouring auxotrophic mutations. To get insights about the genetic events that led to the emergence of the N(2)-fixing defective strains, a genetic analysis of B. vietnamiensis nitrogen-fixing property was undertaken. A 40-kb-long nif cluster and nif regulatory genes were identified within the B. vietnamiensis strain G4 genome sequence, and analysed. Transposon mutagenesis and nifH genetic marker exchanges showed the nif cluster and several other genes like gltB (encoding a subunit of the glutamate synthase) to play a key role in B. vietnamiensis ability at growing in nitrogen-free media. nif cluster DNA probings of restricted genomic DNA blots showed a full deletion of the nif cluster for one of the N(2)-fixing defective strain while the other one showed a genetic organization similar to the one of the G4 strain. For 17% of B. vietnamiensis clinical strains, CF lungs appeared to have favoured the selection of mutations or deletions leading to N(2)-fixing deficiencies.

  5. Isolation and selection of P-dissolving microbes%解磷微生物的分离与筛选

    Institute of Scientific and Technical Information of China (English)

    钟传青; 曹广祥; 黄为一

    2014-01-01

    Phosphorus is the main component of organic compounds in plant and necessary for the biological nitrogen fixation.Most of the phosphorus is insoluble in soil,which cannot be absorbed by plants directly.P-dissolving microbes can promote the transformation of insoluble phosphorus to available phosphorus in soil.The study was centered on the current situation of effective phosphorus deficiency in cultivated land.Diversity of P-dissolving microbes was explored by construction of screening model during the study of initial screening medium and re-screening culture medium.More than 2000 strains of P-dissolving microbes were isolated from different soil samples by solid plate method .Results are shown as follows.P-dissolving microbes are most in vegetable garden soil because of its high content of organic matter.Quantity and kinds of P-dissolving microbes are different in soil from different area.Bacillus P17,Brevibacterium P10,Rhodotorula Y3 are isolated and identified.P-dissolving microbes could be isolated with the screening model by solid plate method,but P-dissolving effect is still needed to be determined combined with liquid screening model.%磷是植物体内有机化合物的主要成分,为生物固氮所必需,土壤中大部分磷以难溶态存在,不能被植物直接吸收利用,解磷微生物可以促进土壤难溶磷向有效磷的转化。基于目前耕地有效磷缺乏的现状,通过对解磷微生物初筛、复筛培养基的研究,探索了解磷微生物的筛选模型以及各地解磷微生物的多样性,对不同类型土壤中分离出的2000多株解磷微生物的溶磷效果进行了研究。结果表明:菜园土中有机质含量高,解磷微生物最多,从全国各地土样及种子表皮中分离出芽孢杆菌P17菌株、短杆菌P10菌株和红酵母Y3菌株等解磷微生物;不同地域、同种地区不同类型土壤中溶解难溶性无机磷和有机磷微生物的种类和数量存在显著差异;

  6. Cytosine deaminase as a negative selectable marker for the microalgal chloroplast: a strategy for the isolation of nuclear mutations that affect chloroplast gene expression.

    Science.gov (United States)

    Young, Rosanna E B; Purton, Saul

    2014-12-01

    Negative selectable markers are useful tools for forward-genetic screens aimed at identifying trans-acting factors that are required for expression of specific genes. Transgenic lines harbouring the marker fused to a gene element, such as a promoter, may be mutagenized to isolate loss-of-function mutants able to survive under selection. Such a strategy allows the molecular dissection of factors that are essential for expression of the gene. Expression of individual chloroplast genes in plants and algae typically requires one or more nuclear-encoded factors that act at the post-transcriptional level, often through interaction with the 5' UTR of the mRNA. To study such nuclear control further, we have developed the Escherichia coli cytosine deaminase gene codA as a conditional negative selectable marker for use in the model green alga Chlamydomonas reinhardtii. We show that a codon-optimized variant of codA with three amino acid substitutions confers sensitivity to 5-fluorocytosine (5-FC) when expressed in the chloroplast under the control of endogenous promoter/5' UTR elements from the photosynthetic genes psaA or petA. UV mutagenesis of the psaA transgenic line allowed recovery of 5-FC-resistant, photosynthetically deficient lines harbouring mutations in the nuclear gene for the factor TAA1 that is required for psaA translation. Similarly, the petA line was used to isolate mutants of the petA mRNA stability factor MCA1 and the translation factor TCA1. The codA marker may be used to identify critical residues in known nuclear factors and to aid the discovery of additional factors required for expression of chloroplast genes.

  7. Multi-species sequence comparison reveals dynamic evolution of the elastin gene that has involved purifying selection and lineage-specific insertions/deletions

    Directory of Open Access Journals (Sweden)

    Green Eric D

    2004-05-01

    Full Text Available Abstract Background The elastin gene (ELN is implicated as a factor in both supravalvular aortic stenosis (SVAS and Williams Beuren Syndrome (WBS, two diseases involving pronounced complications in mental or physical development. Although the complete spectrum of functional roles of the processed gene product remains to be established, these roles are inferred to be analogous in human and mouse. This view is supported by genomic sequence comparison, in which there are no large-scale differences in the ~1.8 Mb sequence block encompassing the common region deleted in WBS, with the exception of an overall reversed physical orientation between human and mouse. Results Conserved synteny around ELN does not translate to a high level of conservation in the gene itself. In fact, ELN orthologs in mammals show more sequence divergence than expected for a gene with a critical role in development. The pattern of divergence is non-conventional due to an unusually high ratio of gaps to substitutions. Specifically, multi-sequence alignments of eight mammalian sequences reveal numerous non-aligning regions caused by species-specific insertions and deletions, in spite of the fact that the vast majority of aligning sites appear to be conserved and undergoing purifying selection. Conclusions The pattern of lineage-specific, in-frame insertions/deletions in the coding exons of ELN orthologous genes is unusual and has led to unique features of the gene in each lineage. These differences may indicate that the gene has a slightly different functional mechanism in mammalian lineages, or that the corresponding regions are functionally inert. Identified regions that undergo purifying selection reflect a functional importance associated with evolutionary pressure to retain those features.

  8. Selection affects genes involved in replication during long-term evolution in experimental populations of the bacteriophage φX174.

    Directory of Open Access Journals (Sweden)

    Celeste J Brown

    Full Text Available Observing organisms that evolve in response to strong selection over very short time scales allows the determination of the molecular mechanisms underlying adaptation. Although dissecting these molecular mechanisms is expensive and time-consuming, general patterns can be detected from repeated experiments, illuminating the biological processes involved in evolutionary adaptation. The bacteriophage φX174 was grown for 50 days in replicate chemostats under two culture conditions: Escherichia coli C as host growing at 37°C and Salmonella typhimurium as host growing at 43.5°C. After 50 days, greater than 20 substitutions per chemostat had risen to detectable levels. Of the 97 substitutions, four occurred in all four chemostats, five arose in both culture conditions, eight arose in only the high temperature S. typhimurium chemostats, and seven arose only in the E. coli chemostats. The remaining substitutions were detected only in a single chemostat, however, almost half of these have been seen in other similar experiments. Our findings support previous studies that host recognition and capsid stability are two biological processes that are modified during adaptation to novel hosts and high temperature. Based upon the substitutions shared across both environments, it is apparent that genome replication and packaging are also affected during adaptation to the chemostat environment, rather than to temperature or host per se. This environment is characterized by a large number of phage and very few hosts, leading to competition among phage within the host. We conclude from these results that adaptation to a high density environment selects for changes in genome replication at both protein and DNA sequence levels.

  9. Isolation and cultivation of microalgae select for low growth rate and tolerance to high pH

    DEFF Research Database (Denmark)

    Berge, Terje; Daugbjerg, Niels; Hansen, Per Juel

    2012-01-01

    Harmful microalgal blooms or red tides are often associated with high levels of pH. Similarly, species and strains of microalgae cultivated in the laboratory with enriched media experience recurrent events of high pH between dilutions with fresh medium. To study the potential for laboratory...... of upper pH tolerance limits were higher in the younger (20 years). These results suggest selection of strains best adapted to tolerate or postpone/avoid events of high pH in the laboratory. Our data have implications for experimental studies of pH response and reaction norms in general of microalgae...

  10. Transcriptome-wide N⁶-methyladenosine profiling of rice callus and leaf reveals the presence of tissue-specific competitors involved in selective mRNA modification.

    Science.gov (United States)

    Li, Yuli; Wang, Xiliang; Li, Cuiping; Hu, Songnian; Yu, Jun; Song, Shuhui

    2014-01-01

    N(6)-methyladenosine (m(6)A) is the most prevalent internal modification present in mRNAs of all higher eukaryotes. With the development of MeRIP-seq technique, in-depth identification of mRNAs with m(6)A modification becomes feasible. Here we present a transcriptome-wide m(6)A modification profiling effort for rice transcriptomes of differentiated callus and leaf, which yields 8,138 and 14,253 m(6)A-modified genes, respectively. The m(6)A peak (m(6)A-modified nucleotide position on mRNAs) distribution exhibits preference toward both translation termination and initiation sites. The m(6)A peak enrichment is negatively correlated with gene expression and weakly positively correlated with certain gene features, such as exon length and number. By comparing m(6)A-modified genes between the 2 samples, we define 1,792 and 6,508 tissue-specific m(6)A-modified genes (TSMGs) in callus and leaf, respectively. Among which, 626 and 5,509 TSMGs are actively expressed in both tissues but are selectively m(6)A-modified (SMGs) only in one of the 2 tissues. Further analyses reveal characteristics of SMGs: (1) Most SMGs are differentially expressed between callus and leaf. (2) Two conserved RNA-binding motifs, predicted to be recognized by PUM and RNP4F, are significantly over-represented in SMGs. (3) GO enrichment analysis shows that SMGs in callus mainly participate in transcription regulator/factor activity whereas SMGs in leaf are mainly involved in plastid and thylakoid. Our results suggest the presence of tissue-specific competitors involved in SMGs. These findings provide a resource for plant RNA epitranscriptomic studies and further enlarge our knowledge on the function of RNA m(6)A modification.

  11. Characterization of Neisseria cinerea, a nonpathogenic species isolated on Martin-Lewis medium selective for pathogenic Neisseria spp.

    Science.gov (United States)

    Knapp, J S; Totten, P A; Mulks, M H; Minshew, B H

    1984-01-01

    An asaccharolytic, gram-negative, oxidase-positive diplococcus was isolated on Martin-Lewis medium from the cervix of a patient attending an arthritis clinic at Seattle Public Health Hospital, Seattle, Wash. This strain, NRL 32165, did not produce detectable acid from glucose, maltose, sucrose, fructose, mannitol, or lactose in either cystine Trypticase agar (BBL Microbiology Systems, Cockeysville, Md.) or modified oxidation-fermentation medium and was identified presumptively as a glucose-negative Neisseria gonorrhoeae strain, but was identified later as Neisseria cinerea on the basis of its biochemical reactions. Nitrate was not reduced, nitrite (0.001%, wt/vol) was reduced, and polysaccharide was not produced from sucrose. Proline, arginine, and cystine-cysteine were required for growth on defined medium. Strain NRL 32165 did not react with antigonococcal protein I monoclonal antibodies and did not produce immunoglobulin A protease. In DNA:DNA homology studies with N. gonorrhoeae NRL 8038 (F62) and N. cinerea type strain NRL 30003, strain NRL 32165 showed 95% homology relative to N. cinerea and 44% homology relative to N. gonorrhoeae. Thus, the identity of strain NRL 32165 was confirmed as N. cinerea (von Lingelsheim 1906) Murray 1939. Of all Neisseria spp., N. cinerea is most likely to be misidentified as a glucose-negative N. gonorrhoeae strain.

  12. Phytochemical and antibacterial activity of the extracts of Fagara zanthoxyloides on selected cariogenic and enteric bacterial isolates

    Directory of Open Access Journals (Sweden)

    Martins Ajibade Adefisoye

    2012-02-01

    Methods: Crude extracts were obtained by cold extraction method of the powdered stem in methanol-water mixture (MW in ratio 3:2 and phosphate buffer saline (PBS. Filtrates obtained were concentrated in a rotary evaporator and lyophilized. Antimicrobial activity of the extracts, at a concentration of 25 mg/ml was tested against four bacterial isolates using agar well diffusion method. Phytochemical analysis of the plant extract for the presence of tannins, saponins, alkaloids and flavanoids was based on chemical examination. Results and discussions: Extracts from methanol-water mixture showed some antimicrobial activity against Lactobacillus brevis (NCIMB 4617, L. plantarum (NCDO 1752 and Escherichia coli and Proteus vulgaris. The minimum inhibitory concentration (MIC of extracts ranged between 1.57 and 12.5 mg/ml except in E. coli with value greater than 12.5.mg/ml. The phytochemical screening indicated that the extract tested positive for tannin, saponin, flavonoids and alkaloids. Results showed that F. zanthoxyloides would be valuable in the treatment of microbial diseases particularly those of the oral cavity. The considerable antibacterial activities exhibited by the extract of the plant thus justify the use as a teeth cleansing agent in the local setting. The problem of antibiotics resistance to synthetic drugs is also expected to be mitigated with the use of natural drugs. [J Intercult Ethnopharmacol 2012; 1(1.000: 1-6

  13. A coordination polymer based magnetic adsorbent material for hemoglobin isolation from human whole blood, highly selective and recoverable

    Science.gov (United States)

    Zhang, Xiaoxing; Tan, Jipeng; Xu, Xinxin; Shi, Fanian; Li, Guanglu; Yang, Yiqiao

    2017-09-01

    A composite material has been obtained successfully through the loading of nanoscale coordination polymer on magnetic Fe3O4@SiO2 core-shell particle. In this composite material, coordination polymer nanoparticles distribute uniformly on Fe3O4@SiO2 and these two components are ;tied; together firmly with chemical bonds. Adsorption experiments suggest this composite material exhibits very excellent selectivity to hemoglobin. But under the same condition, its adsorption to bovine serum albumin can almost be ignored. This selectivity can be attributed to the existence of hydrophobic interactions between coordination polymer nanoparticle and hemoglobin. For composite material, the hemoglobin adsorption process follows Langmuir model perfectly with high speed. The adsorbed hemoglobin can be eluted easily by sodium dodecyl sulfate stripping reagent with structure and biological activity of hemoglobin keeps well. The composite material was also employed to separate hemoglobin from human whole blood, which receives a very satisfactory result. Furthermore, magnetic measurement reveals ferromagnetic character of this composite material with magnetization saturation 3.56 emu g-1 and this guarantees its excellent magnetic separation performance from the treated solution.

  14. Selective growth inhibition of human breast cancer cells by graviola fruit extract in vitro and in vivo involving downregulation of EGFR expression.

    Science.gov (United States)

    Dai, Yumin; Hogan, Shelly; Schmelz, Eva M; Ju, Young H; Canning, Corene; Zhou, Kequan

    2011-01-01

    The epidermal growth factor receptor (EGFR) is an oncogene frequently overexpressed in breast cancer (BC), and its overexpression has been associated with poor prognosis and drug resistance. EGFR is therefore a rational target for BC therapy development. This study demonstrated that a graviola fruit extract (GFE) significantly downregulated EGFR gene expression and inhibited the growth of BC cells and xenografts. GFE selectively inhibited the growth of EGFR-overexpressing human BC (MDA-MB-468) cells (IC(50) = 4.8 μg/ml) but had no effect on nontumorigenic human breast epithelial cells (MCF-10A). GFE significantly downregulated EGFR mRNA expression, arrested cell cycle in the G0/G1 phase, and induced apoptosis in MDA-MB-468 cells. In the mouse xenograft model, a 5-wk dietary treatment of GFE (200 mg/kg diet) significantly reduced the protein expression of EGFR, p-EGFR, and p-ERK in MDA-MB-468 tumors by 56%, 54%, and 32.5%, respectively. Overall, dietary GFE inhibited tumor growth, as measured by wet weight, by 32% (P < 0.01). These data showed that dietary GFE induced significant growth inhibition of MDA-MB-468 cells in vitro and in vivo through a mechanism involving the EGFR/ERK signaling pathway, suggesting that GFE may have a protective effect for women against EGFR-overexpressing BC.

  15. Enteroaggregative Escherichia coli isolated from Chinese diarrhea patients with high-pathogenicity island of Yersinia is involved in synthesis of siderophore yersiniabactin

    Institute of Scientific and Technical Information of China (English)

    Jing Hu; Biao Kan; Zhi-Hua Liu; Shou-Yi Yu

    2005-01-01

    AIM: To investigate the distribution of 12 high-pathogenicity island (HPI) genes and the relation between HPI genes and expression of yersiniabactin (Ybt) in enteroaggregative E.coli(EAggEC) isolated from Chinese diarrhea patients.METHODS: The distribution of 12 HPI genes was investigated by PCR and DNA hybridization in two prototype strains of EAggEC, EAggEC 17-2, EAggEC O42, and 6 clinical EAggEC isolates from China. The production of siderophore Ybt in HPI-positive strains was detected by reporter gene bioassay to determine the relation between HPI genes and expression of Ybt. Flow cytometry was used to detect fluorescent signal of the reporter strain that could designate production of Ybt.RESULTS: Seven strains were HPI-positive and one strain was HPI-negative. Six of the seven HPI-positive strains were inserted into asnT-tRNA site. Moreover, seven EAggEC HPI-positive strains revealed enhanced fluorescence signal but the EAggEC HPI-negative strain did not. However, there was a difference in Ybt expression condition and level among these seven EAggEC HPI-positive strains. Although UFT073 strain, the prototype strain of uropathogenic E.coli(UPEC), carried the complete HPI core part, we did not detect the expression of Ybt in it.CONCLUSION: EAggEC HPI-positive strains can express the Ybt system, but the presence of HPI core part does not mean the functional expression of Ybt.

  16. LA-ICP-MS analysis of isolated phosphatic grains indicates selective rare earth element enrichment during reworking and transport processes

    Science.gov (United States)

    Auer, Gerald; Reuter, Markus; Hauzenberger, Christoph A.; Piller, Werner E.

    2016-04-01

    Rare earth elements (REE) are a commonly used proxy to reconstruct water chemistry and oxygen saturation during the formation history of authigenic and biogenic phosphates in marine environments. In the modern ocean REE exhibit a distinct pattern with enrichment of heavy REE and strong depletion in Cerium. Studies of ancient phosphates and carbonates, however, showed that this 'modern' pattern is only rarely present in the geological past. Consequently, the wide range of REE enrichment patterns found in ancient marine phosphates lead to the proposition that water chemistry had to have been radically different in the earth's past. A wealth of studies has already shown that both early and late diagenesis can strongly affect REE signatures in phosphates and severely alter primary marine signals. However, no previous research was conducted on how alteration processes occurring prior to final deposition affect marine phosphates. Herein we present a dataset of multiple LA-ICP-MS measurements of REE signatures in isolated phosphate and carbonate grains deposited in a carbonate ramp setting in the central Mediterranean Sea during the middle Miocene "Monterey event". The phosphates are represented by authigenic, biogenic and detrital grains emplaced in bioclastic grain- to packstones dominated by bryozoan and echinoderm fragments, as well as abundant benthic and planktic foraminifers. The results of 39 grain specific LA-ICP-MS measurements in three discrete rock samples reveals four markedly different REE patterns (normalized to the Post Archean Australian Shale standard) in terms of total enrichment and pattern shape. Analyses of REE diagenesis proxies show that diagenetic alteration affected the samples only to a minor degree. Considering grain shape and REE patterns together indicate that authigenic, detrital and biogenic phosphates have distinct REE patterns irrespective of the sample. Our results show that the observed REE patterns in phosphates only broadly reflect

  17. 烟草疫霉分离及生长培养基的选择%Selection of Isolating and Culturing Medium for Phytophthora nicotianae

    Institute of Scientific and Technical Information of China (English)

    苏凯; 桑维钧; 张新强; 王慧

    2013-01-01

    Different base medium adding different combination of antibiotics were applied for isolation and culture of Phytophthora nicotianae.The results showed that selective medium with Vs base medium (100mL/L V8juice+0.02 g/L CaCO3+2 g/Lagar) adding 10 mg/mL ampicillin +5 mg/mL nysfungin+5 mg/mL carbendazim had the best isolation effect as the successful isolation rate was 100%,and the hypha grew well on this medium.After separation,the hypha could be cultured in Vs base medium as the hypha was white,tight and strong with big bacteria clone and strong growth potential.%采用不同的基本培养基添加抗生素组合对烟草疫霉(Phytophthora nicotianae)进行分离培养.结果表明,以V8培养基(100 mL/L Vs汁+0.02 g/L CaCO3+2 g/L洋菜)为基本培养基附加10 mg/mL氨苄青霉素+5 mg/mL制霉素+5 mg/mL多菌灵的选择培养基分离烟草疫霉的效果较好,分离成功率达100%,菌丝的生长状态也较好.菌丝分离后在V8基本培养基中培养,菌丝紧密、浓白、粗壮,菌落大、长势旺.

  18. Clinical features of AIDS patients with Hodgkin’s lymphoma with isolated bone marrow involvement:report of 12 cases at a single institution

    Institute of Scientific and Technical Information of China (English)

    Marcelo Corti; Maria Villafae; Gonzalo Minue; Ana Campitelli; Marina Narbaitz; Leonardo Gilardi

    2015-01-01

    Objective:To study the main clinical and histopathological features of 12 patients with Hodgkin’s lymphoma (HL) diagnosed primarily from bone marrow (BM) involvement. Methods:We included 12 acquired immunodeficiency syndrome (AIDS) patients with HL assisted in the F. J. Muñiz Infectious Diseases Hospital since January 2002 to December 2013. hTe diagnosis of HL with primary BM involvement in patients was conifrmed by clinical, histopathological, and immunohistochemical ifndings. Results:All patients presented“B”symptoms and pancytopenia. All of them had stage IV neoplasm disease because of BM inifltration. hTe median of CD4+T-cell counts was 114 cells/μL, and mixed cellularity (MC) was the most frequent histopathological subtype of 92%cases. Conclusion:When other causes are excluded, BM biopsy should be performed in AIDS patients with“B”symptoms and pancytopenia to evaluate BM inifltration by atypical lymphocytes.

  19. Selection, isolation and growth kinetic study of a bacterial consortium obtained from the Potengi mangrove in the presence of crude oil

    Energy Technology Data Exchange (ETDEWEB)

    Costa, C.C.; Vaz, M.R.F.; Santos, E.S.; Macedo, G.R. [Universidade Federal do Rio Grande do Norte (UFRN), Natal, RN (Brazil). Dept. de Engenharia Quimica], E-mail: natcintia@gmail.com; Costa, J.G. da [Universidade Federal do Amazonas (UFAM), Coari, AM (Brazil). Inst. de Saude e Biotecnologia

    2011-10-15

    The selection, isolation and kinetic study of a bacterial consortium obtained from a sample of soil from the Potengi mangrove, located in the city of Natal, Rio Grande do Norte, Brazil, has been carried out using the enrichment culture technique to observe aspects such as the evaluation of main growth parameters. The kinetic study used a rotary incubator shaker at 150rpm, under 30 deg C. The bacterial consortium isolated from the estuary of the Potengi River showed a good acclimation in minimum mineral medium with 1% (v/v) of oil. The cell concentration reached 2.55 g/L at 16h of cultivation and surface tension dropped. The maximum productivity in cells obtained was of 0.3 g/L.h, the specific velocity of growth was of 0.075h{sup -1}, with a generation time (tg) of 9.24h. This study seeks to demonstrate that the consortium can be used as inoculants in biological treatments, capable of reducing the waste's degradation time. (author)

  20. Two new proanthocyanidin trimers isolated from Cistus incanus L. demonstrate potent anti-inflammatory activity and selectivity to cyclooxygenase isoenzymes inhibition.

    Science.gov (United States)

    Mansoor, K A; Matalka, K Z; Qa'dan, F S; Awad, R; Schmidt, M

    2016-09-01

    Two new proanthocyanidin trimers have been isolated from Cistus incanus herb; gallocatechin-(4α→6)-gallocatechin-(4α→8)-gallocatechin (compound 1) and epigallocatechin-3-O-gallate-(4ß→8)-epigallocatechin-3-O-gallate-(4ß→8)-gallocatechin (compound 2). The structures were determined on the basis of 1D- and 2D-NMR (HSQC, HMBC) of their peracetylated derivatives, MALDI-TOF-MS and by acid-catalysed degradation with phloroglucinol. A more abundant proanthocyanidin oligomer was also isolated, purified and its chemical constitution studied by (13)C-NMR and phloroglucinol degradation. The mean molecular weight of the polymer was estimated to be about 7 to 8 flavan-3-ol-units with a ratio of procyanidin : prodelphinidin units at 1:5, some of which are derivatised by gallic acid. Water extract and higher oligomeric proanthocyanidin fractions of C. incanus significantly inhibited TPA-induced oedema when applied topically at doses of 0.5 and 1 mg/ear in mice. Furthermore, the extracts and the pure compounds inhibited COX-1 and COX-2 activities. In addition, compound 2 exhibited an IC50 of 4.5 μM against COX-2 indicating its high selectivity towards COX-2.

  1. Isolated Granulocytic Sarcoma of the Breast after Allogeneic Stem Cell Transplantation: A Rare Involvement Also Detected by 18FDG-PET/CT

    Directory of Open Access Journals (Sweden)

    Eren Gündüz

    2014-03-01

    Full Text Available Granulocytic sarcoma is a tumor consisting of myeloid blasts with or without maturation that occurs at an anatomical site other than bone marrow. Most frequently affected sites are skin, lymph nodes, gastrointestinal tract, bone, soft tissue and testes. AML may manifest as granulocytic sarcoma at diagnosis or relapse. Although it has been considered to be rare relapse as granulocytic sarcoma after stem cell transplantation is being increasingly reported. However it is rare without bone marrow involvement and in AML M6 subtype. Breast is also a rare involvement. We report a 30-year-old woman with AML M6 relapsed 16 months after allogeneic stem cell transplantation as a granulocytic sarcoma in right breast without bone marrow involvement. She was treated with systemic chemotherapy but died of sepsis. 18FDG-PET/CT images were also obtained and detected lesions other than detected by breast ultrasound. The incidence of granulocytic sarcoma may increase if suspected or new diagnostic modalities are performed.

  2. Simultaneous Counter-Ion Co-Deposition a Technique Enabling Matrix Isolation Spectroscopy Studies Using Low-Energy Beams of Mass-Selected Ions

    Science.gov (United States)

    Ludwig, Ryan M.; Moore, David T.

    2014-06-01

    Matrix isolation spectroscopy was first developed in Pimentel's group during the 1950's to facilitate spectroscopic studies of transient species. Cryogenic matrices of condensed rare gases provide an inert chemical environment with facile energy dissipation and are transparent at all wavelengths longer than vacuum UV, making them ideal for studying labile and reactive species such as radicals, weakly bound complexes, and ions. Since frozen rare gases are poor electrolytes, studies of ions require near-equal populations of anions and cations in order to stabilize the number densities required for spectroscopic experiments. Many techniques for generation of ions for using in matrix isolation studies satisfy this criterion intrinsically, however when ion beams generated in external sources are deposited, the counter-ions typically arise via secondary processes that are at best loosely controlled. It has long been recognized that it would be desirable to stabilize deposition of mass-selected ions generated in an external source using simultaneous co-deposition of a beam of counter-ions, however previous attempts to achieve this have been reported as unsuccessful. The Moore group at Lehigh has demonstrated successful experiments of this type, using mass-selected anions generated from a metal cluster source, co-deposited with a balanced current of cations generated in a separate electron ionization source. This talk will focus on the details of the technique, and present some results from proof-of-concept studies on anionic copper carbonyl complexes formed in argon matrices following co-deposition of Cu- with Ar+ or Kr+. Funding support from NSF CAREER Award CHE-0955637 is gratefully acknowledged. Whittle et al., J. Chem. Phys. 22, p.1943 (1954); Becker et al., J. Chem. Phys. 25, p.224 (1956). Godbout et al., J. Chem. Phys. 96, p.2892 (1996). Sabo et al., Appl. Spectrosc. 45, p. 535 (1991).

  3. Isolation of a selected microbial consortium capable of degrading methyl parathion and p-nitrophenol from a contaminated soil site.

    Science.gov (United States)

    Pino, Nancy J; Dominguez, Maria C; Penuela, Gustavo A

    2011-01-01

    A bacterial consortium with the ability to degrade methyl parathion and p-nitrophenol, using these compounds as the only carbon source, was obtained by selective enrichment in a medium with methyl parathion. Samples were taken from Moravia, Medellin; an area that is highly contaminated, owing to the fact that it was used as a garbage dump from 1974 to 1982. Acinetobacter sp, Pseudomonas putida, Bacillus sp, Pseudomonas aeruginosa Citrobacter freundii, Stenotrophomonas sp, Flavobacterium sp, Proteus vulgaris, Pseudomonas sp, Acinetobacter sp, Klebsiella sp and Proteus sp were the microorganisms identified within the consortium. In culture, the consortium was able to degrade 150 mg L⁻¹ of methyl-parathion and p-nitrophenol in 120 h, but after adding glucose or peptone to the culture, the time of degradation decreased to 24 h. In soil, the consortium was also able to degrade 150 mg L⁻¹ of methyl parathion in 120 h at different depths and also managed to decrease the toxicity.

  4. Selective Effect of 2′,6′-Dihydroxy-4′-Methoxychalcone Isolated from Piper aduncum on Leishmania amazonensis

    Science.gov (United States)

    Torres-Santos, Eduardo Caio; Moreira, Davyson Lima; Kaplan, Maria Auxiliadora C.; Meirelles, Maria Nazareth; Rossi-Bergmann, Bartira

    1999-01-01

    2′,6′-Dihydroxy-4′-methoxychalcone (DMC) was purified from the dichloromethane extract of Piper aduncum inflorescences. DMC showed significant activity in vitro against promastigotes and intracellular amastigotes of Leishmania amazonensis, with 50% effective doses of 0.5 and 24 μg/ml, respectively. Its inhibitory effect on amastigotes is apparently a direct effect on the parasites and is not due to activation of the nitrogen oxidative metabolism of macrophages, since the production of nitric oxide by both unstimulated and recombinant gamma interferon-stimulated macrophages was decreased rather than increased with DMC. The phagocytic activity of macrophages was functioning normally even with DMC concentrations as high as 80 μg/ml, as seen by electron microscopy and by the uptake of fluorescein isothiocyanate-labeled beads. Ultrastructural studies also showed that in the presence of DMC the mitochondria of promastigotes were enlarged and disorganized. Despite destruction of intracellular amastigotes, no disarrangement of macrophage organelles were observed, even at 80 μg of DMC/ml. These observations suggest that DMC is selectively toxic to the parasites. Its simple structure may well enable it to serve as a new lead compound for the synthesis of novel antileishmanial drugs. PMID:10223942

  5. Environmental concentrations of the selective serotonin reuptake inhibitor fluoxetine impact specific behaviors involved in reproduction, feeding and predator avoidance in the fish Pimephales promelas (fathead minnow).

    Science.gov (United States)

    Weinberger, Joel; Klaper, Rebecca

    2014-06-01

    Pharmaceuticals and personal care products (PPCPs) have been found in surface waters worldwide, but little is understood of their effects on the wildlife that inhabit these waters. Fluoxetine (Prozac; Eli Lilly), a highly prescribed selective serotonin reuptake inhibitor (SSRI), is a commonly found PPCP in surface water. The purpose of this project was to determine if environmentally relevant concentrations of fluoxetine impact behavior that is important for population survival in native fish species, including reproduction, feeding and predator avoidance. Chronic 4-week exposures were conducted with doses ranging from 100 ng/L to 100 μg/L to cover a range of environmentally relevant concentrations up to higher concentrations comparable to other published studies with the same drug that have documented various physiological impacts. Pimephales promelas (fathead minnow), a species native to North America, was used as it conducts a range of specific mating behaviors and therefore serves as an excellent model of specific impacts on brain function. Fluoxetine concentrations as low as 1 μg/L, a concentration that has been found in many freshwater environments, were found to significantly impact mating behavior, specifically nest building and defending in male fish. Males were also found to display aggression, isolation, and repetitive behaviors at higher concentrations. Female mating behavior was largely unaffected. In addition, predator avoidance behaviors in males and females were also impacted at 1 μg/L. Feeding was impacted at 10 μg/L and in the highest exposure (100 μg/L), egg production was limited by deaths of females due to significant male aggressive behaviors in the first two weeks of exposure. Specific behavioral changes occurred at each concentration (most noticeably 1 μg/L and 100 μg/L) indicating a dose dependent effect that triggered different responses at lower exposures versus higher exposures or differential impacts of dose depending on brain

  6. Monoclonal antibody-based, selective isolation of DNA fragments containing an alkylated base to be quantified in defined gene sequences.

    Science.gov (United States)

    Hochleitner, K; Thomale, J; Nikitin AYu; Rajewsky, M F

    1991-08-25

    We have established a sensitive, monoclonal antibody (Mab)-based procedure permitting the selective enrichment of sequences containing the miscoding alkylation product O6-ethylguanine (O6-EtGua) from mammalian DNA. H5 rat hepatoma cells were reacted with the N-nitroso carcinogen N-ethyl-N-nitrosourea in vitro, to give overall levels of greater than or equal to 25 O6-EtGua residues per diploid genome (corresponding to O6-EtGua/guanine molar ratios of greater than or equal to 10(-8). For analysis, enzymatically restricted DNA from these cells is incubated with an antibody specific for O6-ethyl-2'-deoxyguanosine, the resulting Mab-DNA complexes are separated from (O6-EtGua)-free fragments by filtration through a nitrocellulose (NC) membrane, and the DNA is recovered from the filter-bound complexes quantitatively. The efficiency of Mab binding to DNA fragments containing O6-EtGua is constant over a range of O6-EtGua/guanine molar ratios between 10(-5) and 10(-8). (O6-EtGua)-containing restriction fragments encompassing known gene sequences (e.g., the immunoglobulin E heavy chain gene of H5 rat hepatoma cells used as a model in this study) are subsequently amplified by PCR and quantified by slot-blot hybridisation. The content and distribution of a specific carcinogen-DNA adduct in defined sequences of genomic DNA can thus be analyzed as well as the kinetics of intragenomic (toposelective) repair of any DNA lesion for which a suitable Mab is available.

  7. Development of a new transformant selection system for Penicillium chrysogenum: isolation and characterization of the P. chrysogenum acetyl-coenzyme A synthetase gene (facA) and its use as a homologous selection marker.

    Science.gov (United States)

    Gouka, R J; van Hartingsveldt, W; Bovenberg, R A; van Zeijl, C M; van den Hondel, C A; van Gorcom, R F

    1993-01-01

    A new transformation system for the filamentous fungus Penicillium chrysogenum is described, based on the use of the homologous acetyl-coenzyme A synthetase (facA) gene as a selection marker. Acetate-non-utilizing (Fac-) strains of P. chrysogenum were obtained by positive selection for spontaneous resistance to fluoroacetate. Among these fac mutants putative facA strains were selected for a loss of acetyl-coenzyme A (CoA) synthetase activity. The facA gene, coding for the enzyme acetyl-CoA synthetase, was isolated from a P. chrysogenum genomic library using synthetic oligonucleotides derived from conserved regions from the corresponding genes of Aspergillus nidulans and Neurospora crassa. Vector pPC2-3, comprising a genomic 6.5 kb PstI fragment, was able to complement P. chrysogenum facA strains with frequencies up to 27 transformants.micrograms-1 DNA. Direct selection of transformants was accomplished using acetate and low amounts (0.001%) of glucose as carbon sources. About 50% of the transformants arose by integration of pPC2-3 DNA at the homologous facA locus and 50% by integration elsewhere in the genome. Determination of the nucleotide sequence of part of the cloned fragment showed the presence of an open reading frame of 2007 nucleotides, interrupted by five putative introns. Comparison of the nucleotide and the amino acid sequence of the facA gene of P. chrysogenum with the facA gene of A. nidulans reveals similarities of 80% and 89%, respectively. The putative introns present in the P. chrysogenum facA gene appear at identical positions as those in the A. nidulans facA gene, but show no significant sequence similarity.

  8. Gonosomal mosaicism in myotonic dystrophy patients: Involvement of mitotic events in (CTG)[sub n] repeat variation and selection against extreme expansion in sperm

    Energy Technology Data Exchange (ETDEWEB)

    Jansen, G.; Coerwinkel, M.; Wieringa, B.; Nillesen, W.; Smeets, H.; Brunner, H.; Wieringa, B. (Univ. of Nijmegen (Netherlands)); Willems, P.; Vits, L. (Univ. of Antwerp (Belgium)); Hoeweler, C. (Univ. of Maastricht (Netherlands))

    1994-04-01

    Myotonic dystrophy (DM) is caused by abnormal expansion of a polymorphic (CTG)[sub n] repeat, located in the DM protein kinase gene. The authors determined the (CTG)[sub n] repeat lengths in a broad range of tissue DNAs from patients with mild, classical, or congenital manifestation of DM. Differences in the repeat length were seen in somatic tissues from single DM individuals and twins. Repeats appeared to expand to a similar extent in tissues originating from the same embryonal origin. In most male patients carrying intermediate- or small-sized expansions in blood, the repeat lengths covered a markedly wider range in sperm. In contrast, male patients with large allele expansions in blood (>700 CTGs) had similar or smaller repeats in sperm, when detectable. Sperm alleles with >1,000 CTGs were not seen. The authors conclude that DM patients can be considered gonosomal mosaics, i.e., combined somatic and germ-line tissue mosaics. Most remarkably, they observed multiple cases where the length distributions of intermediate- or small-sized alleles in fathers' sperm were significantly different from that in their offspring's blood. The combined findings indicate that intergenerational length changes in the unstable CTG repeat are most likely to occur during early embryonic mitotic divisions in both somatic and germ-line tissue formation. Both the initial CTG length, the overall number of cell divisions involved in tissue formation, and perhaps a specific selection process in spermatogenesis may influence the dynamics of this process. A model explaining mitotic instability and sex-dependent segregation phenomena in DM manifestation is discussed. 59 refs., 5 figs.

  9. Either the dorsal hippocampus or the dorsolateral striatum is selectively involved in consolidation of forced swim-induced immobility depending on genetic background.

    Science.gov (United States)

    Colelli, V; Campus, P; Conversi, D; Orsini, C; Cabib, S

    2014-05-01

    Healthy subjects differ in the memory system they engage to learn dual-solution tasks. Both genotype and stress experience could contribute to this phenotypic variability. The present experiments tested whether the hippocampus and the dorsal striatum, the core nodes of two different memory systems, are differently involved in 24 h retention of a stress-associated memory in two genetically unrelated inbred strains of mice. Mice from both the C57BL/6J and the DBA/2J inbred strains showed progressive increase of immobility during 10 min exposure to forced swim (FS) and retrieved the acquired levels of immobility when tested 24h later. The pattern of c-fos immunostaining promoted by FS revealed activation of a large number of brain areas in both strains, including CA1 and CA3 fields of the hippocampus. However, only DBA/2J mice showed activation of the dorsolateral striatum (DLS). In addition, FS induced a positive correlation between c-fos expression in the amygdala and CA1 and CA3 in C57BL/6J mice whereas it induced a positive correlation between c-fos expression in the amygdala and DLS in DBA/2J mice. Finally, temporary post-training inactivation of the dorsal hippocampus, by local infusion of lidocaine, prevented 24h retention of immobility in C57BL/6J mice only, whereas inactivation of the DLS prevented retention in DBA/2J mice only. These findings support the view that genetic factors can determine whether the dorsal hippocampus or the DLS are selectively engaged to consolidate stress-related memory.

  10. Investigating the influence of selected texture-improved pretreatment techniques on storage stability of wholegrain brown rice: Involvement of processing-induced mineral changes with lipid degradation.

    Science.gov (United States)

    Xia, Qiang; Wang, Liping; Yu, Wenjuan; Li, Yunfei

    2017-09-01

    This work aimed at investigating the effects of emerging texture-improved processing techniques including high hydrostatic pressure (HHP; 150-450MPa/10min), high-intensity ultrasonication (HIU; 17.83W·cm(-2)/30min) and germination (37°C/36h) pretreatments on lipid hydrolysis and oxidation development of wholegrain brown rice (WBR) during storage, in an attempt to ascertain a possible link between lipid degradation and the underlying mechanisms. The results showed that HHP and HIU treatments enhanced lipid hydrolysis and oxidation as indicated by the formation of free fatty acids (FFA) and thiobarbituric acid reactive substances (TBARS) respectively, whereas an opposite pattern was observed for germination. Storage process rather than after immediate treatments observed an increase in lipid oxidation of HHP and HIU-processed samples, which was related to processing-induced liberation of minerals. Quantitative and qualitative characterization via inductively coupled plasma-optical emission spectrometry (ICP-OES) and micro X-ray fluorescence (μ-XRF) analysis confirmed the shifts of mineral distribution in WBR grains in response to different pretreatments. The WBR-derived lipase was activated by Ca(2+), and μ-XRF mapping indicated calcium enrichment in pericarp/aleurone layer and its mobilization to embryo during germination process where magnesium and manganese were significantly reduced. Multivariate analysis revealed a close relationship between increased lipid degradation and minerals including magnesium and manganese. This is the first time for reporting the effects of selected texture-improved techniques on lipid stability of WBR grains across storage process as well as validating the involvement of processing-induced mineral release in lipid degradation. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Increased expressions of genes and proteins involved in mitochondrial oxidation and antioxidant pathway in adipose tissue of pigs selected for a low residual feed intake.

    Science.gov (United States)

    Louveau, I; Vincent, A; Tacher, S; Gilbert, H; Gondret, F

    2016-12-01

    Adipose tissue is a primary sensor for nutrient availability and regulates many functions including feed intake and energy homeostasis. This study was undertaken to determine the molecular responses of adipose tissue to differences in feed intake and feed efficiency. Subcutaneous adipose tissue was collected from two lines of pigs divergently selected for residual feed intake (RFI), a measure of feed efficiency defined as the difference between actual and expected feed intake, and from a subset of high-RFI pigs that were feed-restricted at the level of the voluntary feed intake of low-RFI pigs during the growing-finishing period. Transcriptomics analyses indicated that the number of genes that were differentially expressed ( feed-restricted high-RFI ( = 8) pigs. They included in silico pathway analyses of the differentially expressed (DE) genes ( feed restriction. This indicates that the responses of adipose tissue to RFI difference shared only some common mechanisms with feed intake modulation, notably the regulation of cell cycle (including ) and transferase activity pathway. Two carboxylesterase genes (, ) involved in lipolysis, were among the most overexpressed genes in the low-RFI pigs; they were also affected by feed restriction within the high-RFI line. About 60% of the molecular changes between low- and high-RFI pigs were specific to genetic divergence in feed efficiency, independently of feed intake. Different genes and proteins known to be associated with mitochondrial oxidative metabolism were overexpressed in adipose tissue of low-RFI pigs compared with high-RFI pigs; other proteins participating in the generation of energy were also affected by feed restriction within the high-RFI line. Finally, mitochondrial antioxidant genes were upregulated in low-RFI pigs vs. high-RFI pigs. Altogether, increased oxidative and antioxidant processes in adipose tissue might be associated with improved feed efficiency.

  12. Fungal strains isolated from cork stoppers and the formation of 2,4,6-trichloroanisole involved in the cork taint of wine.

    Science.gov (United States)

    Prak, Sina; Gunata, Ziya; Guiraud, Joseph-Pierre; Schorr-Galindo, Sabine

    2007-05-01

    Cork taint is mainly due to 2,4,6-trichloroanisole (TCA) produced through the activity of undesirable fungal strains. We observed that CFU mould number in TCA-containing stoppers was not quantitatively different to that of the stoppers not containing TCA (ca. 10(5)CFU/g). In contrast more fungi diversity was observed in TCA-containing stoppers. Penicillium spp (Penicillium chrysogenum, Penicillium glabrum), Aspergillus spp (Aspergillus niger and Aspergillus oryzae), Chrysonilia sitophila, Mucor racemosus, Paecilomyces sp. and Trichoderma viride were found in TCA-containing stoppers, while C. sitophila and Penicillium sp. were the main fungi in the stoppers devoid of TCA. Conidia were numerous close to the lenticels and present from the lateral surface through to the centre of the stoppers. Strains of Aspergillus, Mucor, Paecilomyces, Penicillium and Trichoderma isolated from TCA-containing stoppers were able to convert 2,4,6-trichlorophenol (TCP) in TCA in resting cell or growing conditions. The best yields of conversion were obtained by green fungi Paecilomyces sp. and P. chrysogenum, 17% and 20%, respectively. Chysonilia sitophila and Penicillium sp. did not produce TCA from TCP in our conditions.

  13. Variability in biofilm formation correlates with hydrophobicity and quorum sensing among Vibrio parahaemolyticus isolates from food contact surfaces and the distribution of the genes involved in biofilm formation.

    Science.gov (United States)

    Mizan, Md Furkanur Rahaman; Jahid, Iqbal Kabir; Kim, Minhui; Lee, Ki-Hoon; Kim, Tae Jo; Ha, Sang-Do

    2016-01-01

    Vibrio parahaemolyticus is one of the leading foodborne pathogens causing seafood contamination. Here, 22 V. parahaemolyticus strains were analyzed for biofilm formation to determine whether there is a correlation between biofilm formation and quorum sensing (QS), swimming motility, or hydrophobicity. The results indicate that the biofilm formation ability of V. parahaemolyticus is positively correlated with cell surface hydrophobicity, autoinducer (AI-2) production, and protease activity. Field emission scanning electron microscopy (FESEM) showed that strong-biofilm-forming strains established thick 3-D structures, whereas poor-biofilm-forming strains produced thin inconsistent biofilms. In addition, the distribution of the genes encoding pandemic clone factors, type VI secretion systems (T6SS), biofilm functions, and the type I pilus in the V. parahaemolyticus seafood isolates were examined. Biofilm-associated genes were present in almost all the strains, irrespective of other phenotypes. These results indicate that biofilm formation on/in seafood may constitute a major factor in the dissemination of V. parahaemolyticus and the ensuing diseases.

  14. One of two TolC-like proteins is involved in antibiotic resistance and biofilm formation of Actinobacillus pleuropneumoniae clinical isolate SC1516

    Directory of Open Access Journals (Sweden)

    Ying Li

    2016-10-01

    Full Text Available Actinobacillus pleuropneumoniae is the etiologic agent of porcine contagious pleuropneumonia, a significant disease that causes serious economic losses to the swine industry worldwide. Persistent infections caused by bacterial biofilms are recalcitrant to treat because of the particular drug resistance of biofilm-dwelling cells. TolC, a key component of multidrug efflux pumps, are responsible for multidrug resistance in many Gram-negative bacteria. In this study, we identified two TolC-like proteins, TolC1 and TolC2, in A. pleuropneumoniae. Deletion of tolC1, but not tolC2, caused a significant reduction in biofilm formation, as well as increased drug sensitivity of both planktonic and biofilm cells. The genetic-complementation of the tolC1 mutation restored the competent biofilm and drug resistance. Besides, biofilm formation was inhibited and drug sensitivity was increased by the addition of phenylalanine-arginine beta-naphthylamide (PAβN, a well-known efflux pump inhibitor (EPI, suggesting a role for EPI in antibacterial strategies towards drug tolerance of A. pleuropneumoniae. Taken together, TolC1 is required for biofilm formation and is a part of the multidrug resistance machinery of both planktonic and biofilm cells, which could supplement therapeutic strategies for resistant bacteria and biofilm-related infections of A. pleuropneumoniae clinical isolate SC1516.

  15. Premating isolation is determined by larval rearing substrates in cactophilic Drosophila mojavensis. IX. Host plant and population specific epicuticular hydrocarbon expression influences mate choice and sexual selection.

    Science.gov (United States)

    Havens, J A; Etges, W J

    2013-03-01

    Sexual signals in cactophilic Drosophila mojavensis include cuticular hydrocarbons (CHCs), contact pheromones that mediate female discrimination of males during courtship. CHCs, along with male courtship songs, cause premating isolation between diverged populations, and are influenced by genotype × environment interactions caused by different host cacti. CHC profiles of mated and unmated adult flies from a Baja California and a mainland Mexico population of D. mojavensis reared on two host cacti were assayed to test the hypothesis that male CHCs mediate within-population female discrimination of males. In multiple choice courtship trials, mated and unmated males differed in CHC profiles, indicating that females prefer males with particular blends of CHCs. Mated and unmated females significantly differed in CHC profiles as well. Adults in the choice trials had CHC profiles that were significantly different from those in pair-mated adults from no-choice trials revealing an influence of sexual selection. Females preferred different male CHC blends in each population, but the influence of host cactus on CHC variation was significant only in the mainland population indicating population-specific plasticity in CHCs. Different groups of CHCs mediated female choice-based sexual selection in each population suggesting that geographical and ecological divergence has the potential to promote divergence in mate communication systems.

  16. Hydrocarbon degradation and plant colonization of selected bacterial strains isolated from the rhizsophere and plant interior of Italian ryegrass and Birdsfoot trefoil

    Science.gov (United States)

    Sohail, Y.; Andria, V.; Reichenauer, T. G.; Sessitsch, A.

    2009-04-01

    Hydrocarbon-degrading strains were isolated from the rhizosphere, root and shoot interior of Italian ryegrass (Lolium multiflorum var. Taurus), Birdsfoot trefoil (Lotus corniculatus var. Leo) grown in a soil contaminated with petroleum oil. Strains were tested regarding their phylogeny and their degradation efficiency. The most efficient strains were tested regarding their suitability to be applied for phytoremediation of diesel oils. Sterilized and non-sterilized agricultural soil, with and with out compost, were spiked with diesel and used for planting Italian ryegrass and birdsfoot trefoil. Four selected strains with high degradation activities, derived from the rhizosphere and plant interior, were selected for individual inoculation. Plants were harvested at flowering stage and plant biomass and hydrocarbon degradation was determined. Furthermore, it was investigated to which extent the inoculant strains were able to survive and colonize plants. Microbial community structures were analysed by 16S rRNA and alkB gene analysis. Results showed efficient colonization by the inoculant strains and improved degradation by the application of compost combined with inoculation as well as on microbial community structures will be presented.

  17. Synthesis of a novel molecularly imprinted organic-inorganic hybrid polymer for the selective isolation and determination of fluoroquinolones in tilapia.

    Science.gov (United States)

    Yang, Xun; Wang, Ruiling; Wang, Weihua; Yan, Hongyuan; Qiu, Mande; Song, Yanxue

    2014-01-15

    A novel molecularly imprinted organic-inorganic hybrid polymer (MI-MAA/APTS) based on a dummy molecular im