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1.
Plant Signal Behav ; 14(9): 1632688, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31230564

RESUMO

Piriformospora indica, an endophytic fungus of Sebacinales, has a wide host range and promotes the performance of mono- and eudicot plants. Here, we compare the interaction of P. indica with the roots of seven host plants (Anthurium andraeanum, Arabidopsis thaliana, Brassica campestris, Lycopersicon esculentum, Oncidium orchid, Oryza sativa, and Zea mays). Microscopical analyses showed that the colonization time and the mode of hyphal invasion into the roots differ in the symbiotic interactions. Substantial differences between the species were also observed for the levels and accumulation of jasmonate (JA) and gibberellin (GA) and the transcript levels for genes involved in their syntheses. No obvious correlation could be detected between the endogenous JA and/or GA levels and the time point of root colonization in a given plant species. Our results suggest that root colonization strategies and changes in the two phytohormone levels are highly host-specific.


Assuntos
Basidiomycota/fisiologia , Especificidade de Hospedeiro , Interações Hospedeiro-Patógeno , Reguladores de Crescimento de Plantas/farmacologia , Raízes de Plantas/microbiologia , Plantas/microbiologia , Basidiomycota/efeitos dos fármacos , Vias Biossintéticas/efeitos dos fármacos , Vias Biossintéticas/genética , Ciclopentanos/metabolismo , Etilenos/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Giberelinas/metabolismo , Especificidade de Hospedeiro/efeitos dos fármacos , Interações Hospedeiro-Patógeno/efeitos dos fármacos , Interações Hospedeiro-Patógeno/genética , Oxilipinas/metabolismo , Raízes de Plantas/citologia , Raízes de Plantas/efeitos dos fármacos , Plantas/efeitos dos fármacos , Plantas/genética , Fatores de Tempo
2.
Viruses ; 11(4)2019 04 09.
Artigo em Inglês | MEDLINE | ID: mdl-30970658

RESUMO

As an invasive weed, Mikaniamicrantha Kunth has caused serious damage to natural forest ecosystems in South China in recent years. Mikania micrantha wilt virus (MMWV), an isolate of the Gentian mosaic virus (GeMV), is transmitted by Myzuspersicae (Sulzer) in a non-persistent manner and can effectively inhibit the growth of M. micrantha. To explore the MMWV-M. micrantha-M. persicae interaction and its impact on the invasion of M. micrantha, volatile compounds (VOCs) emitted from healthy, mock-inoculated, and MMWV-infected plants were collected, and effects on host preference of the apterous and alate aphids were assessed with Y-shaped olfactometers. Gas chromatography-mass spectrometry (GC-MS) analysis indicated that MMWV infection changed the VOC profiles, rendering plants more attractive to aphids. Clip-cages were used to document the population growth rate of M.persicae fed on healthy, mock-inoculated, or MMWV-infected plants. Compared to those reared on healthy plants, the population growth of M. persicae drastically decreased on the MMWV-infected plants. Plant host choice tests based on visual and contact cues were also conducted using alate M.persicae. Interestingly, the initial attractiveness of MMWV-infected plants diminished, and more alate M. persicae moved to healthy plants. Taken together, MMWV appeared to be able to manipulate its plant host to first attract insect vectors to infected plants but then repel viruliferous vectors to promote its own dispersal. Its potential application for invasive weed management is discussed.


Assuntos
Afídeos/fisiologia , Fabavirus/crescimento & desenvolvimento , Comportamento Alimentar/efeitos dos fármacos , Interações entre Hospedeiro e Microrganismos , Especificidade de Hospedeiro/efeitos dos fármacos , Mikania/virologia , Feromônios/metabolismo , Animais , Afídeos/efeitos dos fármacos , China , Insetos Vetores/efeitos dos fármacos , Insetos Vetores/fisiologia , Doenças das Plantas/virologia , Compostos Orgânicos Voláteis/metabolismo
3.
Sci Rep ; 8(1): 12772, 2018 08 24.
Artigo em Inglês | MEDLINE | ID: mdl-30143740

RESUMO

In the current report, we describe the identification of three genetically distinct groups of prophages integrated into three different chromosomal sites of human gut-associated Bifidobacterium breve and Bifidobacterium longum strains. These bifidobacterial prophages are distantly related to temperate actinobacteriophages of several hosts. Some prophages, integrated within the dnaJ2 gene, are competent for induction, excision, replication, assembly and lysis, suggesting that they are fully functional and can generate infectious particles, even though permissive hosts have not yet been identified. Interestingly, several of these phages harbor a putative phase variation shufflon (the Rin system) that generates variation of the tail-associated receptor binding protein (RBP). Unlike the analogous coliphage-associated shufflon Min, or simpler Cin and Gin inversion systems, Rin is predicted to use a tyrosine recombinase to promote inversion, the first reported phage-encoded tyrosine-family DNA invertase. The identification of bifidobacterial prophages with RBP diversification systems that are competent for assembly and lysis, yet fail to propagate lytically under laboratory conditions, suggests dynamic evolution of bifidobacteria and their phages in the human gut.


Assuntos
Bifidobacterium/virologia , Microbioma Gastrointestinal , Prófagos/fisiologia , Sítios de Ligação Microbiológicos/genética , Sequência de Bases , Bifidobacterium/efeitos dos fármacos , Evolução Biológica , Microbioma Gastrointestinal/efeitos dos fármacos , Genoma Viral , Especificidade de Hospedeiro/efeitos dos fármacos , Especificidade de Hospedeiro/genética , Humanos , Mitomicina/farmacologia , Prófagos/efeitos dos fármacos , Prófagos/genética , Prófagos/ultraestrutura , Vírion/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos
4.
Nat Microbiol ; 2(12): 1592-1599, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28993610

RESUMO

The evolution of virulence traits is central for the emergence or re-emergence of microbial pathogens and for their adaptation to a specific host 1-5 . Typhoid toxin is an essential virulence factor of the human-adapted bacterial pathogen Salmonella Typhi 6,7 , the cause of typhoid fever in humans 8-12 . Typhoid toxin has a unique A2B5 architecture with two covalently linked enzymatic 'A' subunits, PltA and CdtB, associated with a homopentameric 'B' subunit made up of PltB, which has binding specificity for the N-acetylneuraminic acid (Neu5Ac) sialoglycans 6,13 prominently present in humans 14 . Here, we examine the functional and structural relationship between typhoid toxin and ArtAB, an evolutionarily related AB5 toxin encoded by the broad-host Salmonella Typhimurium 15 . We find that ArtA and ArtB, homologues of PltA and PltB, can form a functional complex with the typhoid toxin CdtB subunit after substitution of a single amino acid in ArtA, while ArtB can form a functional complex with wild-type PltA and CdtB. We also found that, after addition of a single-terminal Cys residue, a CdtB homologue from cytolethal distending toxin can form a functional complex with ArtA and ArtB. In line with the broad host specificity of S. Typhimurium, we found that ArtB binds human glycans, terminated in N-acetylneuraminic acid, as well as glycans terminated in N-glycolylneuraminic acid (Neu5Gc), which are expressed in most other mammals 14 . The atomic structure of ArtB bound to its receptor shows the presence of an additional glycan-binding site, which broadens its binding specificity. Despite equivalent toxicity in vitro, we found that the ArtB/PltA/CdtB chimaeric toxin exhibits reduced lethality in an animal model, indicating that the host specialization of typhoid toxin has optimized its targeting mechanisms to the human host. This is a remarkable example of a toxin evolving to broaden its enzymatic activities and adapt to a specific host.


Assuntos
Adaptação Fisiológica , Endotoxinas/toxicidade , Especificidade de Hospedeiro/efeitos dos fármacos , Especificidade de Hospedeiro/fisiologia , Salmonella typhi/química , Sequência de Aminoácidos , Animais , Proteínas de Bactérias/metabolismo , Toxinas Bacterianas/metabolismo , Sítios de Ligação , Linhagem Celular , Cristalografia por Raios X , Glicômica , Células HEK293 , Humanos , Masculino , Camundongos , Modelos Moleculares , Ácido N-Acetilneuramínico/química , Ácido N-Acetilneuramínico/metabolismo , Ácidos Neuramínicos/química , Ácidos Neuramínicos/metabolismo , Polissacarídeos/metabolismo , Salmonella typhi/patogenicidade , Fatores de Transcrição , Febre Tifoide/microbiologia , Fatores de Virulência
5.
Food Funct ; 7(8): 3531-8, 2016 Aug 10.
Artigo em Inglês | MEDLINE | ID: mdl-27435508

RESUMO

Adhesion to the intestinal epithelium is considered an important feature of probiotic bacteria, which may increase their persistence in the intestine, allowing them to exert their beneficial health effect or promote the colonisation process. However, this feature might be largely dependent on the host specificity or diet. In the present study, we investigated the effect of selected milks and milk protein fractions on the ability of selected lactobacilli to adhere to the cells of an intestinal model based on co-culture Caco-2/HT29-MTX cell lines. Most milk digesta did not significantly affect bacterial adhesion except for UHT-treated milk and sheep milk. The presence of UHT-treated milk digesta reduced the adhesion of Lactobacillus gasseri R by 61% but not that of Lactobacillus casei FMP. However, sheep milk significantly increased the adherence of L. casei FMP (P < 0.05) but not of L. gasseri R. Among the protein fractions, rennet casein (RCN) and bovine serum albumin (BSA) showed reproducible patterns and strain-specific effects on bacterial adherence. While RCN reduced the adherence of L. gasseri R to <50% compared to the control, it did not have a significant effect on L. casei FMP. In contrast, BSA reduced L. casei FMP adherence to a higher extent than that of L. gasseri R. Whey protein (WH) tended to increase the adherence of both strains by 130%-180%. Recently, interactions between the host diet and its microbiota have attracted considerable interest. Our results may explain one of the aspects of the role of milk in the development of microbiota or support of probiotic supplements. Based on our data, we conclude that the persistence of probiotic strains supplemented as part of dairy food or constitutional microbiota in the gut might be affected negatively or positively by the food matrix through complex strain or concentration dependent effects.


Assuntos
Aderência Bacteriana/efeitos dos fármacos , Lacticaseibacillus casei/efeitos dos fármacos , Lactobacillus gasseri/efeitos dos fármacos , Proteínas do Leite/farmacologia , Leite/química , Leite/microbiologia , Animais , Células CACO-2 , Sobrevivência Celular/efeitos dos fármacos , Técnicas de Cocultura , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Epitélio/efeitos dos fármacos , Epitélio/metabolismo , Células HT29 , Especificidade de Hospedeiro/efeitos dos fármacos , Humanos , Mucosa Intestinal/metabolismo , Intestinos/citologia , Intestinos/efeitos dos fármacos , Ovinos
6.
J Chem Ecol ; 42(5): 394-403, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-27240622

RESUMO

The chemical composition of plant surfaces plays a role in selection of host plants by herbivorous insects. Once the insect reaches the plant, these cues determine host acceptance. Laboratory studies have shown that the stem borer Busseola fusca (Lepidoptera: Noctuidae), an important pest of sorghum and maize in sub-Saharan Africa, is able to differentiate between host and non-host plant species. However, no information is available on the cues used by this insect to seek and accept the host plant. Thus, the role of surface phytochemical stimuli on host selection and oviposition by B. fusca was studied in the laboratory using two host plants, sorghum, Sorghum bicolor, and maize, Zea mays, and one non-host plant, Napier grass, Pennisetum purpureum. The numbers of eggs and egg masses deposited on the three plant species were compared first under no-choice and choice conditions. In both cases, more eggs and egg masses were laid on maize and sorghum than on the non-host. Artificial surrogate stems treated with a water or chloroform surface extract of each plant were then compared with surrogate stems treated with, respectively, water or chloroform as controls, under similar conditions. Surrogate stems treated with plant water extracts did not show an increase in oviposition when compared to controls, indicating that the major compounds in these extracts, i.e., simple sugars and free amino acids, are not significantly responsible for the oviposition preference. By contrast, a chloroform extract of sorghum enhanced oviposition on the surrogate stems compared to the control, while those of maize and Napier grass showed no significant effects. Analysis of the chloroform extract of sorghum showed higher amounts of α-amyrin, ß-amyrin, and n-nonacosane compared to those of maize and Napier grass. A blend of the three chemicals significantly increased oviposition compared to the chloroform-treated control, indicating that these compounds are part of the surface chemical signature of the plant responsible for host recognition and oviposition by B. fusca.


Assuntos
Especificidade de Hospedeiro/efeitos dos fármacos , Mariposas/efeitos dos fármacos , Mariposas/fisiologia , Oviposição/efeitos dos fármacos , Extratos Vegetais/farmacologia , Animais , Bioensaio , Clorofórmio/química , Grão Comestível , Feminino , Herbivoria/efeitos dos fármacos , Extratos Vegetais/análise , Propriedades de Superfície , Água/química
7.
J Chem Ecol ; 42(5): 357-67, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-27193763

RESUMO

Plant pathogens can manipulate the odor of their host; the odor of an infected plant is often attractive to the plant pathogen vector. It has been suggested that this odor-mediated manipulation attracts vectors and may contribute to spread of disease; however, this requires further broad demonstration among vector-pathogen systems. In addition, disruption of this indirect chemical communication between the pathogen and the vector has not been attempted. We present a model that demonstrates how a phytophathogen (Candidatus Liberibacter asiaticus) can increase its spread by indirectly manipulating the behavior of its vector (Asian citrus psyllid, Diaphorina citri Kuwayama). The model indicates that when vectors are attracted to pathogen-infected hosts, the proportion of infected vectors increases, as well as, the proportion of infected hosts. Additionally, the peak of infected host populations occurs earlier as compared with controls. These changes in disease dynamics were more important during scenarios with higher vector mortality. Subsequently, we conducted a series of experiments to disrupt the behavior of the Asian citrus psyllid. To do so, we exposed the vector to methyl salicylate, the major compound released following host infection with the pathogen. We observed that during exposure or after pre-exposure to methyl salicylate, the host preference can be altered; indeed, the Asian citrus psyllids were unable to select infected hosts over uninfected counterparts. We suggest mechanisms to explain these interactions and potential applications of disrupting herbivore host preference with plant volatiles for sustainable management of insect vectors.


Assuntos
Citrus/microbiologia , Hemípteros/efeitos dos fármacos , Especificidade de Hospedeiro/efeitos dos fármacos , Insetos Vetores/efeitos dos fármacos , Rhizobiaceae/fisiologia , Compostos Orgânicos Voláteis/farmacologia , Animais , Citrus/metabolismo , Relação Dose-Resposta a Droga , Hemípteros/fisiologia , Insetos Vetores/fisiologia , Modelos Estatísticos , Feromônios/metabolismo , Feromônios/farmacologia , Doenças das Plantas/microbiologia , Rhizobiaceae/efeitos dos fármacos , Salicilatos/metabolismo , Salicilatos/farmacologia , Compostos Orgânicos Voláteis/metabolismo
8.
Sci Rep ; 5: 17257, 2015 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-26607832

RESUMO

The increasing emergence of multi-drug resistant streptococci poses a serious threat to public health worldwide. Bacteriophage lysins are promising alternatives to antibiotics; however, their narrow lytic spectrum restricted to closely related species is a central shortcoming to their translational development. Here, we describe an efficient method for rapid screening of engineered chimeric lysins and report a unique "chimeolysin", ClyR, with robust activity and an extended-spectrum streptococcal host range against most streptococcal species, including S. pyogenes, S. agalactiae, S. dysgalactiae, S. equi, S. mutans, S. pneumoniae, S. suis and S. uberis, as well as representative enterococcal and staphylococcal species (including MRSA and VISA). ClyR is the first lysin that demonstrates activity against the dominant dental caries-causing pathogen as well as the first lysin that kills all four of the bovine mastitis-causing pathogens. This study demonstrates the success of the screening method resulting in a powerful lysin with potential for treating most streptococcal associated infections.


Assuntos
Bacteriólise/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos/métodos , Especificidade de Hospedeiro/efeitos dos fármacos , Proteínas Recombinantes/farmacologia , Streptococcus/fisiologia , Animais , Bovinos , Modelos Animais de Doenças , Escherichia coli , Feminino , Camundongos Endogâmicos BALB C , Leite/química , Infecções Estreptocócicas , Streptococcus/efeitos dos fármacos
9.
PLoS One ; 9(2): e88370, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24520377

RESUMO

The evolution of cactophily in the genus Drosophila was a major ecological transition involving over a hundred species in the Americas that acquired the capacity to cope with a variety of toxic metabolites evolved as feeding deterrents in Cactaceae. D. buzzatii and D. koepferae are sibling cactophilic species in the D. repleta group. The former is mainly associated with the relatively toxic-free habitat offered by prickly pears (Opuntia sulphurea) and the latter has evolved the ability to use columnar cacti of the genera Trichocereus and Cereus that contain an array of alkaloid secondary compounds. We assessed the effects of cactus alkaloids on fitness-related traits and evaluated the ability of D. buzzatii and D. koepferae to exploit an artificial novel toxic host. Larvae of both species were raised in laboratory culture media to which we added increasing doses of an alkaloid fraction extracted from the columnar cactus T. terschekii. In addition, we evaluated performance on an artificial novel host by rearing larvae in a seminatural medium that combined the nutritional quality of O. sulphurea plus amounts of alkaloids found in fresh T. terschekii. Performance scores in each rearing treatment were calculated using an index that took into account viability, developmental time, and adult body size. Only D. buzzatii suffered the effects of increasing doses of alkaloids and the artificial host impaired viability in D. koepferae, but did not affect performance in D. buzzatii. These results provide the first direct evidence that alkaloids are key determinants of host plant use in these species. However, the results regarding the artificial novel host suggest that the effects of alkaloids on performance are not straightforward as D. koepferae was heavily affected. We discuss these results in the light of patterns of host plan evolution in the Drosophila repleta group.


Assuntos
Adaptação Fisiológica/efeitos dos fármacos , Alcaloides/toxicidade , Cactaceae/química , Drosophila/efeitos dos fármacos , Drosophila/fisiologia , Especificidade de Hospedeiro/efeitos dos fármacos , Animais , Análise de Regressão
10.
Insect Sci ; 21(1): 47-55, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23956152

RESUMO

Jasmonate- and salicylate-mediated signaling pathways play significant roles in induced plant defenses, but there is no sufficient evidence for their roles in monocots against aphids. We exogenously applied methyl jasmonate (MeJA) and salicylic acid (SA) on wheat seedlings and examined biochemical responses in wheat and effects on the grain aphid, Sitobion avenae (Fab.). Application of MeJA significantly increased levels of wheat's polyphenol oxidase, peroxidase and proteinase inhibitor 1, 2 and 6 days after treatment. In two-choice tests, adult aphids preferred control wheat leaves to MeJA- or SA-treated leaves. Electrical penetration graph (EPG) recordings of aphid probing behavior revealed that on MeJA-treated plants, the duration of aphid's first probe was significantly shorter and number of probes was significantly higher than those on control plants. Also total duration of probing on MeJA-treated plants was significantly shorter than on control plants. Total duration of salivation period on SA-treated plants was significantly longer, while mean phloem ingestion period was significantly shorter than on control plants. However, no significant difference in total duration of phloem sap ingestion period was observed among treatments. The EPG data suggest that MeJA-dependent resistance factors might be due to feeding deterrents in mesophyll, whereas the SA-mediated resistance may be phloem-based. We did not observe any significant difference of MeJA and SA application on aphid development, daily fecundity, intrinsic growth rate and population growth. The results indicate that both MeJA- and SA-induced defenses in wheat deterred S. avenae colonization processes and feeding behavior, but had no significant effects on its performance.


Assuntos
Afídeos/fisiologia , Ciclopentanos/farmacologia , Oxilipinas/farmacologia , Doenças das Plantas/parasitologia , Ácido Salicílico/farmacologia , Triticum/imunologia , Triticum/parasitologia , Animais , Afídeos/efeitos dos fármacos , Comportamento Alimentar/efeitos dos fármacos , Feminino , Especificidade de Hospedeiro/efeitos dos fármacos , Masculino , Doenças das Plantas/imunologia , Folhas de Planta/imunologia , Folhas de Planta/parasitologia , Triticum/efeitos dos fármacos
11.
Nature ; 499(7457): 219-22, 2013 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-23748443

RESUMO

The mammalian gut ecosystem has considerable influence on host physiology, but the mechanisms that sustain this complex environment in the face of different stresses remain obscure. Perturbations to the gut ecosystem, such as through antibiotic treatment or diet, are at present interpreted at the level of bacterial phylogeny. Less is known about the contributions of the abundant population of phages to this ecological network. Here we explore the phageome as a potential genetic reservoir for bacterial adaptation by sequencing murine faecal phage populations following antibiotic perturbation. We show that antibiotic treatment leads to the enrichment of phage-encoded genes that confer resistance via disparate mechanisms to the administered drug, as well as genes that confer resistance to antibiotics unrelated to the administered drug, and we demonstrate experimentally that phages from treated mice provide aerobically cultured naive microbiota with increased resistance. Systems-wide analyses uncovered post-treatment phage-encoded processes related to host colonization and growth adaptation, indicating that the phageome becomes broadly enriched for functionally beneficial genes under stress-related conditions. We also show that antibiotic treatment expands the interactions between phage and bacterial species, leading to a more highly connected phage-bacterial network for gene exchange. Our work implicates the phageome in the emergence of multidrug resistance, and indicates that the adaptive capacity of the phageome may represent a community-based mechanism for protecting the gut microflora, preserving its functional robustness during antibiotic stress.


Assuntos
Antibacterianos/farmacologia , Bacteriófagos/efeitos dos fármacos , Resistência Microbiana a Medicamentos/efeitos dos fármacos , Fezes/microbiologia , Fezes/virologia , Genoma Viral/genética , Metagenoma/genética , Aerobiose , Ampicilina/farmacologia , Animais , Bacteriófagos/genética , Bacteriófagos/isolamento & purificação , Ciprofloxacina/farmacologia , Resistência Microbiana a Medicamentos/genética , Feminino , Transferência Genética Horizontal/efeitos dos fármacos , Transferência Genética Horizontal/genética , Genes Virais/efeitos dos fármacos , Genes Virais/genética , Especificidade de Hospedeiro/efeitos dos fármacos , Metagenoma/efeitos dos fármacos , Camundongos , Simbiose/efeitos dos fármacos , Simbiose/genética
12.
Nature ; 498(7455): 487-91, 2013 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-23719379

RESUMO

Female mosquitoes of some species are generalists and will blood-feed on a variety of vertebrate hosts, whereas others display marked host preference. Anopheles gambiae and Aedes aegypti have evolved a strong preference for humans, making them dangerously efficient vectors of malaria and Dengue haemorrhagic fever. Specific host odours probably drive this strong preference because other attractive cues, including body heat and exhaled carbon dioxide (CO2), are common to all warm-blooded hosts. Insects sense odours via several chemosensory receptor families, including the odorant receptors (ORs), membrane proteins that form heteromeric odour-gated ion channels comprising a variable ligand-selective subunit and an obligate co-receptor called Orco (ref. 6). Here we use zinc-finger nucleases to generate targeted mutations in the orco gene of A. aegypti to examine the contribution of Orco and the odorant receptor pathway to mosquito host selection and sensitivity to the insect repellent DEET (N,N-diethyl-meta-toluamide). orco mutant olfactory sensory neurons have greatly reduced spontaneous activity and lack odour-evoked responses. Behaviourally, orco mutant mosquitoes have severely reduced attraction to honey, an odour cue related to floral nectar, and do not respond to human scent in the absence of CO2. However, in the presence of CO2, female orco mutant mosquitoes retain strong attraction to both human and animal hosts, but no longer strongly prefer humans. orco mutant females are attracted to human hosts even in the presence of DEET, but are repelled upon contact, indicating that olfactory- and contact-mediated effects of DEET are mechanistically distinct. We conclude that the odorant receptor pathway is crucial for an anthropophilic vector mosquito to discriminate human from non-human hosts and to be effectively repelled by volatile DEET.


Assuntos
Aedes/genética , Aedes/fisiologia , DEET/farmacologia , Genes de Insetos/genética , Especificidade de Hospedeiro/genética , Repelentes de Insetos/farmacologia , Mutação/genética , Aedes/efeitos dos fármacos , Sequência de Aminoácidos , Animais , Sequência de Bases , DEET/administração & dosagem , Resistência a Medicamentos/efeitos dos fármacos , Feminino , Mel , Especificidade de Hospedeiro/efeitos dos fármacos , Humanos , Repelentes de Insetos/administração & dosagem , Masculino , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Neurônios/citologia , Neurônios/efeitos dos fármacos , Odorantes/análise , Condutos Olfatórios/citologia , Condutos Olfatórios/efeitos dos fármacos , Volatilização
13.
New Phytol ; 198(4): 1017-1022, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23550729

RESUMO

· Besides photosynthates, dodder (Cuscuta spp.) acquires phloem-mobile proteins from host; however, whether this could mediate inter-species phenotype transfer was not demonstrated. Specifically, we test whether phosphinothricin acetyl transferase (PAT) that confers host plant glufosinate herbicide tolerance traffics and functions inter-specifically. · Dodder tendrils excised from hosts can grow in vitro for weeks or resume in vivo by parasitizing new hosts. The level of PAT in in vivo and in vitro dodder tendrils was quantified by enzyme-linked immunosorbent assay. The glufosinate sensitivity was examined by dipping the distal end of in vivo and in vitro tendrils, growing on or excised from LibertyLink (LL; PAT-transgenic and glufosinate tolerant) and conventional (CN; glufosinate sensitive) soybean hosts, into glufosinate solutions for 5 s. After in vitro tendrils excised from LL hosts reparasitized new CN and LL hosts, the PAT level and the glufosinate sensitivity were also examined. · When growing on LL host, dodder tolerated glufosinate and contained PAT at a level of 0.3% of that encountered in LL soybean leaf. After PAT was largely degraded in dodders, they became glufosinate sensitive. PAT mRNA was not detected by reverse transcription PCR in dodders. · In conclusion, the results indicated that PAT inter-species trafficking confers dodder glufosinate tolerance.


Assuntos
Adaptação Fisiológica/efeitos dos fármacos , Cuscuta/efeitos dos fármacos , Cuscuta/fisiologia , Herbicidas/toxicidade , Especificidade de Hospedeiro/efeitos dos fármacos , Proteínas de Plantas/metabolismo , Característica Quantitativa Herdável , Acetiltransferases/genética , Acetiltransferases/metabolismo , Adaptação Fisiológica/genética , Aminobutiratos/toxicidade , Cuscuta/enzimologia , Cuscuta/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Plantas Geneticamente Modificadas , Transporte Proteico/efeitos dos fármacos , Proteólise/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Glycine max , Especificidade da Espécie
14.
PLoS One ; 7(2): e31839, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22363747

RESUMO

The role of the NS1 protein in modulating influenza A virulence and host range was assessed by adapting A/Hong Kong/1/1968 (H3N2) (HK-wt) to increased virulence in the mouse. Sequencing the NS genome segment of mouse-adapted variants revealed 11 mutations in the NS1 gene and 4 in the overlapping NEP gene. Using the HK-wt virus and reverse genetics to incorporate mutant NS gene segments, we demonstrated that all NS1 mutations were adaptive and enhanced virus replication (up to 100 fold) in mouse cells and/or lungs. All but one NS1 mutant was associated with increased virulence measured by survival and weight loss in the mouse. Ten of twelve NS1 mutants significantly enhanced IFN-ß antagonism to reduce the level of IFN ß production relative to HK-wt in infected mouse lungs at 1 day post infection, where 9 mutants induced viral yields in the lung that were equivalent to or significantly greater than HK-wt (up to 16 fold increase). Eight of 12 NS1 mutants had reduced or lost the ability to bind the 30 kDa cleavage and polyadenylation specificity factor (CPSF30) thus demonstrating a lack of correlation with reduced IFN ß production. Mutant NS1 genes resulted in increased viral mRNA transcription (10 of 12 mutants), and protein production (6 of 12 mutants) in mouse cells. Increased transcription activity was demonstrated in the influenza mini-genome assay for 7 of 11 NS1 mutants. Although we have shown gain-of-function properties for all mutant NS genes, the contribution of the NEP mutations to phenotypic changes remains to be assessed. This study demonstrates that NS1 is a multifunctional virulence factor subject to adaptive evolution.


Assuntos
Adaptação Biológica/genética , Especificidade de Hospedeiro/genética , Vírus da Influenza A/genética , Influenza Humana/virologia , Mutação/genética , Seleção Genética , Proteínas não Estruturais Virais/genética , Adaptação Biológica/efeitos dos fármacos , Animais , Bioensaio , Regulação Viral da Expressão Gênica/efeitos dos fármacos , Meia-Vida , Especificidade de Hospedeiro/efeitos dos fármacos , Humanos , Vírus da Influenza A/efeitos dos fármacos , Vírus da Influenza A/crescimento & desenvolvimento , Vírus da Influenza A/patogenicidade , Interferon beta/biossíntese , Interferon beta/farmacologia , Pulmão/efeitos dos fármacos , Pulmão/patologia , Pulmão/virologia , Camundongos , Dados de Sequência Molecular , Proteínas Mutantes/metabolismo , Infecções por Orthomyxoviridae/patologia , Infecções por Orthomyxoviridae/virologia , Ligação Proteica/efeitos dos fármacos , Biossíntese de Proteínas/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Recombinação Genética/genética , Virulência/efeitos dos fármacos
15.
Biotechniques ; 50(5): 303-9, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21548892

RESUMO

Vaccinia virus has been a powerful tool in molecular biology and vaccine development. The relative ease of inserting and expressing foreign genes combined with its broad host range has made it an attractive antigen delivery system against many heterologous diseases. Many different approaches have been developed to isolate recombinant vaccinia virus generated from homologous recombination; however, most are time-consuming, often requiring a series of passages or specific cell lines. Herein we introduce a rapid method for isolating recombinants using the antibiotic coumermycin and the interferon-associated PKR pathway to select for vaccinia virus recombinants. This method uses a negative selection marker in the form of a fusion protein, GyrB-PKR, consisting of the coumermycin dimerization domain of Escherichia coli gyrase subunit B fused to the catalytic domain of human PKR. Coumermycin-dependent dimerization of this protein results in activation of PKR and the phosphorylation of translation initiation factor, eIF2α. Phosphorylation of this factor leads to an inhibition of protein synthesis, and an inhibition of virus replication. In the presence of coumermycin, recombinants are isolated due to the loss of this coumermycin-sensitive gene by homologous recombination. We demonstrate that this method of selection is highly efficient and requires limited rounds of enrichment to isolate recombinant virus.


Assuntos
Fator de Iniciação 2 em Eucariotos/química , Vaccinia virus/isolamento & purificação , Aminocumarinas/química , Aminocumarinas/farmacologia , Animais , Linhagem Celular , Cricetinae , Dimerização , Escherichia coli/genética , Escherichia coli/metabolismo , Vetores Genéticos , Especificidade de Hospedeiro/efeitos dos fármacos , Inibidores da Síntese de Proteínas/química , Inibidores da Síntese de Proteínas/farmacologia , Coelhos , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/efeitos dos fármacos , Proteínas Recombinantes/isolamento & purificação , Recombinação Genética/efeitos dos fármacos , Vaccinia virus/efeitos dos fármacos , Vaccinia virus/crescimento & desenvolvimento , Replicação Viral/efeitos dos fármacos , Replicação Viral/genética
16.
Parasitol Res ; 109(4): 1143-9, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21479576

RESUMO

Malaria is one of the most lethal parasitic infections in the world. The lethality of the parasite depends on the rate of multiplication of the parasite within host erythrocytes. Different strains of the malaria parasite often respond in a different way to the same strain of mice or vice versa. In the present study, we investigated the course of infection of the arteether-sensitive and arteether-resistant Plasmodium vinckei parasites in Swiss albino AKR (inbred) and AJ (outbred) mice. The higher parasite burden and mortality were observed in the sensitive parasite-infected mice, whereas the infection with the resistant parasite was non-lethal. Resistant parasite-infected mice developed a moderate level of parasitemia that decreased gradually throughout the infection. The microscopic examination suggests that the resistant parasite invades reticulocytes more efficiently than normocytes, regardless of the mouse strain examined. Since the reticulocytes are rare in blood circulation, it limits the increase in parasite proliferations, while arteether-sensitive parasites can invade both mature normocytes and reticulocytes, resulting in the mortality of the mice. However, treatment with phenylhydrazine in Swiss mice results in reticulocytosis, which transforms the non-lethal resistant parasites to produce lethal infections. Our findings demonstrate that the characteristic response during infections with the arteether-resistant strain is dependent on the availability of reticulocytes in peripheral blood circulation. We can use this model for identifying the interaction between host and artemisinin derivative-resistant parasites.


Assuntos
Eritrócitos/parasitologia , Malária/parasitologia , Plasmodium/patogenicidade , Reticulócitos/parasitologia , Doenças dos Roedores/parasitologia , Animais , Artemisininas/farmacologia , Corantes Azur , Resistência a Medicamentos , Eritrócitos/citologia , Especificidade de Hospedeiro/efeitos dos fármacos , Interações Hospedeiro-Parasita/efeitos dos fármacos , Malária/tratamento farmacológico , Camundongos , Microscopia , Parasitemia , Fenil-Hidrazinas/administração & dosagem , Fenil-Hidrazinas/efeitos adversos , Plasmodium/efeitos dos fármacos , Plasmodium/fisiologia , Contagem de Reticulócitos , Reticulócitos/citologia , Doenças dos Roedores/tratamento farmacológico , Especificidade da Espécie
17.
PLoS One ; 6(3): e18072, 2011 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-21464894

RESUMO

BACKGROUND: Gammaretroviruses and gammaretroviral vectors, in contrast to lentiviruses and lentiviral vectors, are reported to be restricted in their ability to infect growth-arrested cells. The block to this restriction has never been clearly defined. The original assessment of the inability of gammaretroviruses and gammaretroviral vectors to infect growth-arrested cells was carried out using established cell lines that had been growth-arrested by chemical means, and has been generalized to neurons, which are post-mitotic. We re-examined the capability of gammaretroviruses and their derived vectors to efficiently infect terminally differentiated neuroendocrine cells and primary cortical neurons, a target of both experimental and therapeutic interest. METHODOLOGY/PRINCIPAL FINDINGS: Using GFP expression as a marker for infection, we determined that both growth-arrested (NGF-differentiated) rat pheochromocytoma cells (PC12 cells) and primary rat cortical neurons could be efficiently transduced, and maintained long-term protein expression, after exposure to murine leukemia virus (MLV) and MLV-based retroviral vectors. Terminally differentiated PC12 cells transduced with a gammaretroviral vector encoding the anti-apoptotic protein Bcl-xL were protected from cell death induced by withdrawal of nerve growth factor (NGF), demonstrating gammaretroviral vector-mediated delivery and expression of genes at levels sufficient for therapeutic effect in non-dividing cells. Post-mitotic rat cortical neurons were also shown to be susceptible to transduction by murine replication-competent gammaretroviruses and gammaretroviral vectors. CONCLUSIONS/SIGNIFICANCE: These findings suggest that the host range of gammaretroviruses includes post-mitotic and other growth-arrested cells in mammals, and have implications for re-direction of gammaretroviral gene therapy to neurological disease.


Assuntos
Gammaretrovirus/genética , Vetores Genéticos/genética , Especificidade de Hospedeiro/fisiologia , Mitose , Neurônios/citologia , Neurônios/virologia , Animais , Morte Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Córtex Cerebral/citologia , DNA/metabolismo , Fase G2/efeitos dos fármacos , Gammaretrovirus/efeitos dos fármacos , Proteínas de Fluorescência Verde/metabolismo , Especificidade de Hospedeiro/efeitos dos fármacos , Lentivirus/efeitos dos fármacos , Lentivirus/genética , Mitose/efeitos dos fármacos , Fator de Crescimento Neural/farmacologia , Neurônios/efeitos dos fármacos , Células PC12 , Ratos , Fase S/efeitos dos fármacos , Soro , Transdução Genética , Proteína bcl-X/metabolismo
18.
Mol Plant Pathol ; 12(2): 123-35, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21199563

RESUMO

UNLABELLED: Elsinoë fawcettii and E. australis are important pathogens of citrus. Both species are known to produce red or orange pigments, called elsinochrome. Elsinochrome is a nonhost-selective phytotoxin and is required for full fungal virulence and lesion formation. This article discusses the taxonomy, epidemiology, genetics and pathology of the pathogens. It also provides a perspective on the cellular toxicity, biosynthetic regulation and pathological role of elsinochrome phytotoxin. TAXONOMY: Elsinoë fawcettii (anamorph: Sphaceloma fawcettii) and E. australis (anamorph: S. australis) are classified in the Phylum Ascomycota, Class Dothideomycetes, Order Myriangiales and Family Elsinoaceae. HOST RANGE: Elsinoë fawcettii causes citrus scab (formerly sour orange scab and common scab) on various species and hybrids in the Rutaceae family worldwide, whereas E. australis causes sweet orange scab, primarily on sweet orange and some mandarins, and has a limited geographical distribution. DISEASE SYMPTOMS: Citrus tissues infested with Elsinoë often display erumpent scab pustules with a warty appearance. TOXIN PRODUCTION: Elsinochrome and many perylenequinone-containing phytotoxins of fungal origin are grouped as photosensitizing compounds that are able to absorb light energy, react with oxygen molecules and produce reactive oxygen species, such as superoxide and singlet oxygen. Elsinochrome has been documented to cause peroxidation of cell membranes and to induce rapid electrolyte leakage from citrus tissues. Elsinochrome biosynthesis and conidiation are coordinately regulated in E. fawcettii, and the environmental and physiological inducers commonly involved in both processes have begun to be elucidated.


Assuntos
Ascomicetos/fisiologia , Citrus/microbiologia , Doenças das Plantas/microbiologia , Antifúngicos/farmacologia , Ascomicetos/classificação , Ascomicetos/genética , Ascomicetos/patogenicidade , Citrus/efeitos dos fármacos , Especificidade de Hospedeiro/efeitos dos fármacos , Perileno/análogos & derivados , Perileno/química , Perileno/toxicidade , Doenças das Plantas/estatística & dados numéricos , Quinonas/química , Quinonas/toxicidade
19.
Vestn Ross Akad Med Nauk ; (12): 23-6, 2010.
Artigo em Russo | MEDLINE | ID: mdl-21395059

RESUMO

The limited use of mathematical simulation in epidemiology is due not only to the difficulty of monitoring the epidemic process and identifying its parameters but also to the application of oversimplified models. It is shown that realistic reproduction of actual morbidity dynamics requires taking into account heterogeneity and finiteness of the population and seasonal character of pathogen transmission mechanism.


Assuntos
Controle de Doenças Transmissíveis/métodos , Surtos de Doenças/prevenção & controle , Monitoramento Ambiental/métodos , Modelos Teóricos , Controle de Doenças Transmissíveis/organização & administração , Demografia/estatística & dados numéricos , Surtos de Doenças/estatística & dados numéricos , Monitoramento Ambiental/estatística & dados numéricos , Especificidade de Hospedeiro/efeitos dos fármacos , Humanos , Vacinação em Massa
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