RESUMO
Genomic technologies are having an increasing impact across medicine, including primary care. To enable their wider adoption and realize their potential, education of primary health-care practitioners will be required. To enable the development of such resources, understanding where GPs currently access genomic information is needed. One-hundred fifty-nine UK GPs completed the survey in response to an open invitation, between September 2017 and September 2018. Questions were in response to 4 clinical genomic scenarios, with further questions exploring resources used for rare disease patients, direct-to-consumer genetic testing and collecting a family history. Respondents were most commonly GP principals (independent GPs who own their clinic) (64.8%), aged 35-49 years (54%), worked as a GP for more than 15 years (44%) and practiced within suburban locations (typically wealthier) (50.3%). The most popular 'just in time' education source for all clinical genomic scenarios were online primary care focussed resources with general Internet search engines also popular. For genomic continuous medical education, over 70% of respondents preferred online learning. Considering specific scenarios, local guidelines were a popular resource for the familial breast cancer scenario. A large proportion (41%) had not heard of Genomics England's 100,000 genome project. Few respondents (4%) would access rare disease specific Internet resources (Orphanet, OMIM). Twenty-five percent of respondents were unsure how to respond to a direct-to-consumer commercial genetic test query, with 41% forwarding such queries to local genetic services. GPs require concise, relevant, primary care focussed resources in trusted and familiar online repositories of information. Inadequate genetic education of GPs could increase burden on local genetic services.
RESUMO
A 70S ribosome was prepared from a 30S ribosome subunit from Euglena gracilis chloroplasts and a 50S ribosome subunit from Escherichia coli. This hybrid ribosome was active in polyuridylic acid-directed polyphenylalanine synthesis.
Assuntos
Cloroplastos , Escherichia coli , Hibridização Genética , Ribossomos , Fracionamento Celular , Centrifugação com Gradiente de Concentração , Euglena , Técnicas In Vitro , Fenilalanina/biossíntese , Polímeros/biossíntese , Ribossomos/análise , Nucleotídeos de Uracila/fisiologiaRESUMO
A new hepatotoxic microcystin was isolated from the cyanobacterium Microcystis aeruginosa strain 205. Its structure was found to be [Dha7]microcystin-RR as determined by amino acid analysis, mass spectrometry and 1H NMR spectroscopy. LD50 value (i.p. mouse) of this toxin was 180 micrograms/kg. The 48 hr lethal concentration (48-hr-LC50) of the toxin for larvae of the yellow fever mosquito, Aedes aegypti, was 14.9 micrograms/ml.
Assuntos
Microcystis/química , Peptídeos Cíclicos/química , Aedes , Sequência de Aminoácidos , Aminoácidos/análise , Animais , Doença Hepática Induzida por Substâncias e Drogas/fisiopatologia , Cromatografia Líquida de Alta Pressão , Larva , Dose Letal Mediana , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Microcistinas , Dados de Sequência Molecular , Peptídeos Cíclicos/toxicidadeRESUMO
Cultured cells of Anabaena flos-aquae strain CYA 83/1, isolated from Lake Edlandsvatn, Norway, produced two microcystin mono-methyl ester derivatives (1 and 2) at the D-Glu unit in addition to microcystin-LR (3), [D-Asp3]microcystin-LR (4), microcystin-RR (5), and [D-Asp3]microcystin-RR (6). Structures of these compounds were assigned based on their amino acid analysis with a Waters Pico Tag HPLC system plus fast atom bombardment mass spectrometry (FABMS), including tandem FABMS, analysis on the two new microcystins, [D-Glu(OCH3)6]microcystin-LR (1) and [D-Asp3, D-Glu(OCH3)6]microcystin-LR (2). Toxicity data were not obtained for 1 and 2 because of the small amounts isolated from the cells.
Assuntos
Anabaena/química , Peptídeos Cíclicos/análise , Sequência de Aminoácidos , Animais , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Cromatografia Líquida de Alta Pressão , Feminino , Toxinas Marinhas , Camundongos , Microcistinas , Dados de Sequência Molecular , Peptídeos Cíclicos/toxicidade , Espectrometria de Massas de Bombardeamento Rápido de ÁtomosRESUMO
Five microcystins were obtained from Microcystis aeruginosa strain CALU 972 isolated from a hepatotoxic water bloom collected in Lake Kroshnosero (Russia). The structure of a new toxin (1) was determined as [Dha7]microcystin-YR by amino acid analyses and fast atom bombardment mass spectrometry, and the toxins 2, 3, 4, and 5 were assigned the structures as [Dha7]microcystin-LR, [D-Asp3,Dha7]microcystin-LR, [Dha7]microcystin-RR, and [D-Asp3,Dha7]microcystin-RR, respectively, by direct comparison with authentic samples.
Assuntos
Microcystis/química , Peptídeos Cíclicos/isolamento & purificação , Sequência de Aminoácidos , Aminoácidos/análise , Toxinas Marinhas , Microcistinas , Dados de Sequência Molecular , Peptídeos Cíclicos/análise , Federação RussaRESUMO
A hepatotoxic cyanobacterial (blue-green algal) water bloom was collected from a constructed water reservoir in Finland. The water bloom contained two cyanobacterial species, Microcystis aeruginosa and Aphanizomenon flos-aquae. Two hepatotoxins, 1 and 2, were isolated from extracts of lyophilized cells. The structures of 1 and 2 were assigned based upon their amino acid analyses on a Waters Pico Tag HPLC system and a chiral GC capillary column (Chirasil Val III), fast atom bombardment mass spectrometry (FABMS), high resolution FABMS, and tandem FABMS data. Toxin 1 was identical to a previously reported compound, [D-Asp3]microcystin-RR. Toxin 2 was new and was assigned the structure [D-Asp3]microcystin-YR.
Assuntos
Cianobactérias/química , Peptídeos Cíclicos/isolamento & purificação , Aminoácidos/análise , Animais , Cromatografia Gasosa , Cromatografia Líquida de Alta Pressão , Hidrólise , Masculino , Toxinas Marinhas , Camundongos , Camundongos Endogâmicos ICR , Microcistinas , Peptídeos Cíclicos/análise , Espectrometria de Massas de Bombardeamento Rápido de ÁtomosRESUMO
Two new microcystins, [L-Ser7]microcystin-LR (1) and [L-Ser7]microcystin-RR (2), were isolated from a filamentous fresh water cyanobacterium (blue-green alga), Anabaena sp. strain 202 A1, along with the two major toxins, [Dha7]microcystin-LR (3) and [Dha7]microcystin-RR (4) and their minor components the D-Asp variants [D-Asp3,Dha7]microcystin-LR (5) and [D-Asp3,Dha7]microcystin-RR (6). Anabaena sp. strain 202 A1 also produced another new toxin, whose structure is tentatively proposed as [D-Asp3,L-Ser7]microcystin-XR (7), where X is a leucine homologue. Anabaena sp. strain 202 A2 produced one new microcystin, 1, and three known microcystins, 3, 4, and 5. The structures of the toxins were assigned based on their amino acid analyses, and fast atom bombardment mass spectrometry data.
Assuntos
Anabaena/química , Peptídeos Cíclicos/isolamento & purificação , Serina/análise , Sequência de Aminoácidos , Animais , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Cromatografia Gasosa , Feminino , Hidrólise , Toxinas Marinhas , Camundongos , Microcistinas , Dados de Sequência Molecular , Peso Molecular , Peptídeos Cíclicos/análise , Espectrometria de Massas de Bombardeamento Rápido de ÁtomosRESUMO
The reduction of acetylene to ethylene by stationary (non-shaking) cultures of free-living rhizobia under atmospheric oxygen levels has been demonstrated. Under these conditions the development of the activity is inhibited by 10 mM NH4Cl and about 20% of oxygen is required for maximal activity. When the stationary cultures were shaken, oxygen concentrations of 1% and higher were found to be inhibitory. Specific activities of 20 and 40 nmol of acetylene reduced h-1 mg-1 protein were observed.
Assuntos
Acetileno/metabolismo , Oxigênio , Rhizobium/metabolismo , Cloreto de Amônio/metabolismo , Pressão Atmosférica , Proteínas de Bactérias/biossíntese , Etilenos/metabolismo , OxirreduçãoRESUMO
The oxygen and nutritional requirements for acetylene reduction by Rhizobium japonicum and Rhizobium sp. in liquid culture are described. The optimal oxygen concentration was about 0.1% in the gas phase, which is lower than that of any other known aerobic nitrogen-fixing microorganism. these organisms are also unique in that nitrogenase synthesis is not repressed in the presence of ammonium chloride under certain cultural conditions, in contrast to other wild-type bacteria.
Assuntos
Acetileno/metabolismo , Consumo de Oxigênio , Rhizobium/metabolismo , Cloreto de Amônio/farmacologia , Repressão Enzimática , Nitrogenase/biossíntese , Oxirredução , Rhizobium/enzimologia , Rhizobium/crescimento & desenvolvimentoRESUMO
The water fern, Azolla caroliniana Willd., containing the symbiotic, heterocystous blue-green alga, Anabaena azollae, has been studied under various growth conditions to characterize its light-dependent production of H(2). The response of H(2) production to N(2) and C(2)H(2) and the absence of a differential effect of m-chlorocarbonyl cyanide phenylhydrazone on H(2) production and C(2)H(2) reduction, coupled with the parallel inhibition of both processes by DCMU imply that the production of H(2) is nitrogenase-catalyzed and ATP-dependent.H(2) was produced by fronds grown under air-CO(2) in the presence or absence of combined nitrogen. When cultured under argon-O(2)-CO(2), only those fronds provided with combined nitrogen remained viable and produced H(2). Fronds grown on nitrate under air plus 2% CO also produced H(2). In comparison to fronds grown on N(2) alone, fronds grown on nitrate had an increased rate of H(2) production relative to C(2)H(2) reduction, and the inhibition of H(2) production by air was less.CO in argon +/- CO(2) resulted in a partial inhibition of H(2) production, whereas CO in argon-CO(2)-C(2)H(2) enhanced H(2) production in fronds grown without combined nitrogen. Our studies strongly indicate that H(2) production is nitrogenase-catalyzed but the possibility that the symbiont contains a hydrogenase cannot be totally excluded.
RESUMO
Lyngbya wollei (Farlow ex Gomont) comb. nov., a perennial mat-forming filamentous cyanobacterium prevalent in lakes and reservoirs of the southeastern United States, was found to produce a potent, acutely lethal neurotoxin when tested in the mouse bioassay. Signs of poisoning were similar to those of paralytic shellfish poisoning. As part of the Tennessee Valley Authority master plan for Guntersville Reservoir, the mat-forming filamentous cyanobacterium L. wollei, a species that had recently invaded from other areas of the southern United States, was studied to determine if it could produce any of the known cyanotoxins. Of the 91 field samples collected at 10 locations at Guntersville Reservoir, Ala., on the Tennessee River, over a 3-year period, 72.5% were toxic. The minimum 100% lethal doses of the toxic samples ranged from 150 to 1,500 mg kg of lyophilized L. wollei cells-1, with the majority of samples being toxic at 500 mg kg-1. Samples bioassayed for paralytic shellfish toxins by the Association of Official Analytical Chemists method exhibited saxitoxin equivalents ranging from 0 to 58 micrograms g (dry weight)-1. Characteristics of the neurotoxic compound(s), such as the lack of adsorption by C18 solid-phase extraction columns, the short retention times on C18 high-performance liquid chromatography (HPLC) columns, the interaction of the neurotoxins with saxiphilin (a soluble saxitoxin-binding protein), and external blockage of voltage-sensitive sodium channels, led to our discovery that this neurotoxin(s) is related to the saxitoxins, the compounds responsible for paralytic shellfish poisonings. The major saxitoxin compounds thus far identified by comparison of HPLC fluorescence retention times are decarbamoyl gonyautoxins 2 and 3. There was no evidence of paralytic shellfish poison C toxins being produced by L. wollei. Fifty field samples were placed in unialgal culture and grown under defined culture conditions. Toxicity and signs of poisoning for these laboratory-grown strains of L. wollei were similar to those of the field collection samples.
Assuntos
Cianobactérias/metabolismo , Toxinas de Lyngbya/metabolismo , Toxinas de Lyngbya/toxicidade , Saxitoxina/metabolismo , Saxitoxina/toxicidade , Proteínas de Anfíbios , Animais , Bioensaio , Encéfalo/patologia , Proteínas de Transporte/metabolismo , Cromatografia Líquida de Alta Pressão , Cianobactérias/isolamento & purificação , Toxinas de Lyngbya/química , Masculino , Camundongos , Estrutura Molecular , Ratos , Saxitoxina/isolamento & purificação , Canais de Sódio/metabolismo , Sudeste dos Estados Unidos , Microbiologia da ÁguaRESUMO
Rhizobium strain BTAi 1, which nodulates both stems and roots of Aeschynomene indica L., formed bacteriochlorophyll and photosynthetic reaction centers resembling those of purple photosynthetic bacteria when grown aerobically ex planta under a light-dark cycle. Bacteriochlorophyll formation was not observed under continuous dark or light growth conditions. The amount of pigment formed was similar to that previously found in aerobic photosynthetic bacteria. Stem nodules appear to fix nitrogen photosynthetically, as illumination of A. indica stem nodules with near-infrared light resulted in an enhanced rate of acetylene reduction. Near-infrared light did not enhance acetylene reduction when either A. indica or soybean root nodules were illuminated. The BTAi 1 isolate can be differentiated from members of the family Rhodospirillaceae by several criteria.
RESUMO
A hepatotoxic strain of cyanobacterium Anabaena sp. 66 was isolated from a hepatotoxic water bloom sample in Lake Kiikkara, Finland. Four cyclic heptapeptide hepatotoxins were isolated and purified by HPLC from cultured cells of this organism. The structures of three new homotyrosine (Hty) containing toxins, [Dha7]microcystin-HtyR (Dha = dehydroalanine) (1), [D-Asp3,Dha7]microcystin-HtyR (2), and [L-Ser7]microcystin-HtyR (3), were assigned, based upon amino acid analyses using both a Waters Pico Tag HPLC system and chiral capillary GC, 1H NMR, fast atom bombardment mass spectrometry (FABMS), and collisionally induced tandem FABMS. A new homophenylalanine (Hph) variant of 1, [Dha7]microcystin-HphR (4), was also obtained as a minor component. Compound 3 is most likely a biosynthetic precursor of 1. The four new toxins did not have an N-methyl group at the dehydroamino acid or its precursor unit.
Assuntos
Anabaena/química , Toxinas Bacterianas/química , Peptídeos Cíclicos/química , Sequência de Aminoácidos , Aminoácidos/análise , Animais , Toxinas Bacterianas/isolamento & purificação , Toxinas Bacterianas/toxicidade , Cromatografia Líquida de Alta Pressão , Fígado/efeitos dos fármacos , Espectroscopia de Ressonância Magnética , Camundongos , Dados de Sequência Molecular , Peso Molecular , Peptídeos Cíclicos/isolamento & purificação , Peptídeos Cíclicos/toxicidade , Espectrometria de Massas de Bombardeamento Rápido de Átomos , Tirosina/análise , Valina/análise , Microbiologia da ÁguaRESUMO
Three new cyclic heptapeptide hepatotoxins, [D-Ser1,ADMAdda5]microcystin-LR (1), [D-Asp3,-ADMAdda5]microcystin-LHar (2), and [ADMAdda5,Mser7]microcystin-LR (3), were isolated from the cyanobacterium (blue-green alga) Nostoc sp. strain 152, together with four known microcystins, [ADMAdda5]microcystin-LR (4), [ADMAdda5]microcystin-LHar (5), [D-Asp3,-ADMAdda5]microcystin-LR (6), and [DMAdda5]microcystin-LR (7). The structures of new microcystins were assigned on the basis of high-resolution fast atom bombardment mass spectrometry (HR FABMS), collisionally induced tandem FABMS (FABMS/MS), amino acid analysis, and gas chromatography (GC) on a chiral capillary column. All three new toxins contained 9-acetoxy-3-amino-2,6,8-trimethyl-10-phenyldeca-4,6-dienoic acid (ADMAdda) instead of the corresponding 9-methoxyl derivative (Adda), while 7 contains the corresponding 9-hydroxy analog (DMAdda). Compound 1 is the first microcystin reported that contains D-serine (D-Ser) in lieu of the D-alanine (D-Ala) unit which was thought to be an invariable amino acid component of the microcystins. Compound 2 has L-homoarginine (Har) instead of L-arginine (L-Arg) in 6 and D-aspartic acid (D-Asp) instead of D-erythro-beta-methylaspartic acid (D-MeAsp) in 5. Compound 3, the N-methylserine (Mser) variant of the N-methyldehydroalanine unit in 4, would be a biosynthetic precursor of 4.
Assuntos
Toxinas Bacterianas/química , Cianobactérias/química , Fígado/efeitos dos fármacos , Peptídeos Cíclicos/química , Aminoácidos/análise , Toxinas Bacterianas/isolamento & purificação , Cromatografia Gasosa , Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Fina , Hidrólise , Microcistinas , Estrutura Molecular , Peso Molecular , Peptídeos Cíclicos/isolamento & purificação , Espectrometria de Massas de Bombardeamento Rápido de ÁtomosRESUMO
The infectivity of the soybean symbiont Rhizobium japonicum changed two- to fivefold with culture age for strains 110 ARS, 138 Str Spc, and 123 Spc, whereas culture age had relatively little effect on the infectivity of strains 83 Str and 61A76 Str. Infectivity was measured by determining the number of nodules which developed on soybean primary roots in the zone which contained developing and preemergent root hairs at the time of inoculation. Root cells in this region of the host root are susceptible to Rhizobium infection, but this susceptibility is lost during acropetal development and maturation of the root cells within a period of 4 to 6 h (T. V. Bhuvaneswari, B. G. Turgeon, and W. D. Bauer, Plant Physiol. 66:1027-1031, 1980). Profiles of nodulation frequency at different locations on the root were not affected by the age of the R. japonicum cultures, indicating that culture age affected the efficiency of Rhizobium infection rather than how soon infections were initiated after inoculation. Inoculum dose-response experiments also indicated that culture age affected the efficiency of infection. Two strains, 61A76 Str and 83 Str, were relatively inefficient at all culture ages, particularly at low inoculum doses. Changes in infectivity with culture age were reasonably well correlated with changes in the proportion of cells in a culture capable of binding soybean lectin. Suspensions of R. japonicum in water were found to retain their viability and infectivity.
Assuntos
Glycine max/microbiologia , Rhizobium/crescimento & desenvolvimento , Proteínas de Soja , Simbiose , Antibacterianos/farmacologia , Resistência Microbiana a Medicamentos , Lectinas , Mutação , Lectinas de Plantas , Rhizobium/metabolismo , Fatores de TempoRESUMO
Three Rhizobium japonicum strains and two slow-growing cowpea-type Rhizobium strains were found to remain viable and able to rapidly modulate their respective hosts after being stored in purified water at ambient temperatures for periods of 1 year and longer. Three fast-growing Rhizobium species did not remain viable under the same water storage conditions. After dilution of slow-growing Rhizobium strains with water to 10(3) to 10(5) cells ml-1, the bacteria multiplied until the viable cell count reached levels of between 10(6) and 10(7) cells ml-1. The viable cell count subsequently remained fairly constant. When the rhizobia were diluted to 10(7) cells ml-1, they did not multiply, but full viability was maintained. If the rhizobia were washed and suspended at 10(9) cells ml-1, viability slowly declined to 10(7) cells ml-1 during 9 months of storage. Scanning electron microscopy showed that no major morphological changes took place during storage. Preservation of slow-growing rhizobia in water suspensions could provide a simple and inexpensive alternative to current methods for the preservation of rhizobia for legume inoculation.
Assuntos
Técnicas Bacteriológicas , Fabaceae/microbiologia , Plantas Medicinais , Rhizobium/fisiologia , Microscopia Eletrônica de Varredura , Doenças das Plantas , Preservação Biológica/métodos , Rhizobium/citologia , Glycine max/microbiologia , Simbiose , Temperatura , Fatores de Tempo , Microbiologia da ÁguaRESUMO
Dihydro derivatives of nodularin (1) and microcystin-LR (4), potent cyclic peptide hepatotoxins isolated from Nodularia spumigena and Microcystis aeruginosa, respectively, were prepared by sodium borohydride reduction of the dehydroamino acid residues. The two stereoisomers of both dihydronodularin (2 and 3) and dihydromicrocystin-LR (5 and 6), isolated by reversed-phase HPLC, showed similar toxicity to each other [ip in mice, LD50 = 150 (2), 150 (3), 85 (5), and 100 (6) micrograms/kg]. The stereochemistries of the reduced amino acids obtained by acid hydrolysis of dihydronodularin and dihydromicrocystin-LR [respectively, alpha-(methylamino)butyric acid and N-methylalanine] were determined by GC on a permethylated beta-cyclodextrin capillary column as their trifluoroacetyl methyl ester derivatives. Authentic L- and DL-N-methylamino acids were prepared to compare directly with the natural amino acids. Deuterated derivatives were also prepared using sodium borodeuteride (98 atom % D), and the location (beta) and percentage (78-84%) of the deuterium incorporation were determined.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas/etiologia , Peptídeos Cíclicos/química , Peptídeos Cíclicos/toxicidade , Animais , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Cromatografia Gasosa , Cromatografia Líquida de Alta Pressão , Cianobactérias/imunologia , Cianobactérias/metabolismo , Toxinas de Cianobactérias , Hidrólise , Dose Letal Mediana , Espectroscopia de Ressonância Magnética , Masculino , Camundongos , Camundongos Endogâmicos ICR , Espectrometria de Massas de Bombardeamento Rápido de Átomos , Espectrofotometria Ultravioleta , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
Anabaena flos-aquae NRC 525-17 produces a very potent neurotoxin, anatoxin-a(s). During isolation of the neurotoxin, we found that the strain contains four other toxic compounds which show strong hepatotoxicity. The four toxins, toxins 1, 1', 2, and 3, were successfully purified. Toxin 2, one of major toxins, was identified as 3-desmethylmicrocystin LR (1) by comparison of spectral data of the known compound. Since the three other toxins contain an unknown amino acid, GC/MS was applied and it revealed the presence of homotyrosine in toxins 1 (2) and 1' (3). Only a partial structure was obtained for toxin 3 due to the small amount present in the cells.
Assuntos
Anabaena/metabolismo , Toxinas Bacterianas/análise , Aminoácidos/análise , Animais , Toxinas Bacterianas/isolamento & purificação , Toxinas Bacterianas/toxicidade , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Cromatografia Líquida de Alta Pressão , Liofilização , Cromatografia Gasosa-Espectrometria de Massas , Dose Letal Mediana , Espectroscopia de Ressonância Magnética , Masculino , Camundongos , Camundongos Endogâmicos ICR , Tamanho do Órgão/efeitos dos fármacosRESUMO
Hepatotoxins (microcystins) from seven freshwater Anabaena strains originating from three different Finnish lakes and one lake in Norway were isolated by high-performance liquid chromatography and characterized by amino acid analysis and fast atom bombardment mass spectrometry. All strains produced three to seven different microcystins. A total of 17 different compounds were isolated, of which 8 were known microcystins. The known compounds identified from six strains were MCYST (microcystin)-LR, [D-Asp3]MCYST-LR, [Dha7]MCYST-LR, [D-Asp3,Dha7]MCYST-LR, MCYST-RR, [D-Asp3]MCYST-RR, [Dha7]MCYST-RR, and [D-Asp3,Dha7]MCYST-RR. With the exception of MCYST-LR and [D-Asp3]MCYST-LR, this is the first time that isolation of these toxins from Anabaena strains has been reported. Three of the strains produced one to three toxins as minor components which could not be identified. Anabaena sp. strain 66 produced four unidentified toxins. The other Anabaena strains always contained both MCYST-LR and MCYST-RR and/or their demethyl variants. Quantitative differences between toxins within and between strains were detected; at times MCYST-LR and at other times MCYST-RR or demethyl derivatives thereof were the most abundant toxins found in a strain.