RESUMO
CASE: A 2.5-year-old male child presented to the clinic for evaluation of left wrist popping. Ten months earlier, he sustained a closed left both-bone forearm fracture (BBFF) treated with reduction and casting. His clinical course was complicated by redisplacement requiring secondary manipulation and casting before osseous union. His parents reported wrist popping with active motion in the setting of a 20° apex volar malunion of the midshaft radius. He has been treated with observation and monitoring of deformity remodeling. CONCLUSION: Distal radioulnar joint instability is a potential complication of malunited BBFF, even in a pediatric population. Residual deformity, especially in the radius, should prompt clinical follow-ups after osseous union to assess functional recovery and deformity remodeling.
Assuntos
Fraturas Mal-Unidas , Instabilidade Articular , Fraturas do Rádio , Criança , Pré-Escolar , Antebraço , Fraturas Mal-Unidas/complicações , Humanos , Instabilidade Articular/diagnóstico por imagem , Instabilidade Articular/etiologia , Instabilidade Articular/cirurgia , Masculino , Fraturas do Rádio/complicações , Fraturas do Rádio/diagnóstico por imagem , Fraturas do Rádio/cirurgia , Articulação do PunhoRESUMO
Understanding the mechanism(s) underpinning drug resistance could lead to novel treatments to reverse the increased tolerance of a pathogen. In this study, paromomycin (PMM) resistance (PMMr) was induced in three Nepalese clinical strains of Leishmania donovani with different inherent susceptibilities to antimony (Sb) drugs by stepwise exposure of promastigotes to PMM. Exposure to PMM resulted in the production of mixed populations of parasites, even though a single cloned population was used at the start of selection. PMM 50% inhibitory concentration (IC50) values for PMMr parasites varied between 104 and 481 µM at the promastigote stage and 32 and 195 µM at the intracellular amastigote stage. PMM resistance was associated with increased resistance to nitric oxide at the amastigote stage but not the promastigote stage (P < 0.05). This effect was most marked in the Sb-resistant (Sbr) PMMr clone, in which PMM resistance was associated with a significant upregulation of glutathione compared to that in its wild type (P < 0.05), although there was no change in the regulation of trypanothione (detected in its oxidized form). Interestingly, PMMr strains showed an increase in either the keto acid derivative of isoleucine (Sb intermediate PMMr) or the 2-hydroxy acids derived from arginine and tyrosine (Sb susceptible PMMr and Sbr PMMr). These results are consistent with the recent finding that the upregulation of the branched-chain amino acid aminotransferase and d-lactate dehydrogenase is linked to PMMr In addition, we found that PMMr is associated with a significant increase in aneuploidy during PMM selection in all the strains, which could allow the rapid selection of genetic changes that confer a survival advantage.
Assuntos
Antiprotozoários/farmacologia , Leishmania donovani/efeitos dos fármacos , Paromomicina/farmacologia , Animais , Resistência a Medicamentos/genética , Feminino , Genômica , Humanos , Leishmania donovani/genética , Leishmania donovani/metabolismo , Leishmaniose Visceral/tratamento farmacológico , Leishmaniose Visceral/parasitologia , Lipidômica , Macrófagos/parasitologia , Metabolômica , Camundongos , Camundongos Endogâmicos BALB C , Nepal , Testes de Sensibilidade Parasitária , Polimorfismo GenéticoRESUMO
ß-N-methylamino-L-alanine (BMAA) has been shown to accumulate in organisms by associating with host proteins. It has been proposed that this association is the result of misincorporation of BMAA into the primary structure of proteins, specifically in the place of L-serine, and that this misincorporation causes protein misfolding resulting in the tangle formation typically associated with neurodegenerative diseases. However, more recent studies have questioned the validity of the BMAA misincorporation hypothesis. Furthermore, BMAA association with proteins in the absence of de novo protein synthesis has been demonstrated although the nature of these associations has not yet been characterized. We therefore sought to investigate the effects of these undescribed interactions on protein functioning, and to identify the site(s) of these interactions. We present data here to show that BMAA can inhibit the activity of certain enzymes, interfere with protein folding in the absence of de novo protein synthesis, and associate in vitro with commercial proteins to such an extent that it cannot be removed by protein precipitation or denaturation. Based on the observed effects of these interactions on protein functioning, we suggest that this might constitute an additional mechanism of toxicity that could help to explain the role of BMAA in the development of neurodegenerative diseases.
Assuntos
Diamino Aminoácidos/química , Diamino Aminoácidos/toxicidade , Enzimas/química , Dobramento de Proteína/efeitos dos fármacos , Caseínas/química , Toxinas de Cianobactérias , Melaninas/química , Neurotoxinas/química , Ribonuclease H/químicaRESUMO
The implication of ß-N-methylamino-L-alanine (BMAA) in the development of neurodegenerative diseases worldwide has led to several investigations of the mechanism, or mechanisms, of toxicity of this cyanobacterially produced amino acid. The primary mechanism of toxicity that was identified is excitotoxicity, with a second possible mechanism, the misincorporation of BMAA into the primary protein structure and consequent cell damage, having been more recently reported. However, studies on excitotoxicity and misincorporation have been conducted independently and there are therefore no data available on the relative contribution of each of these mechanisms to the total toxicity of BMAA. The rat pheochromocytoma cell line PC12 is an ideal model for a study of this type, as glutamate receptor expression is modified by cell differentiation, which can be affected by exposure to nerve growth factor. In this study, the PC12 cell line was evaluated as a model to study BMAA toxicity via the two proposed mechanisms: excitotoxicity and protein misincorporation. BMAA and canavanine treatment of cultures of PC12 were evaluated for depolarization of the mitochondrial membrane. In canavanine-treated cultures, this was evident after 9 days of treatment and was attributed to the primary mechanism of canavanine toxicity, protein misincorporation. However, no membrane depolarization was observed for BMAA-treated cultures even after 21 days of continuous treatment at 500 µM. Short-term exposure to both BMAA and canavanine resulted in a slight increase in necrosis in undifferentiated cells that was prevented in canavanine-treated cultures by co-incubation with arginine, but not in BMAA-treated cultures by co-incubation with serine. A slight increase in apoptosis was observed in undifferentiated cells treated with either BMAA or glutamate, and ROS production increased in glutamate-treated cells. However, the excitotoxicity was less pronounced than reported in previous studies with neuronal cells. In contrast, apoptosis was greatly increased in both BMAA- and glutamate-treated cells after differentiation and resulting mGluR1 increase, indicating that excitotoxicity is the main, if not only, mechanism of toxicity in PC12.
Assuntos
Diamino Aminoácidos/toxicidade , Agonistas de Aminoácidos Excitatórios/toxicidade , Neurônios/efeitos dos fármacos , Diamino Aminoácidos/análise , Animais , Anexina A5/metabolismo , Apoptose/efeitos dos fármacos , Arginina/metabolismo , Transporte Biológico/efeitos dos fármacos , Canavanina/análise , Canavanina/farmacologia , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Toxinas de Cianobactérias , Ácido Glutâmico/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Fator de Crescimento Neural/farmacologia , Células PC12/efeitos dos fármacos , Fosfatidilserinas/metabolismo , Ratos , Espécies Reativas de Oxigênio/metabolismo , Fatores de TempoRESUMO
In this study, we followed the genomic, lipidomic and metabolomic changes associated with the selection of miltefosine (MIL) resistance in two clinically derived Leishmania donovani strains with different inherent resistance to antimonial drugs (antimony sensitive strain Sb-S; and antimony resistant Sb-R). MIL-R was easily induced in both strains using the promastigote-stage, but a significant increase in MIL-R in the intracellular amastigote compared to the corresponding wild-type did not occur until promastigotes had adapted to 12.2 µM MIL. A variety of common and strain-specific genetic changes were discovered in MIL-adapted parasites, including deletions at the LdMT transporter gene, single-base mutations and changes in somy. The most obvious lipid changes in MIL-R promastigotes occurred to phosphatidylcholines and lysophosphatidylcholines and results indicate that the Kennedy pathway is involved in MIL resistance. The inherent Sb resistance of the parasite had an impact on the changes that occurred in MIL-R parasites, with more genetic changes occurring in Sb-R compared with Sb-S parasites. Initial interpretation of the changes identified in this study does not support synergies with Sb-R in the mechanisms of MIL resistance, though this requires an enhanced understanding of the parasite's biochemical pathways and how they are genetically regulated to be verified fully.
Assuntos
Antiprotozoários/farmacologia , Leishmania donovani/efeitos dos fármacos , Leishmania donovani/metabolismo , Fosforilcolina/análogos & derivados , Animais , Antimônio/farmacologia , Resistência a Medicamentos , Feminino , Leishmania donovani/genética , Leishmaniose Visceral/parasitologia , Metabolismo dos Lipídeos , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Mutação , Nepal , Testes de Sensibilidade Parasitária , Fosforilcolina/farmacologiaRESUMO
The most significant modulators of the cyanotoxins microcystin and ß-N-methylamino-L-alanine in laboratory cyanobacterial cultures are the concentration of growth-medium combined nitrogen and nitrogen uptake rate. The lack of field studies that support these observations led us to investigate the cellular content of these cyanotoxins in cyanobacterial bloom material isolated from a freshwater impoundment and to compare these to the combined nitrogen availability. We established that these toxins typically occur in an inverse relationship in nature and that their presence is mainly dependent on the environmental combined nitrogen concentration, with cellular microcystin present at exogenous combined nitrogen concentrations of 29 µM and higher and cellular BMAA correlating negatively with exogenous nitrogen at concentrations below 40 µM. Furthermore, opposing nutrient and light gradients that form in dense cyanobacterial blooms may result in both microcystin and BMAA being present at a single sampling site.
Assuntos
Diamino Aminoácidos/metabolismo , Carcinógenos/metabolismo , Meio Ambiente , Microcistinas/metabolismo , Microcystis/metabolismo , Nitrogênio/farmacologia , Amônia/farmacologia , Clorofila/metabolismo , Clorofila A , Toxinas de Cianobactérias , Água Doce/análise , Microcystis/efeitos dos fármacos , Nitratos/farmacologia , Nitritos/farmacologiaRESUMO
ß-N-Methylamino-L-alanine, an unusual amino acid implicated in neurodegenerative disease, has been detected in cultures of nearly all genera of environmentally ubiquitous cyanobacteria tested. The compound is present within cyanobacterial cells in free and protein-associated forms, with large variations occurring in the concentration of these pools between species as well as within single strains. With a lack of knowledge and supporting data on the regulation of BMAA production and the role of this compound in cyanobacteria, the association between BMAA and cyanobacteria is still subject to debate. In this study we investigated the biosynthesis of BMAA in axenic non-diazotrophic cyanobacterial cultures using the stable isotope ¹5N. Nitrogen starvation of nutritionally replete cells resulted in an increase in free cellular ¹5N BMAA suggesting that BMAA may be the result of catabolism to provide nitrogen or that BMAA is synthesised to serve a functional role in the cell in response to nitrogen deprivation. The addition of NO3â» and NH4⺠to the culture medium following starvation resulted in a decrease of free cellular BMAA without a corresponding increase in the protein-associated fraction. The use of ammonia as a nitrogen source resulted in a more rapid reduction of BMAA when compared to nitrate. This study provides the first data regarding the regulation of intracellular BMAA concentrations in cyanobacteria with results conclusively showing the production of ¹5N BMAA by an axenic cyanobacterial culture.
Assuntos
Diamino Aminoácidos/biossíntese , Toxinas Bacterianas/biossíntese , Cianobactérias/metabolismo , Neurotoxinas/biossíntese , Nitrogênio/deficiência , Diamino Aminoácidos/química , Diamino Aminoácidos/metabolismo , Amônia/metabolismo , Cultura Axênica , Toxinas Bacterianas/química , Toxinas Bacterianas/metabolismo , Cromatografia Líquida de Alta Pressão , Toxinas de Cianobactérias , Toxinas Marinhas/biossíntese , Toxinas Marinhas/química , Toxinas Marinhas/metabolismo , Microcystis/metabolismo , Neurotoxinas/química , Neurotoxinas/metabolismo , Nitratos/metabolismo , Isótopos de Nitrogênio , Especificidade da Espécie , Espectrometria de Massas por Ionização por Electrospray , Estatística como Assunto , Synechocystis/metabolismo , Espectrometria de Massas em Tandem , Fatores de TempoRESUMO
ß-N-methylamino-L-alanine (BMAA) is produced by diverse taxa of cyanobacteria, and has been detected by many investigators who have searched for it in cyanobacterial blooms, cultures and collections. Although BMAA is distinguishable from proteinogenic amino acids and its isomer 2,4-DAB using standard chromatographic and mass spectroscopy techniques routinely used for the analysis of amino acids, we studied whether BMAA could be reliably distinguished from other diamino acids, particularly 2,6-diaminopimelic acid which has been isolated from the cell walls of many bacterial species. We used HPLC-FD, UHPLC-UV, UHPLC-MS, and triple quadrupole tandem mass spectrometry (UHPLC-MS/MS) to differentiate BMAA from the diamino acids 2,6-diaminopimelic acid, N-2(amino)ethylglycine, lysine, ornithine, 2,4-diaminosuccinic acid, homocystine, cystine, tryptophan, as well as other amino acids including asparagine, glutamine, and methionine methylsulfonium.
Assuntos
Diamino Aminoácidos/química , Aminoácidos Dicarboxílicos/química , Cianobactérias/química , Cromatografia Líquida de Alta Pressão/métodos , Ácido Diaminopimélico/química , Espectrometria de MassasRESUMO
Cyanobacteria are known to produce bioactive secondary metabolites such as hepatotoxins, cytotoxins and neurotoxins. The newly recognized neurotoxin ß-N-methylamino-L-alanine (BMAA) is a naturally occurring non-protein amino acid found in the majority of cyanobacterial genera tested. Evidence that exists for implication of BMAA in neurodegenerative disorders relies on bioaccumulation and biomagnification from symbiotic cyanobacteria. Uptake and accumulation of free BMAA by various non-symbiotic organisms, including aquatic macrophytes, has been documented but to date limited evidence of ecotoxicology exists. We therefore investigated the effect of BMAA on the oxidative stress responses of the macrophyte, Ceratophyllum demersum. Markers for oxidative stress in this study are the antioxidative enzymes superoxide dismutase, catalase, guaiacol peroxidase, glutathione peroxidase and glutathione reductase. We found that BMAA had an inhibitory effect on all the oxidative stress response enzymes tested in plants exposed to BMAA. However enzymes not related to oxidative stress response were not affected by BMAA in in vitro experiments. Binding studies in the presence of BMAA showed reduced enzyme specific activity over time compared to the control. This study shows that BMAA causes oxidative stress indirectly as it inhibits antioxidant enzymes required to combat reactive oxygen species that cause damage to cells. Further investigations are required to fully understand the inhibitory effect of BMAA on these enzymes.
Assuntos
Diamino Aminoácidos/metabolismo , Diamino Aminoácidos/toxicidade , Toxinas Bacterianas/toxicidade , Cianobactérias/química , Magnoliopsida/enzimologia , Estresse Oxidativo/efeitos dos fármacos , Análise de Variância , Catalase/antagonistas & inibidores , Toxinas de Cianobactérias , Glutationa Peroxidase/antagonistas & inibidores , Glutationa Redutase/antagonistas & inibidores , Cinética , Magnoliopsida/efeitos dos fármacos , Peroxidase/antagonistas & inibidores , Ligação Proteica , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/antagonistas & inibidoresRESUMO
Free-living freshwater cyanobacteria contain BMAA in both free cellular and protein-associated forms. Free BMAA released on bloom collapse or during cellular turnover creates a potential source of the non-proteinogenic amino acid for bioaccumulation and biomagnification in aquatic ecosystems. Uptake of free amino acids is well documented in macrophytes and the potential for aquatic macrophytes to bioaccumulate BMAA therefore poses a potential threat where such macrophytes constitute a food source in an ecosystem. BMAA uptake and accumulation by the aquatic macrophyte Ceratophyllum demersum was therefore investigated. Rapid uptake of significant amounts of BMAA was observed in C. demersum. Both free and protein-associated BMAA were observed with protein association following accumulation of free BMAA. The protein association suggests potential biomaccumulation by aquatic macrophytes and offers a possibility of phytoremediation for BMAA removal.
Assuntos
Diamino Aminoácidos/toxicidade , Ecossistema , Magnoliopsida/efeitos dos fármacos , Neurotoxinas/toxicidade , Poluentes Químicos da Água/toxicidade , Diamino Aminoácidos/metabolismo , Cianobactérias/química , Cianobactérias/metabolismo , Toxinas de Cianobactérias , Água Doce , Magnoliopsida/crescimento & desenvolvimento , Magnoliopsida/metabolismo , Neurotoxinas/metabolismo , Poluentes Químicos da Água/metabolismoRESUMO
The cyanobacterial neurotoxin beta-N-methylamino-L-alanine (BMAA) has been associated with certain forms of progressive neurodegenerative disease, including sporadic Amyotrophic Lateral Sclerosis and Alzheimer's disease. Reports of BMAA in cyanobacterial blooms from lakes, reservoirs, and other water resources continue to be made by investigators in a variety of laboratories. Recently it was suggested that during analysis BMAA may be confused with its structural isomer 2,4-diaminobutyric acid (2,4-DAB), or that current detection methods may mistake other compounds for BMAA. We here review the evidence that BMAA can be consistently and reliably separated from 2,4-DAB during reversed-phase HPLC, and that BMAA can be confidently distinguished from 2,4-DAB during triple quadrupole LC-MS/MS analysis by i) different retention times, ii) diagnostic product ions resulting from collision-induced dissociation, and iii) consistent ratios between selected reaction monitoring (SRM) transitions. Furthermore, underivatized BMAA can be separated from 2,4-DAB with an amino acid analyzer with post-column visualization using ninhydrin. Other compounds that may be theoretically confused with BMAA during chloroformate derivatization during GC analysis are distinguished due to their different retention times.
Assuntos
Diamino Aminoácidos/análise , Aminobutiratos/análise , Toxinas Bacterianas/análise , Cianobactérias/química , Neurotoxinas/análise , Cromatografia Líquida de Alta Pressão , Toxinas de Cianobactérias , Isomerismo , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em TandemRESUMO
The chicken lysozyme gene encodes a hydrolase that has a key role in defence, especially in ovo. This gene was resequenced in global chicken populations [red, grey, Ceylon and green jungle fowl (JF)] and related bird species. Networks, summary statistics and tests of neutrality indicate that although there is extensive variation at the gene, little is present at coding sites, with the exception of one non-synonymous site. This segregating site and a further fixed non-synonymous change between red JF and domestic chicken populations are spatially close to the catalytic sites of the enzyme and so might affect its activity.
Assuntos
Galinhas/genética , Muramidase/genética , Animais , Galinhas/metabolismo , Genética Populacional , Modelos Moleculares , Muramidase/química , Filogenia , Polimorfismo de Nucleotídeo ÚnicoRESUMO
BACKGROUND: Chronic aspiration associated with gastro oesophageal reflux disease (GERD) is thought to play a substantial role in the development of asthma, the incidence of which is dramatically increasing in industrially developed countries. The majority of data examining the association between aspiration and asthma has been obtained from epidemiological studies, which show that between 50 and 90% of individuals with asthma experience some element of GERD. This study describes the effect of chronic aspiration on a model of experimentally induced airway hypersensitivity in Balb/c mice. MATERIALS AND METHODS: Four experimental groups were utilized: Aspiration/Asthma, Sham/Asthma, Aspiration/Sham and Sham/Sham. Mice were sensitized with aerosolized 1% ovalbumin on days 1 to 10 (sensitization phase), followed by repeated exposure on days 31 to 40 (challenge phase). Aspiration events occurred on days 1, 8,15, 22, 29, 36, 43 and 50. Animals were sacrificed on days 56 and 57. RESULTS: Chronic aspiration of 10 microL of murine gastric fluid per week for eight weeks produced an injury pattern distinct from that of acute aspiration, with lung injury characterized by hyperplasia, neutrophil infiltration of the bronchioles and relative parenchymal sparing. Aspiration during induction of ovalbumin-induced airway hypersensitivity was associated with a trend toward decreased production of antiovalbumin IgG, antiovalbumin IgE, and total IgE. Further, aspiration induced a substantial and significant increase in antiovalbumin IgG1/IgG2a ratios, consistent with a shift toward a predominantly Th2 response. CONCLUSION: These findings indicate that chronic aspiration has a profound effect on the nature of the immune response to aerosolized allergens in a model of experimentally induced airway hypersensitivity.
Assuntos
Asma/imunologia , Hiper-Reatividade Brônquica/imunologia , Líquido da Lavagem Broncoalveolar/química , Citocinas/análise , Imunoglobulina E/imunologia , Animais , Broncoconstritores/administração & dosagem , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Feminino , Pulmão/patologia , Camundongos , Ovalbumina/administração & dosagemRESUMO
Beta-N-methylamino-L-alanine (BMAA) is a neurotoxic non-proteinogenic amino acid reportedly produced by the majority of cyanobacterial isolates. A novel method was developed for the detection of BMAA in biological samples. Cultures representing the taxonomic diversity and geographic distribution in Southern Africa were collected and made uni-algal by standard methods before analysis for the presence of both free and protein-associated BMAA. Protein-associated BMAA was released by acid hydrolysis in an inert atmosphere. Samples were analyzed by gas chromatography-mass spectrometry (GC-MS) with pre-derivatization of amino acids using Phenomonex EZ:faast of the tested cultures, 96% were positive for BMAA although several were below the limit for quantification. BMAA presence was not related to the geographic origin or taxonomy of isolates and no correlation between free and bound BMAA concentrations was observed within or between taxonomic groups. These data offer the first confirmation of the taxonomic and geographic ubiquity of BMAA in freshwater cyanobacteria.
Assuntos
Diamino Aminoácidos/análise , Cianobactérias/química , Diamino Aminoácidos/metabolismo , Cianobactérias/metabolismo , Toxinas de Cianobactérias , Cromatografia Gasosa-Espectrometria de Massas , Sensibilidade e Especificidade , África do SulRESUMO
Hepatotoxin production in cyanobacteria has been shown to correlate to external stimuli such as light and nutrient concentrations and ratios, although conflicting results have been reported. Specific growth rates and protein and microcystin content of M. aeruginosa PCC7806 and M. aeruginosa UV027 were determined under nonlimiting batch culture conditions for a range of medium nitrogen and phosphorous atomic ratios. Both strains exhibited a similar optimal medium N:P ratio for increased cellular microcystin levels. Additionally, total cellular protein content and intracellular microcystin content were significantly correlated to each other (r2 = 0.81, p < 0.001). Microcystin and protein content increased considerably as the maximum specific growth rate for the experimental conditions was reached. The significant correlation of cellular protein and microcystin content and their relative increase with increasing specific growth rate, within defined ranges of medium N:P ratios, suggest a close association between microcystin production and N:P ratio-dependent assimilation of nitrogen, and resulting total cellular protein levels, which may be further modulated by specific growth rate.
Assuntos
Carcinógenos/análise , Microcystis/química , Microcystis/crescimento & desenvolvimento , Nitrogênio/metabolismo , Peptídeos Cíclicos/análise , Fósforo/metabolismo , Microcistinas , Nitrogênio/análise , Fósforo/análise , Dinâmica Populacional , Proteínas/análiseRESUMO
Modulation of microcystin production has been extensively studied in both batch and continuous cultures. Positive correlations with medium nitrogen, medium phosphorous, light intensity, inorganic carbon availability, and growth rate have been reported. Negative correlations have been reported between microcystin content and medium phosphorous. The only reported quantitative relationship between any variable and microcystin production was that of growth rate. Microcystis aeruginosa PCC7806 was therefore cultured under continuous culture conditions in a bubble-lift reactor at a growth rate of 0.01 h(-1) in modified BG11 (constant phosphate concentration of 0.195 mM and varying nitrate from 0.125 to 18 mM) and sampled at steady states for analysis of cell number, microcystin content, cellular N and P, residual medium nutrient concentration, and carbon fixation rate. Cellular microcystin quotas showed significant positive correlation with both nitrate uptake and cellular nitrogen content and were negatively correlated with carbon fixation rate, phosphate uptake, and cellular phosphorous. Thus, the ratio of nitrate uptake to phosphate uptake, cellular N to cellular P, and nitrate uptake to carbon fixation were positively correlated to cellular microcystin. Microcystin quotas increased 10-fold from the lowest to the highest steady-state values. Cellular microcystin content therefore is controlled to a significant extent by variables other than growth rate, as was previously reported, with nitrogen the most significant modulator. Batch culture in BG11 under identical conditions yielded increased microcystin when nitrogen uptake exceeded relative growth rate, confirming the importance of nitrogen uptake in the modulation of microcystin content for a specific growth rate.
Assuntos
Carbono/metabolismo , Microcystis/química , Nitrogênio/farmacocinética , Peptídeos Cíclicos/análise , Peptídeos Cíclicos/biossíntese , Inibidores Enzimáticos/análise , Microcistinas , Microcystis/crescimento & desenvolvimento , Microcystis/fisiologia , Nitratos/farmacocinética , Fosfatos/farmacocinéticaRESUMO
This paper introduces broad concepts of vulnerability, food security and famine. It argues that the concepts and theories driving development and implementation of vulnerability assessment tools are related to their utility. The review concludes that socio-geographic scale is a key issue, and challenge. It analyses three vulnerability assessment (VA) methods, using Ethiopia as a case study. Facing the challenges of vulnerability assessment and early warning requires providing accurate information at the required scale, useful for multiple decision-makers within realistic institutional capacities.
Assuntos
Abastecimento de Alimentos , Medição de Risco/métodos , Inanição/prevenção & controle , Algoritmos , Etiópia , Humanos , Modelos Estatísticos , Redes Neurais de Computação , Fatores SocioeconômicosRESUMO
A method is described for the quantification of mitochondrial DNA present in crude biological preparations. A known copy number of a standard is amplified in the presence of inactivated target DNA so as to determine the overall efficiency of the PCR process in a particular sample. In this way any inhibitory and/or stimulatory substances present in sample preparations can be taken into account. To reduce tube-to-tube variations product DNA quantification is limited to small cycle numbers. Using this method quantitations of DNA amounts in different crude preparations can be compared.
Assuntos
DNA Mitocondrial/análise , Reação em Cadeia da Polimerase/métodos , Animais , SuínosRESUMO
The DNA sequence and preliminary functional analysis of a 103-kbp section of the left arm of yeast chromosome I is presented. This region, from the left telomere to the LTE1 gene, can be divided into two distinct portions. One portion, the telomeric 29 kbp, has a very low gene density (only five potential genes and 21 kbp of noncoding sequence), does not encode any "functionally important" genes, and is rich in sequences repeated several times within the yeast genome. The other portion, with 37 genes and only 14.5 kbp of noncoding sequence, is gene rich and codes for at least 16 "functionally important" genes. The entire gene-rich portion is apparently duplicated on chromosome XV as an extensive region of partial gene synteney called a cluster homology region. A function can be assigned with varying degrees of precision to 23 of the 42 potential genes in this region; however, the precise function is know for only eight genes. Nineteen genes encode products presently novel to yeast, although five of these have homologs elsewhere in the yeast genome.
Assuntos
Cromossomos Fúngicos , Genes Fúngicos , Família Multigênica , Saccharomyces cerevisiae/genética , Sequência de Bases , DNA Fúngico , Dados de Sequência Molecular , Fases de Leitura Aberta , Homologia de Sequência do Ácido NucleicoRESUMO
The UFE1 gene of Saccharomyces cerevisiae was cloned, sequenced and characterized. The coding region of UFE1 is separated from the TMP1 gene on chromosome XV by 624 bp. Gene-disruption experiments demonstrated that UFE1 is essential for both the germination of ascospores and for vegetative growth. Translation of the UFE1 coding region generates a protein with significant similarity to cytokeratin and to the coiled-coil region of SED5, USO1 and restin, suggesting that it is involved in the secretory pathway and may also be related to intermediate filament-associated proteins.