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1.
Microorganisms ; 8(2)2020 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-32098253

RESUMO

Several artisanal cheeses are elaborated in European countries, being commonly curdled with rennets of animal origin. However, in some Spanish regions some cheeses of type "Torta" are elaborated using Cynara cardunculus L. rennets. Two of these cheeses, "Torta del Casar" and "Torta de Trujillo", are elaborated in Cáceres province with ewe's raw milk and matured over at least 60 days without starters. In this work, we identified the lactic acid bacteria present in these cheeses using MALDI-TOF MS and pheS gene analyses, which showed they belong to the species Lactobacillus curvatus, Lactobacillus diolivorans, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactococcus lactis and Leuconostoc mesenteroides. The pheS gene analysis also allowed the identification of the subspecies La. plantarum subsp. plantarum, La. paracasei subsp. paracasei and Le. mesenteroides subsp. jonggajibkimchii. Low similarity values were found in this gene for some currently accepted subspecies of Lc. lactis and for the two subspecies of La. plantarum, and values near to 100% for the subspecies of Le. mesenteroides and La. paracasei. These results, which were confirmed by the calculated ANIb and dDDH values of their whole genomes, showed the need to revise the taxonomic status of these species and their subspecies.

2.
Syst Appl Microbiol ; 42(6): 126019, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31635886

RESUMO

Phaseolus vulgaris is a legume indigenous to America which is currently cultivated in Europe including countries located at the Southeast of this continent, such as Croatia, where several local landraces are cultivated, most of them of Andean origin. In this work we identify at species and symbiovar levels several fast-growing strains able to form effective symbiosis with P. vulgaris in different Croatian soils. The identification at species level based on MALDI-TOF MS and core gene sequence analysis showed that most of these strains belong to the species R. leguminosarum, R. hidalgonense and R. pisi. In addition, several strains belong to putative new species phylogenetically close to R. ecuadorense and R. sophoriradicis. All Croatian strains belong to the symbiovar phaseoli and harbour the α and γ nodC alleles typical for American strains of this symbiovar. Nevertheless, most of Croatian strains harboured the γ nodC gene allele supporting its Andean origin since it is also dominant in other European countries, where Andean cultivars of P. vulgaris are traditionally cultivated, as occurs in Spain. The only strains harbouring the α nodC allele belong to R. hidalgonense and R. pisi, this last only containing the symbiovars viciae and trifolii to date. This is the first report about the presence in Europe of the species R. hidalgonense, the nodulation of P. vulgaris by R. pisi and the existence of the symbiovar phaseoli within this species. These results significantly increase the knowledge of the biogeography of Rhizobium-P. vulgaris symbiosis.


Assuntos
Biodiversidade , Phaseolus/microbiologia , Filogenia , Rhizobium/classificação , Rhizobium/genética , Nódulos Radiculares de Plantas/microbiologia , Proteínas de Bactérias/genética , Croácia , DNA Bacteriano/genética , Técnica de Amplificação ao Acaso de DNA Polimórfico , Rhizobium/química , Análise de Sequência de DNA , Microbiologia do Solo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Simbiose/genética
3.
Adv Exp Med Biol ; 1073: 125-135, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31236841

RESUMO

Urine is a biological fluid that can be collected noninvasively in relatively large quantities which can be used for the search of biomarkers of disease, both diseases of the urological tract and systemic diseases. One of the most important aspects in proteomic studies is sample treatment before further analysis. Methods of preparation of a urine sample depend on the techniques that will be used later for separation and identification of the proteins. Also, urine preparation should be as simple as possible to increase reproducibility. Normal urine has a much diluted protein concentration with a high-salt content, which interferes with proteomic analysis. Thus, an initial step in the handling of urine sample should be to concentrate and eliminate salts. As range of protein concentrations in urine spans several orders of magnitude, effective proteomic analyses require either removal of most abundant protein or enrichment of the less abundant ones. In this chapter, we discuss the aspects related to the collection and treatment of urine for proteomic studies.


Assuntos
Proteômica , Manejo de Espécimes/métodos , Urinálise/métodos , Biomarcadores/urina , Humanos , Proteínas/análise , Reprodutibilidade dos Testes
4.
Syst Appl Microbiol ; 42(2): 240-247, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30415881

RESUMO

Phaseolus vulgaris and Lens culinaris are two legumes with different distribution centers that were introduced in Spain at different times, but in some regions L. culinaris has been traditionally cultivated and P. vulgaris did not. Here we analysed the rhizobia isolated from nodules of these two legumes in one of these regions. MALDI-TOF MS analysis showed that all isolated strains matched with Rhizobium laguerreae and the phylogenetic analysis of rrs, atpD and recA genes confirmed these results. The phylogenetic analysis of these core genes allowed the differentiation of several groups within R. laguerreae and unexpectedly, strains with housekeeping genes identical to that of the type strain of R. laguerreae presented some differences in the rrs gene. In some strains this gene contains an intervening sequence (IVS) identical to that found in Rhizobium strains nodulating several legumes in different geographical locations. The atpD, recA and nodC genes of all isolated strains clustered with those of strains nodulating L. culinaris in its distribution centers, but not with those nodulating P. vulgaris in theirs. Therefore, all these strains belong to the symbiovar viciae, including those isolated from P. vulgaris, which in the studied region established effective symbiosis with the common endosymbiont of L. culinaris, instead to with its common endosymbiont, the symbiovar phaseoli. These results are particularly interesting for biogeography studies, because they showed that, due its high promiscuity degree, P. vulgaris is able to establish symbiosis with local symbiovars well established in the soil after centuries of cultivation with other legumes.


Assuntos
Phaseolus/microbiologia , Filogenia , Rhizobium/classificação , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , Genes Bacterianos , Rhizobium/isolamento & purificação , Nódulos Radiculares de Plantas/microbiologia , Análise de Sequência de DNA , Microbiologia do Solo , Espanha
5.
Enferm. infecc. microbiol. clín. (Ed. impr.) ; 35(5): 303-313, mayo 2017. graf
Artigo em Espanhol | IBECS | ID: ibc-162762

RESUMO

La espectrometría de masas MALDI-TOF es ya una herramienta de trabajo rutinaria en Microbiología Clínica, por su rapidez y fiabilidad en la identificación de microorganismos. Sus resultados están perfectamente contrastados en la identificación de bacterias y levaduras. La identificación de micobacterias y hongos filamentosos presenta mayor complejidad, por la mayor heterogeneidad de espectros dentro de cada especie. La metodología es algo más compleja, y la ampliación del número de especies de referencia, y del número de espectros de cada especie, serán cruciales para alcanzar mayor eficacia. La identificación directa a partir de hemocultivos se ha implantado dada su aportación al manejo de pacientes graves, pero su aplicación a otras muestras es más compleja. Los medios de cultivos cromogénicos han supuesto también una aportación al diagnóstico rápido tanto en bacterias como en levaduras, ya que aceleran el diagnóstico, facilitan la detección de cultivos mixtos y permiten un diagnóstico rápido de especies resistentes


MALDI-TOF mass spectrometry is now a routine resource in Clinical Microbiology, because of its speed and reliability in the identification of microorganisms. Its performance in the identification of bacteria and yeasts is perfectly contrasted. The identification of mycobacteria and moulds is more complex, due to the heterogeneity of spectra within each species. The methodology is somewhat more complex, and expanding the size of species libraries, and the number of spectra of each species, will be crucial to achieve greater efficiency. Direct identification from blood cultures has been implemented, since its contribution to the management of severe patients is evident, but its application to other samples is more complex. Chromogenic media have also contributed to the rapid diagnosis in both bacteria and yeast, since they accelerate the diagnosis, facilitate the detection of mixed cultures and allow rapid diagnosis of resistant species


Assuntos
Humanos , Infecções Bacterianas/microbiologia , Micoses/microbiologia , Bactérias/isolamento & purificação , Fungos/isolamento & purificação , Espectrometria de Massas , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
6.
Enferm Infecc Microbiol Clin ; 35(5): 303-313, 2017 May.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-28108122

RESUMO

MALDI-TOF mass spectrometry is now a routine resource in Clinical Microbiology, because of its speed and reliability in the identification of microorganisms. Its performance in the identification of bacteria and yeasts is perfectly contrasted. The identification of mycobacteria and moulds is more complex, due to the heterogeneity of spectra within each species. The methodology is somewhat more complex, and expanding the size of species libraries, and the number of spectra of each species, will be crucial to achieve greater efficiency. Direct identification from blood cultures has been implemented, since its contribution to the management of severe patients is evident, but its application to other samples is more complex. Chromogenic media have also contributed to the rapid diagnosis in both bacteria and yeast, since they accelerate the diagnosis, facilitate the detection of mixed cultures and allow rapid diagnosis of resistant species.


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Técnicas de Tipagem Micológica/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Bactérias/classificação , Bactérias/isolamento & purificação , Infecções Bacterianas/microbiologia , Líquidos Corporais/microbiologia , Compostos Cromogênicos , Meios de Cultura , Fungos/classificação , Fungos/isolamento & purificação , Humanos , Micoses/microbiologia , Manejo de Espécimes , Coloração e Rotulagem , Fatores de Tempo
7.
Medicine (Baltimore) ; 95(6): e2473, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26871771

RESUMO

One of the main concerns in psychiatric care is safety related to drug management. Pharmacogenetics provides an important tool to assess causes that may have contributed the adverse events during psychiatric therapy. This study illustrates the potential of pharmacogenetics to identify those patients for which pharmacogenetic-guided therapy could be appropriate. It aimed to investigate CYP2D6 genotype in our psychiatric population to assess the value of introducing pharmacogenetics as a primary improvement for predicting side effects.A broad series of 224 psychiatric patients comprising psychotic disorders, depressive disturbances, bipolar disorders, and anxiety disorders was included. The patients were genotyped with the AmpliChip CYP450 Test to analyzing 33 allelic variants of the CYP2D6 gene.All bipolar patients with poor metabolizer status showed maniac switching when CYP2D6 substrates such as selective serotonin reuptake inhibitors were prescribed. No specific patterns were identified for adverse events for other disorders.We propose to utilize pharmacogenetic testing as an intervention to aid in the identification of patients who are at risk of developing affective switching in bipolar disorder treated with selective serotonin reuptake inhibitors, CYP2D6 substrates, and inhibitors.


Assuntos
Antidepressivos de Segunda Geração/efeitos adversos , Transtorno Bipolar/tratamento farmacológico , Citocromo P-450 CYP2D6/genética , Genótipo , Inibidores Seletivos de Recaptação de Serotonina/efeitos adversos , Antidepressivos de Segunda Geração/uso terapêutico , Transtorno Bipolar/enzimologia , Transtorno Bipolar/genética , Citocromo P-450 CYP2D6/metabolismo , Marcadores Genéticos , Humanos , Segurança do Paciente , Melhoria de Qualidade , Inibidores Seletivos de Recaptação de Serotonina/uso terapêutico , Resultado do Tratamento
10.
Int J Syst Evol Microbiol ; 65(7): 2337-2340, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25888545

RESUMO

The species Seliberia stellata was described in 1963 and the name validly published in 1980. Its type strain, INMI N-9(T), was deposited in the VKM collection by one of the authors reporting its 5S rRNA gene sequence. Based on the analysis of this sequence, the currently distributed strains VKM B-1340 and CECT 7960 are not the original type strain of Seliberia stellata. A 16S rRNA gene sequence analysis of strain CECT 7960 had previously shown that this strain belongs to the species Bradyrhizobium betae, and this result was confirmed in the present paper by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) MS analysis for both CECT 7960 and VKM B-1340. Therefore, we propose that the Judicial Commission consider the following. (1) That the organism currently deposited as VKM B-1340 and CECT 7960 be recognized as a member of the species Bradyrhizobium betae. (2) That the organism deposited as VKM B-1340 and CECT 7960 does not represent the type strain of the species Seliberia stellata. (3) To place the species name Seliberia stellataAristovskaya and Parinkina 1963 (Approved Lists 1980) on the list of rejected names if a suitable replacement strain, or a neotype, cannot be found within two years of publication of this Request (Rule 18c). (4) To place the genus name SeliberiaAristovskaya and Parinkina 1963 (Approved Lists 1980) on the list of rejected names (Recommendation 20d) if a suitable replacement type strain or a neotype for the type species of the genus SeliberiaAristovskaya and Parinkina 1963 (Approved Lists 1980) is not identified as indicated in point (3).


Assuntos
Hyphomicrobiaceae/classificação , Filogenia , DNA Bacteriano/genética , RNA Ribossômico 16S/genética , RNA Ribossômico 5S/genética , Análise de Sequência de DNA , Terminologia como Assunto
11.
Toxicol Lett ; 234(2): 99-109, 2015 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-25677510

RESUMO

Nephrotoxicity limits the therapeutic efficacy of the antineoplastic drug cisplatin. Due to dosage adjustment and appropriate monitoring, most therapeutic courses with cisplatin produce no or minimal kidney damage. However, we studied whether even sub-nephrotoxic dosage of cisplatin poses a potential risk for the kidneys by predisposing to acute kidney injury (AKI), specifically by lowering the toxicity threshold for a second nephrotoxin. With this purpose rats were treated with a single sub-nephrotoxic dosage of cisplatin (3mg/kg, i.p.) and after two days, with a sub-nephrotoxic regime of gentamicin (50mg/kg/day, during 6 days, i.p.). Control groups received only one of the drugs or the vehicle. Renal function and renal histology were monitored throughout the experiment. Cisplatin treatment did not cause any relevant functional or histological alterations in the kidneys. Rats treated with cisplatin and gentamicin, but not those under single treatments, developed an overt renal failure characterized by both renal dysfunction and massive tubular necrosis. In addition, the urinary excretion of fumarylacetoacetase was increased in cisplatin-treated animals at subtoxic doses, which might be exploited as a cisplatin-induced predisposition marker. In fact, the urinary level of fumarylacetoacetase prior to the second nephrotoxin correlated with the level of AKI triggered by gentamicin in predisposed animals.


Assuntos
Injúria Renal Aguda/induzido quimicamente , Antineoplásicos/toxicidade , Cisplatino/toxicidade , Hidrolases/urina , Rim/efeitos dos fármacos , Injúria Renal Aguda/enzimologia , Injúria Renal Aguda/patologia , Injúria Renal Aguda/fisiopatologia , Injúria Renal Aguda/urina , Animais , Biomarcadores/urina , Modelos Animais de Doenças , Gentamicinas , Humanos , Rim/enzimologia , Rim/patologia , Rim/fisiopatologia , Necrose Tubular Aguda/induzido quimicamente , Necrose Tubular Aguda/enzimologia , Necrose Tubular Aguda/urina , Masculino , Ratos Wistar , Medição de Risco , Fatores de Risco , Fatores de Tempo , Regulação para Cima
12.
Pharmacogenomics ; 15(7): 997-1006, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24956253

RESUMO

AIM: Antiretroviral treatment implies a high cost to the healthcare system. The aim of this study was to evaluate the clinical and economic impact of efavirenz (EFV) dose adjustment by monitoring plasma concentrations and pharmacogenetic analysis of the 516G>T CYP2B6 polymorphism. MATERIALS & METHODS: One hundred and ninety HIV patients treated with EFV were studied. Plasma EFV concentrations were measured by HPLC with ultraviolet detection, and pharmacogenetic analysis was performed by Real Time (RT)-PCR. RESULTS: One hundred and ninety patients initially treated with a standard dose of EFV (600 mg/day) were studied. In 31 (16.3%) patients, EFV dose was reduced. A total of 87.1% of patients were heterozygous/homozygous carriers (GT/TT). CD4(+) count increased while the minimum steady-state plasma concentration and adverse effects decreased significantly after dose adjustment. Considering only the dose reduction, the adjustments accounted for a saving of 43,539 €/year. CONCLUSION: The individualization of EFV dosage guided by genotyping 516G>T CYP2B6 and therapeutic drug monitoring could increase the efficiency of EFV use in antiretroviral treatment.


Assuntos
Benzoxazinas/administração & dosagem , Citocromo P-450 CYP2B6/genética , Infecções por HIV/genética , Adulto , Alcinos , Fármacos Anti-HIV/administração & dosagem , Fármacos Anti-HIV/farmacocinética , Benzoxazinas/farmacocinética , Análise Custo-Benefício , Ciclopropanos , Monitoramento de Medicamentos , Feminino , Genótipo , HIV/efeitos dos fármacos , Infecções por HIV/tratamento farmacológico , Infecções por HIV/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Farmacogenética
13.
Rev Esp Quimioter ; 26(3): 193-7, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24080884

RESUMO

BACKGROUND: Recently, bacterial identification by MALDI-TOF MS has acquired a high relevance in terms of speed and reliability. Conventional mycological identification has some disadvantages: it is frequently slow, reliability is sometimes low, and an extensive experience is required. The risk population for fungal infections, and therefore their clinical significance has progressively increased in recent years. METHODS: 153 yeast and mould clinical isolates were analyzed by MALDI-TOF MS and conventional identification. When both methods were discrepant to the genus or species level, ITS-2 sequencing was performed. Results. The correlation in yeasts identification between conventional identification methods and MALDI-TOF MS was extremely high (99.2% to the species level and 100% to the genus level). The only discrepancy was checked by ITS-2 sequencing and confirmed the MALDI-TOF identification. The correlation in moulds identification was more heterogeneous. 68.7% of the isolates showed correlation at least to the genus level and 40.6% to the species level. Therefore, the correlation between conventional identification and MALDI-TOF MS in fungal identification was, in whole, 87% to the species level, and 93.5% to the genus level. CONCLUSIONS: Identification of fungi by MALDI-TOF MS is reliable and shows potential advantages over conventional identification methods.


Assuntos
Fungos/química , Micoses/microbiologia , Interpretação Estatística de Dados , Humanos , Técnicas Microbiológicas , Reprodutibilidade dos Testes , Especificidade da Espécie , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Leveduras/química
14.
Syst Appl Microbiol ; 36(8): 565-71, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24168963

RESUMO

Genus Bradyrhizobium includes slow growing bacteria able to nodulate different legumes as well as species isolated from plant tumours. The slow growth presented by the members of this genus and the phylogenetic closeness of most of its species difficults their identification. In the present work we applied for the first time Matrix-Assisted Laser Desorption Ionization-Time-of-Flight Mass Spectrometry (MALDI-TOF MS) to the analysis of Bradyrhizobium species after the extension of MALDI Biotyper 2.0 database with the currently valid species of this genus. With this methodology it was possible to identify strains belonging to phylogenetically closely related species of genus Bradyrhizobium allowing the discrimination among species with rrs gene identities higher than 99%. The application of MALDI-TOF MS to strains isolated from nodules of different Lupinus species in diverse geographical locations allowed their correct identification when comparing with the results of rrs gene and ITS analyses. The nodulation of Lupinus gredensis, an endemic species of the west of Spain, by B. canariense supports the European origin of this species.


Assuntos
Técnicas Bacteriológicas/métodos , Bradyrhizobium/química , Bradyrhizobium/classificação , Lupinus/microbiologia , Nódulos Radiculares de Plantas/microbiologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Bradyrhizobium/isolamento & purificação , Análise por Conglomerados , Filogenia , Análise de Sequência de DNA , Espanha
15.
Syst Appl Microbiol ; 36(8): 560-4, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24120265

RESUMO

Chicha is a drink prepared in several Andean countries from Inca's times by maize fermentation. Currently this fermentation is carried out in familiar artesanal "chicherías" that make one of the most known types of chicha, the "chicha de jora". In this study we isolate and identify the yeasts mainly responsible of the fermentation process in this type of chicha in 10 traditional "chicherías" in Cusco region in Peru. We applied by first time MALDI-TOF MS analysis for the identification of yeast of non-clinic origin and the results showed that all of yeast strains isolated belong to the species Saccharomyces cerevisiae. These results agree with those obtained after the analysis of the D1/D2 and 5.8S-ITS regions. However the chicha strains have a phenotypic profile that differed in more than 40% as compared to that of current S. cerevisiae strains. To the best of our knowledge this is the first report concerning the yeasts involved in chicha fermentation.


Assuntos
Bebidas Alcoólicas/microbiologia , Saccharomyces cerevisiae/classificação , Saccharomyces cerevisiae/isolamento & purificação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Fermentação , Técnicas Microbiológicas , Dados de Sequência Molecular , Técnicas de Tipagem Micológica , Peru , Filogenia , RNA Ribossômico 5,8S/genética , Saccharomyces cerevisiae/química , Saccharomyces cerevisiae/genética , Análise de Sequência de DNA , Zea mays/metabolismo , Zea mays/microbiologia
16.
Rev. esp. quimioter ; 26(3): 193-197, sept. 2013. tab
Artigo em Espanhol | IBECS | ID: ibc-115561

RESUMO

Introducción. En los últimos años, la identificación bacteriana mediante espectrometría de masas (EM) MALDI-TOF ha adquirido progresivamente importancia, dada su rapidez y fiabilidad. La identificación micológica convencional tiene algunos inconvenientes, como son su lentitud, su baja fiabilidad en algunos casos, y el hecho de que para algunos procedimientos es imprescindible la intervención de personas con amplia experiencia. Sin embargo, la población de riesgo para infecciones fúngicas, y por tanto su importancia clínica, han aumentado en los últimos años. Material y métodos. Se identificaron 153 aislamientos de hongos filamentosos y de levaduras mediante métodos convencionales y mediante EM MALDI-TOF. Cuando se dieron discrepancias entre ambos, éstas fueron dilucidadas mediante secuenciación del segmento ITS-2. Resultados. La correlación entre los métodos convencionales y la EM MALDI-TOF para la identificación de levaduras fue muy alta (99,2% a nivel de especie y 100% al de género). La única discrepancia que debió ser comprobada mediante secuenciación, corroboró el resultado obtenido con la EM. La correlación fue más heterogénea en el caso de los hongos filamentosos. 68,7% de los aislamientos mostraron correlación al menos al nivel de género, y 40,6% al nivel de especie. En conjunto, la correlación entre identificación convencional y MALDI-TOF para la identificación de hongos fue del 87% al nivel de especie, y del 93,5% a nivel de género. Conclusiones. La identificación de hongos mediante EM MALDI-TOF es fiable, y presenta ventajas potenciales con respecto a los métodos convencionales (AU)


Background. Recently, bacterial identification by MALDI-TOF MS has acquired a high relevance in terms of speed and reliability. Conventional mycological identification has some disadvantages: it is frequently slow, reliability is sometimes low, and an extensive experience is required. The risk population for fungal infections, and therefore their clinical significance has progressively increased in recent years. Methods. 153 yeast and mould clinical isolates were analyzed by MALDI-TOF MS and conventional identification. When both methods were discrepant to the genus or species level, ITS-2 sequencing was performed. Results. The correlation in yeasts identification between conventional identification methods and MALDI-TOF MS was extremely high (99.2% to the species level and 100% to the genus level). The only discrepancy was checked by ITS-2 sequencing and confirmed the MALDI-TOF identification. The correlation in moulds identification was more heterogeneous. 68.7% of the isolates showed correlation at least to the genus level and 40.6% to the species level. Therefore, the correlation between conventional identification and MALDI-TOF MS in fungal identification was, in whole, 87% to the species level, and 93.5% to the genus level. Conclusions. Identification of fungi by MALDI-TOF MS is reliable and shows potential advantages over conventional identification methods (AU)


Assuntos
Humanos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Fungos/isolamento & purificação , Micoses/diagnóstico , Leveduras/isolamento & purificação , Sensibilidade e Especificidade , Sequenciamento de Nucleotídeos em Larga Escala , Técnicas de Tipagem Micológica
17.
Electrophoresis ; 34(17): 2473-83, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23784626

RESUMO

Urine is a suitable biological fluid to look for markers of physiological and pathological processes, including renal and nonrenal diseases. In addition, it is an optimal body sample for diagnosis, because it is easily obtained without invasive procedures and can be sampled in large quantities at almost any time. Rats are frequently used as a model to study human diseases, and rat urine has been analyzed to search for disease biomarkers. The normal human urinary proteome has been studied extensively, but the normal rat urinary proteome has not been studied in such depth. In light of this, we were prompted to analyze the normal rat urinary proteome using three complementary proteomics platforms: SDS-PAGE separation, followed by LC-ESI-MS/MS; 2DE, followed by MALDI-TOF-TOF and 2D-liquid chromatography-chromatofocusing, followed by LC-ESI-Q-TOF. A total of 366 unique proteins were identified, of which only 5.2% of unique proteins were identified jointly by the three proteomics platforms used. This suggests that simultaneous proteomics techniques provide complementary and nonredundant information. Our analysis affords the most extensive rat urinary protein database currently available and this may be useful in the study of renal physiology and in the search for biomarkers related to renal and nonrenal diseases.


Assuntos
Biomarcadores/urina , Proteinúria/urina , Proteoma/análise , Proteômica/métodos , Animais , Biomarcadores/química , Cromatografia Líquida/métodos , Eletroforese em Gel Bidimensional/métodos , Feminino , Concentração de Íons de Hidrogênio , Espectrometria de Massas/métodos , Proteínas/química , Proteínas/classificação , Proteoma/química , Ratos , Ratos Wistar
18.
Toxicology ; 304: 83-91, 2013 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-23261757

RESUMO

Drug nephrotoxicity is a serious health and economic problem worldwide. Rats can be acutely sensitized to acute kidney injury (AKI) by subnephrotoxic treatments with potentially nephrotoxic drugs. Acquired sensitization to AKI poses a silent risk impossible to diagnose pre-emptively with the technology available at the clinical level. Herein, we hypothesized whether a chronic, subnephrotoxic insult to the kidneys might result in chronically acquired sensitization to AKI, and whether chronic sensitization might be detected through specific urinary markers. To this end, rats were treated with a subtoxic dosage of the experimental nephrotoxin uranyl nitrate (UN) in the drinking water for 21 weeks, or plain water (as control), and then with low-dose gentamicin for 7 days. Renal function and renal tissue damage were evaluated through the experiment. The mild renal damage caused by gentamicin was markedly magnified in rats having received UN chronically, which was evident both at the functional and histological level. Four proteins, namely albumin, hemopexin, transferrin and vitamin D binding protein were increased in the urine in temporal association with the appearance of chronic predisposition. Although further studies are necessary, our results suggest that these proteins might be potentially used as markers of hidden, chronic predisposition to gentamicin nephrotoxicity, in order to appropriately and pre-emptively stratify and handle individuals according to their specific risk in the long term, and to conveniently optimize their life conditions or additional clinical procedures or treatments that might trigger the disease. This might reduce AKI incidence and severity and the associated costs.


Assuntos
Injúria Renal Aguda/induzido quimicamente , Antibacterianos/toxicidade , Gentamicinas/toxicidade , Nitrato de Uranil/toxicidade , Injúria Renal Aguda/fisiopatologia , Albuminúria/induzido quimicamente , Animais , Antibacterianos/administração & dosagem , Biomarcadores/urina , Suscetibilidade a Doenças , Gentamicinas/administração & dosagem , Hemopexina/urina , Masculino , Ratos , Ratos Sprague-Dawley , Índice de Gravidade de Doença , Transferrina/urina , Nitrato de Uranil/administração & dosagem , Proteína de Ligação a Vitamina D/urina
19.
Enferm. infecc. microbiol. clín. (Ed. impr.) ; 30(10): 597-601, dic. 2012. tab
Artigo em Espanhol | IBECS | ID: ibc-106381

RESUMO

Objetivo. La espectrometría de masas (EM) MALDI-TOF se ha convertido en un recurso de referencia para la identificación de microorganismos en los servicios de microbiología clínica. No obstante, los datos relativos a algunos grupos de microorganismos son todavía controvertidos. En el presente estudio se ha determinado la fiabilidad de la EM MALDI-TOF para la identificación de aislamientos clínicos de bacterias anaerobias, en comparación con técnicas bioquímicas convencionales, y usando como referencia en caso de discrepancias las secuenciación de ARNr 16S.Material y métodos Se analizaron 126 aislamientos clínicos de bacterias anaerobias mediante el sistema API 20A (bioMérieux, Marcy l’Étoile, Francia) y mediante EM MALDI-TOF (Autoflex II, Bruker Daltonics, Alemania), utilizando la base de datos BioTyper 2.0 (Bruker Daltonics, Alemania). Cuando se produjeron discrepancias, o la EM MALDI-TOF no fue capaz de identificar microorganismo alguno, se usó como método de identificación de referencia la secuenciación del ARNr 16S.ResultadosEl método bioquímico y la EM MALDI-TOF coincidieron, a nivel de especie, en el 60,9% de los casos, y solo a nivel de género en el 20,3%. De las 48 identificaciones discrepantes, la secuenciación respaldó la identificación por EM MALDI-TOF a nivel de especie en 32 casos (66,7%), y a nivel de género en 8 (16,7%). Dicha secuenciación apoyó la identificación bioquímica a nivel de especie solamente en 2 casos (..) (AU)


Aim of the study: MALDI-TOF mass spectrometry (MS) is becoming a major resource in the Clinical Microbiology laboratory. Results on some groups of microorganisms are still controversial. We have studied the reliability of MALDI-TOF MS for the identification of anaerobic clinical isolates was studied compared to conventional biochemical methods, with rRNA 16S sequencing being used as a reference when discrepancies arose. Material and methods: A total of 126 anaerobic bacteria clinical isolates were studied by using API20Akits (bioMérieux, Marcy l’Étoile, France) and MALDI-TOF MS (Autoflex II, Bruker Daltonics, Germany), and using the data library BioTyper 2.0 (Bruker Daltonics, Germany). When discrepancies arose, or MALDI-TOFMS was not able to identify any microorganism, rRNA 16S sequencing was used as the reference standard. Results: The biochemical method and MALDI-TOF MS agreed in identifying 60.9% of isolates at species level, and 20.3% of isolates at genus level. Among the 48 discrepancies observed, rRNA 16S sequencing (..) (AU)


Assuntos
Humanos , Espectrometria de Massas , Bactérias Anaeróbias/isolamento & purificação , Infecções Bacterianas/microbiologia , Sensibilidade e Especificidade , Técnicas Bacteriológicas/métodos
20.
Artigo em Espanhol | IBECS | ID: ibc-104136

RESUMO

La espectrometría de masas (EM) matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) se ha impuesto rápidamente en numerosos Servicios de Microbiología Clínica como un nuevo recurso tecnológico. Su utilidad en la identificación bacteriana está contrastada, aunque existen todavía dudas respecto a la identificación de grupos bacterianos concretos y de otros microorganismos, como los hongos filamentosos. Existen además otras diversas aplicaciones potenciales de esta tecnología en Microbiología Clínica, que están empezando a desarrollarse. En esta revisión se resumen los datos existentes respecto a la identificación de diferentes grupos de microorganismos, incluyendo aquellas que han planteado mayores problemas, como micobacterias, anaerobios y hongos filamentosos. Se analizan también sus aplicaciones en diagnóstico directo sobre muestra, su repercusión en la consideración como patógenos de diferentes microorganismos, y sus potenciales aplicaciones epidemiológicas. Finalmente, se resumen también los estudios existentes sobre su uso potencial en la determinación de sensibilidad a antimicrobianos, y su potencial utilización usando como sustrato amplificados génicos en lugar de extractos proteicos de los microorganismos (AU)


Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) is rapidly becoming a new routine resource in Clinical Microbiology laboratories. Its usefulness for bacterial identification is now generally accepted, although there is still some reluctance as regards specific bacterial groups and some other microorganisms, such as moulds. There are other potential applications of this technology in Clinical Microbiology, which are beginning to be developed. A review is presented on the current data on the identification of microorganisms, including the most problematic groups, such as mycobacteria, anaerobic bacteria and moulds. We also analyse its applications for direct sample (..) (AU)


Assuntos
Humanos , Proteômica/métodos , /métodos , Técnicas Microbiológicas/métodos , Espectrometria de Massas/métodos , Testes de Sensibilidade Microbiana/métodos
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