Methicillin-resistant Staphylococcus aureus bacteremia:epidemiology, outcome, and laboratory characteristics in a tertiary referral center in the UK

Objective: To analyze the recent epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) bacteremia in a UK tertiary referral center. Methods: We collected epidemiological and laboratory data on all cases of MRSA bacteremia from September 1, 2005 to December 31, 2007. Results: There were 195 clinically significant episodes. Most were hospital-acquired. Only one episode occurred in patients without a history of hospital admission in the previous 12 months. An intravascular device was the most common focus of infection (37%), with no identifiable source found in 35% of episodes. Twenty-eight percent of patients died within 30 days of bacteremia. Mortality was significantly higher in the absence of an identifiable focus. Failure to include an antibiotic active against MRSA in the empirical treatment was only significantly associated with death in patients showing signs of hemodynamic instability (p < 0.001). No isolates had a minimum inhibitory concentration to vancomycin above 1.5. mg/l and no heteroresistance to glycopeptide antibiotics (heteroresistant vancomycin-intermediate Staphylococcus aureus; hVISA) was detected. All isolates were sensitive to daptomycin, tigecycline, and linezolid. Conclusions: Despite improvement in infection control measures, medical devices remain the most common source of infection. Inappropriate empirical antibiotic usage is associated with a poor outcome in patients with signs of severe sepsis. Susceptibility to glycopeptides and newer antibiotics remains good. © 2010 International Society for Infectious Diseases.

An ethnopharmacological approach to multidrug -resistant Staphylococcus aureus: Evaluation of Italian plants used in the traditional healing of skin disease

One-third of botanical remedies from southern Italy are used to treat skin and soft tissue infections (SST's). Methicillin-resistant Staphylococcus aureus (MRSA), a common cause of SSTIs, is responsible for increased morbidity and mortality from infections. Therapeutic options are limited by antibiotic resistance. Many plants possess potent antimicrobial compounds for these disorders. Validation of traditional medical practices is important for the people who rely on medicinal plants. Moreover, identification of novel antibiotics and anti-pathogenic agents for MRSA is important to global healthcare.^ I took an ethnopharmacological approach to understand how Italian medicinal plants used for the treatment of SSTIs affect MRSA growth and virulence. My hypothesis was that plants used in folk remedies for SSTI would exhibit lower cytotoxicity and greater inhibition of bacterial growth, biofilm formation and toxin production in MRSA than plants used for remedies unrelated to the skin or for plants with no ethnomedical application. The field portion of my research was conducted in the Vulture-Alto Bradano area of southern Italy. I collected 104 plant species and created 168 crude extracts. In the lab, I screened samples for activity against MRSA in a battery of bioassays. Growth inhibition was analyzed using broth microtiter assays for determination of the minimum inhibitory concentration. Interference with quorum-sensing (QS) processes, which mediate pathogenicity, was quantified through RP-HPLC of δ-toxin production. Interference with biofilm formation and adherence was assessed using staining methods. The mammalian cytotoxicity of natural products was analyzed using MTT cell proliferation assay techniques.^ Although bacteriostatic activity was limited, extracts from six plants used in Italian folk medicine (Arundo donax, Ballota nigra, Juglans regia, Leopoldia comosa, Marrubium vulgare, and Rubus ulmifolius ) significantly inhibited biofilm formation and adherence. Moreover, plants used to treat SSTI demonstrated significantly greater anti-biofilm activity when compared to plants with no ethnomedical application. QSI activity was evident in 90% of the extracts tested and extracts from four plants ( Ballota nigra, Castanea saliva, Rosmarinus officinalis, and Sambucus ebulus) exhibited a significant dose-dependent response. Some of the plant remedies for SSTI identified in this study can be validated due to anti-MRSA activity.^

Molecular analysis of evolutionary relationships of Phaseolus dumosus with the gene pool of common bean

Valuable genetic variation for bean breeding programs is held within the common bean secondary gene pool which consists of Phaseolus albescens, P. coccineus, P. costaricensis, and P. dumosus. However, the use of close relatives for bean improvement is limited due to the lack of knowledge about genetic variation and genetic plasticity of many of these species. Characterisation and analysis of the genetic diversity is necessary among beans' wild relatives; in addition, conflicting phylogenies and relationships need to be understood and a hypothesis of a hybrid origin of P. dumosus needs to be tested. This thesis research was orientated to generate information about the patterns of relationships among the common bean secondary gene pool, with particular focus on the species Phaseolus dumosus. This species displays a set of characteristics of agronomic interest, not only for the direct improvement of common bean but also as a source of valuable genes for adaptation to climate change. Here I undertake the first comprehensive study of the genetic diversity of P. dumosus as ascertained from both nuclear and chloroplast genome markers. A germplasm collection of the ancestral forms of P. dumosus together with wild, landrace and cultivar representatives of all other species of the common bean secondary gene pool, were used to analyse genetic diversity, phylogenetic relationships and structure of P. dumosus. Data on molecular variation was generated from sequences of cpDNA loci accD-psaI spacer, trnT-trnL spacer, trnL intron and rps14-psaB spacer and from the nrDNA the ITS region. A whole genome DArT array was developed and used for the genotyping of P. dumosus and its closes relatives. 4208 polymorphic markers were generated in the DArT array and from those, 742 markers presented a call rate >95% and zero discordance. DArT markers revealed a moderate genetic polymorphism among P. dumosus samples (13% of polymorphic loci), while P. coccineus presented the highest level of polymorphism (88% of polymorphic loci). At the cpDNA one ancestral haplotype was detected among all samples of all species in the secondary genepool. The ITS region of P. dumosus revealed high homogeneity and polymorphism bias to P. coccineus genome. Phylogenetic reconstructions made with Maximum likelihood and Bayesian methods confirmed previously reported discrepancies among the nuclear and chloroplast genomes of P. dumosus. The outline of relationships by hybridization networks displayed a considerable number of interactions within and between species. This research provides compelling evidence that P. dumosus arose from hybridisation between P. vulgaris and P. coccineus and confirms that P. costaricensis has likely been involved in the genesis or backcrossing events (or both) in the history of P. dumosus. The classification of the specie P. persistentus was analysed based on cpDNA and ITS sequences, the results found this species to be highly related to P. vulgaris but not too similar to P. leptostachyus as previously proposed. This research demonstrates that wild types of the secondary genepool carry a significant genetic variation which makes this a valuable genetic resource for common bean improvement. The DArT array generated in this research is a valuable resource for breeding programs since it has the potential to be used in several approaches including genotyping, discovery of novel traits, mapping and marker-trait associations. Efforts should be made to search for potential populations of P. persistentus and to increase the collection of new populations of P. dumosus, P. albescens and P. costaricensis that may provide valuable traits for introgression into common bean and other Phaseolus crops.

Concentración de cationes (Ca2+, Mg2+, Fe2+3+, K+) en semillas de frijol negro (Phaseolus vulgaris L.) usando espectroscopia de absorción atómica

Se investiga la concentración de cationes (Ca,Fe,K,Mg),en el frijol negro,(Phaseolus vulgaris L. var. Tineco), usando un analizador de absorción atómica. Para tal efecto se procede a secar la muestra y luego se digiere en un medio acido. Se preparan las soluciones standard correspondiente a los cationes que se quiere investigar y luego se comparan estas concentraciones con las concentraciones de los iones en la muestra. El cálculo de la concentración (desconocida), de los cationes en la muestra, se hace por medio de simple aritmética. En este trabajo se reporta la cantidad en p. p. m. de los elementos arriba mencionados, presente en un gramo de la muestra original. Los resultados obtenidos fueron: Fe = 337.5 p. p. m. ; Ca = 150 p. p. m. ; Mg = 150 p. p. m. y K = 4X104p. p. m.

Antibacterial activity of the marine sponge Ircinia Campana collected at Punta Uva Limón against Staphylococcus Aureus

The sponges are simple multicellularorganisms; they inhabit in marine environments from the polar seas to the tropical waterswhere they are more abundant. These species are exposed to large populations of microbes, reason that explains their complex morphological and cellular defense mechanism, which are used by these organisms to fight against pathogens. The purpose of this study was to evaluate the antibacterial activity of the marine sponge Ircinia campana, whichinhabits in the south of the Caribbean coast of Costa Rica against  Sthapylococcus aureus gram-positive bacteria. Sampleswere collected in Punta Uva in Limónduring July of 2007. The active compounds were obtainedby extraction with acetone (crude extract); and subsequently, chromatographic extracts were obtained using fractions 1:4 hexane: ethyl acetate. The antibacterial activities of the different fractions, including the  crude extract were tested.Our results suggest a zone of inhibition of 14.60 ±0.25 mm for the crude extract and18.70±0.25mm for the most active fraction separated by chromatography. The metabolite responsible for the antibacterial activity was isolated by High Performance Liquid Chromatography (HPLC)and preliminarily characterized through ultraviolet (UV) and infrared (IR) spectroscopy.

Caracterización, evaluación preliminar y adaptabilidad de cuatro poblaciones de frijol común (Phaseolus vulgaris L.), evaluadas en cinco localidades de Ciudad Darío, Matagalpa; Postrera, 2013

En el estudio realizado en el municipio de Ciudad Dar í o, Matagalpa, é poca de postrera 2013, se sometieron cuatro variedades de frijol com ú n ( Phaseolus vulgaris L.), a los diferentes ambientes de las localidades El Moj ó n, El Rinc ó n, El Pital, Dulce Nombre de Jes ú s y la Picota donde se evaluaron dos variedades locales (Guanise ñ o, Rojo Maravilla), una aclimatada a la zona (Rojo seda), y una variedad mejorada (INTA sequia). Para el ensayo se establecieron cuatro parcelas de 10 m de largo por 6 m de ancho, en donde la parcela ú til tuvo una dimensi ó n 6 m de largo y 3 m de ancho, cabe mencionar que cada parcela estaba constituida de 5 surcos, de las que se evaluaron 5 plantas por surco para un total de 25 plantas por cada parcela, adem á s cada finca se consider ó una r é plica. Los objetivos planteados fueron 1) Caracterizar y evaluar preliminarmente mediante variables cuantitativas y cualitativas cuatro cultivares de frijol (dos criollas, una acriollada y una mejorada), en cinco localidades de Ciudad Dar í o, Matagalpa, en la é poca de postrera 2013 y 2) Determinar la adaptabilidad de cuatro variedades de frijol com ú n (dos criollas, una acriollada y una mejorada), en base a los rendimientos obtenidos por cada variedad en estudio. En el siguiente estudio se realiz ó un an á lisis de adaptabilidad el cual se llev ó a cabo por medio del an á lisis de regresi ó n lineal, adem á s se evalu ó el comportamiento de los caracteres cualitativos y cuantitativos para determinar la variabilidad gen é tica existentes en las diferentes variedades. En donde se demuestra que en los caracteres cualitativos no existi ó variabilidad dado que estos caracteres son gobernados por uno o pocos genes, en cambio se pudo observar que en los caracteres cuantitativos existe alta heterogeneidad entre los mismos dado que estos caracteres son influenciados por el ambiente, adem á s se conoci ó cu á l de las cuatro variedades en estudio present ó mejor adaptabilidad en las diferentes localidades, tambi é n se informa sobre la variedad por la que el productor debe optar para la siembra del cultivo antes mencionado

Diagnóstico de suelos en cultivos de café (Coffea arabica L.), maíz (Zea mays L.) y frijol (Phaseolus vulgaris L.), comunidad de Santa Julia, El Crucero, Managua

Uno de los parámetros de mayor importancia en la agricultura es el estado nutricional del suelo, así como el manejo de parte de los agricultores. En 2014, a través del programa para el Desarrollo Participativo Integral Rural (DEPARTIR)se efectuó un muestreo de suelos a profundidad de 25 cm en 34 lotes productivos de 17 unidades de producción, dedicadas a la producción de café (Coffea arabica), maíz ( Zea mays L.) y frijol (Phaseolusvulgaris L.) en la comunidad Santa Julia, El Crucero, Managua. Se analizaron los factores que condicionan el uso del suelo, a través de análisis de correlación, ANDEVA y análisis de correspondencia en N, P,K, Mg, Ca, MO, CO y pH. El N total,y altitudfueron correlacionadas. El 95% de las áreas presentaron 10.5 hectáreas, con predominancia decultivo de café. Lafertilización químicaes unapráctica común en las unidades productivasy cultivos estudiados. Los muestreos efectuados en las parcelas de maíz fueron homogéneos a lo interno y externo de las unidades productivas; no así en el cultivo de café. Las áreas de siembra, destino de la producción y la aplicación de fertilizantes químicos, presentaron una alta relación.

Adaptabilidad y estabilidad de cuatro variedades de frijol (Phaseolus vulgaris L.) en seis localidades de San Dionisio, Matagalpa, en dos ciclos agrícolas, postrera 2013 y primera 2014

El cultivo del Frijol ( Phaseolus vulgaris L.) es uno de los principales rubro s que producen los agricultores del Municipio de San Dionisio , Matagalpa, haciendo uso de diferentes variedades criollas y acriolladas persistentes en la zona. El objetivo de ésta investigación fue estudiar la variabilidad fenotípica y la adaptabilidad de t res variedades criollas ( Guaniseño, Rojo seda y Rack sequía) y una variedad acriollada (INTA Sequía) durante d os ciclos agrícolas ( Postrera 2013 y Primera 2014 ) . Para esto , se establecieron las parcelas experimentales en una finca por localidad (6), l as dimensiones de las parcelas fueron 5 m de ancho por 10 m de largo y la parcela útil consistió de 3 m de ancho y 6 m de longitud. Para la variabilidad fenotípica se analizaron variables cuantitativas de crecimiento y desarrollo de las plantas a través de medidas de tendencias estadísticas y para el análisis de adaptabilidad se analizó la estabilidad del rendimiento de las variedades en las dife rentes localidades por medio de Análisis de Varianza y de Regresión Lineal . Entre los resul tados obtenidos se enc ontró que todas las variedades criollas y acriolladas evaluadas presentaron mayor variabilidad fenotípica en el ciclo agrícola postrera 2013. Las variables emergencia, vaina s por planta, plantas cosechadas y rendimiento por parcela mostraron los coeficientes de variación más altos con valores que oscilan entre 17.96 y 56.01 , principalmente la variable rendimiento p or parcela que fue la que mostró los valores más altos. En cuan to a la adaptabilidad de las variedades , el modelo de regresión lineal no resultó apropiado para analizar los datos obtenidos en ambos ciclos , sin embargo , se pudo tener una idea de la estabilidad de las variedades a través de la Regresión de la Desviación en donde se encontró que la varied ad Rack Sequia fue la que mostró menor coeficiente de Desviación (8.22) lo que demuestra que sus rendimientos fueron los más estables a nivel de fincas y ciclos agrícolas por lo tanto, fue la mejor adaptada a los diferentes ambientes.

Inoculaci?n y establecimiento endof?tico de cepas de los hongos entomopat?genos beauveria bassiana, Metarhizium anisopliae y lecanicillium lecanii en plantas de fr?jol (Phaseolus vulgaris) [recurso electr?nico]

Los hongos end?fitos han recibido una creciente atenci?n en los a?os recientes, ya que han demostrado tener un potencial econ?mico en la agronom?a, debido a su posible uso como agentes de control biol?gico. El establecimiento endof?tico de hongos entomopat?genos en cultivos de inter?s, ofrece un campo amplio y prometedor para comprender las relaciones entre plantas y hongos y los mecanismos de defensa que estos ?ltimos pueden inducir en las plantas. El presente estudio busc? inocular y evaluar la colonizaci?n endof?tica de tres especies de hongos entomopat?genos en plantas de fr?jol (Phaseolus vulgaris). Para ello, se emplearon dos cepas de cada especie, una comercial y la otra de la colecci?n de entomopat?genos del Centro Internacional de Agricultura Tropical; cada cepa constituy? un tratamiento individual. A partir de la siembra y desarrollo de las cepas, se generaron in?culos de 108 conidios/mL que fueron inoculados en las plantas de fr?jol Diacol, variedad Calima. Se encontr? que Beauveria bassiana, Metarhizium anisopliae y Lecanicillium lecanii colonizaron individualmente las hojas de fr?jol. Los porcentajes de colonizaci?n promedio para las cepas de la colecci?n del CIAT fueron de 16% (?= 18,84) para B. bassiana, 13% (?= 4,96) en M. anisopliae y 4% (?= 4,64) para L. lecanii. Con respecto a las cepas comerciales, estos valores fueron 4% (?= 4,64) para B. bassiana; 26% (?= 32) para M. anisopliae y 44% (?= 13,02) para L. lecanii. Todas las cepas de hongos inoculadas pudieron ser recuperadas de tejido vegetal, lo que sugiere su posible uso como agentes de control biol?gico al menos para el cultivo del fr?jol.

Caracterização fisio-molecular de cultivares de feijão (Phaseolus vulgaris L.) em resposta à deficiência hídrica

The identification of genotypes for drought tolerance has a great importance in breeding programs. The aim of this study was to characterize genotypes of beans in response to drought tolerance in different reproductive stages through physiologic, agronomic and molecular analysis. The experiment was conducted in greenhouse, using a randomized block design with four replicates; 10 cultivars: ANFC 9, ANFP 110, BRS Esplendor, BRSMG Realce, IPR Siriri, IPR Tangará, IPR Tuiuiu, IPR Uirapuru, IAC Imperador and IAC Milênio under two conditions of irrigation: plants irrigated during their entire life cycle, and plants under irrigation suppression in the reproductive stage (R7) until 16% of field capacity, when the irrigation was restored. In the last four days of stress, the gas exchanges were analyzed, and in the last day of stress was analyzed the percentage of closed stomata in the abaxial surface of the leaves, collected in different times of the day (9h, 12h, 15h and 18h). Additionally, plant samples were collected for the following analysis: fresh and dry mass of leaves, stems and legumes, and proline content in leaves and roots. The plants were harvested at the physiological maturity and the yield components and grain yield were determined. In addition, in order to identify polymorphisms in the sequences of promoters and genes related to drought, seven pairs of primers were tested on the group of genotypes. The drought susceptibility indexes (ISS) ranged from 0.65 to 1.10 in the group of genotypes, which the lowest values observed were for IAC Imperador (0.65) and BRS Esplendor (0.87), indicating the ability of these two genotypes to maintain grain yield under water stress condition. All genotypes showed reduction in yield components under water stress. IAC Imperador (43.4%) and BRS Esplendor (60.6%) had the lowest reductions in productivity and kept about 50% of the stomata closed during all the different times evaluated at last day of irrigation suppression. IAC Imperador showed greater water use efficiency and CO2 assimilation rate under drought stress. IPR Tuiuiú, IPR Tangará and IAC Imperador had the highest proline concentrations in the roots. Under water stress condition, there was a strong positive correlation (0.696) between the percentage of stomata closed with the number of grains per plant (0.696) and the fresh mass of leaves (0.731), the maximum percentage of stomata closed 73.71% in water stress. The accumulation of proline in the root was the character that most contributed to the divergence between the genotypes under water deficit, but not always the genotypes that have accumulated more proline were the most tolerant. The polymorphisms in DNA of coding and promoting sequences of transcription factors studied in this experiment did not discriminate tolerant genotypes from the sensitive ones to water stress.

Proteomic and phosphoproteomic analysis of polyethylene glycol-induced osmotic stress in root tips of common bean (Phaseolus vulgaris L.)

Previous studies have shown that polyethylene glycol (PEG)-induced osmotic stress (OS) reduces cell-wall (CW) porosity and limits aluminium (Al) uptake by root tips of common bean (Phaseolus vulgaris L.). A subsequent transcriptomic study suggested that genes related to CW processes are involved in adjustment to OS. In this study, a proteomic and phosphoproteomic approach was applied to identify OS-induced protein regulation to further improve our understanding of how OS affects Al accumulation. Analysis of total soluble proteins in root tips indicated that, in total, 22 proteins were differentially regulated by OS; these proteins were functionally categorized. Seventy-seven per- cent of the total expressed proteins were involved in metabolic pathways, particularly of carbohydrate and amino acid metabolism. An analysis of the apoplastic proteome revealed that OS reduced the level of five proteins and increased that of seven proteins. Investigation of the total soluble phosphoproteome suggested that dehydrin responded to OS with an enhanced phosphorylation state without a change in abundance. A cellular immunolocalization analysis indicated that dehydrin was localized mainly in the CW. This suggests that dehydrin may play a major protective role in the OS-induced physical breakdown of the CW structure and thus maintenance of the reversibility of CW extensibility during recovery from OS. The proteomic and phosphoproteomic analyses provided novel insights into the complex mechanisms of OS-induced reduction of Al accumulation in the root tips of common bean and highlight a key role for modification of CW structure.

Physiological and molecular analysis of the interaction between aluminium toxicity and drought stress in common bean (Phaseolus vulgaris)

Aluminium (Al) toxicity and drought are two major factors limiting common bean (Phaseolus vulgaris) production in the tropics. Short-term effects of Al toxicity and drought stress on root growth in acid, Al-toxic soil were studied, with special emphasis on Al-drought interaction in the root apex. Root elongation was inhibited by both Al and drought. Combined stresses resulted in a more severe inhibition of root elongation than either stress alone. This result was different from the alleviation of Al toxicity by osmotic stress (-0.60 MPa polyethylene glycol) in hydroponics. However, drought reduced the impact of Al on the root tip, as indicated by the reduction of Al-induced callose formation and MATE expression. Combined Al and drought stress enhanced up-regulation of ACCO expression and synthesis of zeatin riboside, reduced drought-enhanced abscisic acid (ABA) concentration, and expression of NCED involved in ABA biosynthesis and the transcription factors bZIP and MYB, thus affecting the regulation of ABA-dependent genes (SUS, PvLEA18, KS-DHN, and LTP) in root tips. The results provide circumstantial evidence that in soil, drought alleviates Al injury, but Al renders the root apex more drought-sensitive, particularly by impacting the gene regulatory network involved in ABA signal transduction and cross-talk with other phytohormones necessary for maintaining root growth under drought.

Alteration of cell-wall porosity is involved in osmotic stress-induced enhancement of aluminium resistance in common bean (Phaseolus vulgaris L.)

Aluminium (Al) toxicity and drought are the two major abiotic stress factors limiting common bean production in the tropics. Using hydroponics, the short-term effects of combined Al toxicity and drought stress on root growth and Al uptake into the root apex were investigated. In the presence of Al stress, PEG 6000 (polyethylene glycol)-induced osmotic (drought) stress led to the amelioration of Al-induced inhibition of root elongation in the Al-sensitive genotype VAX 1. PEG 6000 (>> PEG 1000) treatment greatly decreased Al accumulation in the 1 cm root apices even when the roots were physically separated from the PEG solution using dialysis membrane tubes. Upon removal of PEG from the treatment solution, the root tips recovered from osmotic stress and the Al accumulation capacity was quickly restored. The PEG-induced reduction of Al accumulation was not due to a lower phytotoxic Al concentration in the treatment solution, reduced negativity of the root apoplast, or to enhanced citrate exudation. Also cell-wall (CW) material isolated from PEG-treated roots showed a low Al-binding capacity which, however, was restored after destroying the physical structure of the CW. The comparison of the Al(3+), La(3+), Sr(2+), and Rb(+) binding capacity of the intact root tips and the isolated CW revealed the specificity of the PEG 6000 effect for Al. This could be due to the higher hydrated ionic radius of Al(3+) compared with other cations (Al(3+) >> La(3+) > Sr(2+) > Rb(+)). In conclusion, the results provide circumstantial evidence that the osmotic stress-inhibited Al accumulation in root apices and thus reduced Al-induced inhibition of root elongation in the Al-sensitive genotype VAX 1 is related to the alteration of CW porosity resulting from PEG 6000-induced dehydration of the root apoplast.