Crystallization and preliminary X-ray diffraction studies of Staphylococcus aureus homoserine dehydrogenase

Staphylococcus aureus is a Gram-positive nosocomial pathogen. The prevalence of multidrug-resistant S. aureus strains in both hospital and community settings makes it imperative to characterize new drug targets to combat S. aureus infections. In this context, enzymes involved in cell-wall maintenance and essential amino-acid biosynthesis are significant drug targets. Homoserine dehydrogenase (HSD) is an oxidoreductase that is involved in the reversible conversion of l-aspartate semialdehyde to l-homoserine in a dinucleotide cofactor-dependent reduction reaction. HSD is thus a crucial intermediate enzyme linked to the biosynthesis of several essential amino acids such as lysine, methionine, isoleucine and threonine.

Lipid coated mesoporous silica nanoparticles as an oral delivery system for targeting and treatment of intravacuolar Salmonella infections

Lipid coated mesoporous silica nanoparticle (L-MSN) were synthesized for oral delivery of ciprofloxacin for intracellular elimination of Salmonella pathogen. The particle size was found to be between 50-100 nm with a lipid coat of approximately 5 nm thickness. The lipid coating was achieved by sonication of liposomes with the MSN particles and evaluated by CLSMand FTIR studies. The L-MSN particles exhibited lower cytotoxicity compared to bare MSN particles. Ciprofloxacin, a fluoroquinolone antibiotic, loaded into the L-MSN particles showed enhanced antibacterial activity against free drug in in vitro assays. The lipid coat was found to aid in intravacuolar targeting of the drug cargo as observed by confocal microscopy studies. We also observed that a lower dose of antibiotic was sufficient to clear the pathogen from mice and increase their survivability using the L-MSN oral delivery system.

Successful treatment of biofilm infections using shock waves combined with antibiotic therapy

Many bacteria secrete a highly hydrated framework of extracellular polymer matrix on suitable substrates and embed within the matrix to form a biofilm. Bacterial biofilms are observed on many medical devices, endocarditis, periodontitis and lung infections in cystic fibrosis patients. Bacteria in biofilm are protected from antibiotics and >1,000 times of the minimum inhibitory concentration may be required to treat biofilm infections. Here, we demonstrated that shock waves could be used to remove Salmonella, Pseudomonas and Staphylococcus biofilms in urinary catheters. The studies were extended to a Pseudomonas chronic pneumonia lung infection and Staphylococcus skin suture infection model in mice. The biofilm infections in mice, treated with shock waves became susceptible to antibiotics, unlike untreated biofilms. Mice exposed to shock waves responded to ciprofloxacin treatment, while ciprofloxacin alone was ineffective in treating the infection. These results demonstrate for the first time that, shock waves, combined with antibiotic treatment can be used to treat biofilm infection on medical devices as well as in situ infections.

Asparagine requirement in Plasmodium berghei as a target to prevent malaria transmission and liver infections

The proteins of Plasmodium, the malaria parasite, are strikingly rich in asparagine. Plasmodium depends primarily on host haemoglobin degradation for amino acids and has a rudimentary pathway for amino acid biosynthesis, but retains a gene encoding asparagine synthetase (AS). Here we show that deletion of AS in Plasmodium berghei (Pb) delays the asexual-and liver-stage development with substantial reduction in the formation of ookinetes, oocysts and sporozoites in mosquitoes. In the absence of asparagine synthesis, extracellular asparagine supports suboptimal survival of PbAS knockout (KO) parasites. Depletion of blood asparagine levels by treating PbASKO-infected mice with asparaginase completely prevents the development of liver stages, exflagellation of male gametocytes and the subsequent formation of sexual stages. In vivo supplementation of asparagine in mice restores the exflagellation of PbASKO parasites. Thus, the parasite life cycle has an absolute requirement for asparagine, which we propose could be targeted to prevent malaria transmission and liver infections.

Intracellular demography and the dynamics of Salmonella enterica infections.

An understanding of within-host dynamics of pathogen interactions with eukaryotic cells can shape the development of effective preventive measures and drug regimes. Such investigations have been hampered by the difficulty of identifying and observing directly, within live tissues, the multiple key variables that underlay infection processes. Fluorescence microscopy data on intracellular distributions of Salmonella enterica serovar Typhimurium (S. Typhimurium) show that, while the number of infected cells increases with time, the distribution of bacteria between cells is stationary (though highly skewed). Here, we report a simple model framework for the intensity of intracellular infection that links the quasi-stationary distribution of bacteria to bacterial and cellular demography. This enables us to reject the hypothesis that the skewed distribution is generated by intrinsic cellular heterogeneities, and to derive specific predictions on the within-cell dynamics of Salmonella division and host-cell lysis. For within-cell pathogens in general, we show that within-cell dynamics have implications across pathogen dynamics, evolution, and control, and we develop novel generic guidelines for the design of antibacterial combination therapies and the management of antibiotic resistance.

Virulent Salmonella enterica infections can be exacerbated by concomitant infection of the host with a live attenuated S. enterica vaccine via Toll-like receptor 4-dependent interleukin-10 production with the involvement of both TRIF and MyD88.

During systemic disease in mice, Salmonella enterica grows intracellularly within discrete foci of infection in the spleen and liver. In concomitant infections, foci containing different S. enterica strains are spatially separated. We have investigated whether functional interactions between bacterial populations within the same host can occur despite the known spatial separation of the foci and independence of growth of salmonellae residing in different foci. In this study we have demonstrated that bacterial numbers of virulent S. enterica serovar Typhimurium C5 strain in mouse tissues can be increased by the presence of the attenuated aroA S. Typhimurium SL3261 vaccine strain in the same tissue. Disease exacerbation does not require simultaneous coinjection of the attenuated bacteria. SL3261 can be administered up to 48 hr after or 24 hr before the administration of C5 and still determine higher tissue numbers of the virulent bacteria. This indicates that intravenous administration of a S. enterica vaccine strain could potentially exacerbate an established infection with wild-type bacteria. These data also suggest that the severity of an infection with a virulent S. enterica strain can be increased by the prior administration of a live attenuated vaccine strain if infection occurs within 48 hr of vaccination. Exacerbation of the growth of C5 requires Toll-like receptor 4-dependent interleukin-10 production with the involvement of both Toll/interleukin-1 receptor-domain-containing adaptor inducing interferon-beta and myeloid differentiation factor 88.

Bacterial growth rate and host factors as determinants of intracellular bacterial distributions in systemic Salmonella enterica infections.

Bacteria of the species Salmonella enterica cause a range of life-threatening diseases in humans and animals worldwide. The within-host quantitative, spatial, and temporal dynamics of S. enterica interactions are key to understanding how immunity acts on these infections and how bacteria evade immune surveillance. In this study, we test hypotheses generated from mathematical models of in vivo dynamics of Salmonella infections with experimental observation of bacteria at the single-cell level in infected mouse organs to improve our understanding of the dynamic interactions between host and bacterial mechanisms that determine net growth rates of S. enterica within the host. We show that both bacterial and host factors determine the numerical distributions of bacteria within host cells and thus the level of dispersiveness of the infection.

Determinacion de frecuencia de politraumatismo oseo (ptos) en mascotas caninas en el hospital animal El Dorado

Ante la falta de información acerca de la frecuencia, diagnóstico, tratamiento y primeros auxilios de la mascota canina politraumatizada en nuestro país, y la falta de medios para la praxis ortopédica, se propuso este estudio de tesis; cuyo titulo es: Determinación de frecuencia de politraumatismo óseo con diagnostico radiológico en mascotas caninas en el Hospital Animal El Dorado. Para lo cual se procedió a establecer los siguientes objetivos: la evaluación de la frecuencia de politraumatismos óseos (PTO) atendidos en la clínica veterinaria El Dorado con diagnostico radiológico, para esto se realizó dicho estudio en un periodo de 3 meses, en el cual se estudiaron expedientes y recopilación de datos, con este estudio de tesis se logró comprobar que la mayor afectación por fracturas la sufren los canes de raza criolla, seguidos por los Pastores Alemanes, así mismo se determinó que el manejo de las mascotas es determinante y predisponente a que una mascota sufra una fractura. Se identificó que los animales menores de 2 años sufren mayormente fracturas. Se concluyó que debido a los altos costos de la praxis, muchos veterinarios no atienden a sus pacientes y también debido a la misma razón muchos dueños de mascotas optan por la eutanasia o simplemente dejan que el tiempo repare las fracturas. Este estudio pretende ser una herramienta que permita al estudiante de Medicina Veterinaria tener un mayor entendimiento acerca de los politraumatismos. Para el profesional una referencia. Se concluyó que la raza y edad de las mascotas, así mismo, como la época del año (invierno) son factores determinantes en el surgimiento de Politraumatismo óseo, los cuales se localizan con mayor frecuencia en mascotas a nivel cubito/radio, fémur, tibia y peroné, la terapia mayormente utilizada fue la conservadora mediante el uso de férulas seguida de la quirúrgica con el uso de cerclaje.

Intracellular demography and the dynamics of Salmonella enterica infections.

An understanding of within-host dynamics of pathogen interactions with eukaryotic cells can shape the development of effective preventive measures and drug regimes. Such investigations have been hampered by the difficulty of identifying and observing directly, within live tissues, the multiple key variables that underlay infection processes. Fluorescence microscopy data on intracellular distributions of Salmonella enterica serovar Typhimurium (S. Typhimurium) show that, while the number of infected cells increases with time, the distribution of bacteria between cells is stationary (though highly skewed). Here, we report a simple model framework for the intensity of intracellular infection that links the quasi-stationary distribution of bacteria to bacterial and cellular demography. This enables us to reject the hypothesis that the skewed distribution is generated by intrinsic cellular heterogeneities, and to derive specific predictions on the within-cell dynamics of Salmonella division and host-cell lysis. For within-cell pathogens in general, we show that within-cell dynamics have implications across pathogen dynamics, evolution, and control, and we develop novel generic guidelines for the design of antibacterial combination therapies and the management of antibiotic resistance.

La Facultad de Teología en una Iglesia llamada a ser “hospital de campaña”

En el mes de octubre del año pasado se realizó en la Facultad una Reunión de Claustro en la que participamos unos cuarenta profesores, reunidos para conversar e intercambiar ideas acerca de la entrevista que el P. Antonio Spadaro S.J. le hiciera al papa Francisco. Como los temas abordados allí fueron muchos, nos pareció oportuno señalar los que podían interesar de manera más directa a nuestra Facultad. En el diálogo tuvimos presente esa perspectiva, la de la posible recepción o incidencia entre nosotros de algunas de las afirmaciones hechas en esa entrevista. La conversación giró en torno a cuatro grandes campos temáticos: Eclesiológico; Pastoral-Moral; Espiritualidad; Diálogo fe-cultura. Haciéndome eco de esa rica experiencia, que seguramente se prolongará en el presente año académico, quisiera retener un párrafo de dicha entrevista, a partir del cual intentaré reflexionar manteniendo la misma perspectiva, a saber, la de su posible incidencia en la vida de nuestra Facultad...

Un modelo de programación por metas para la elaboración del contrato-programa de un hospital público

[ES] Proponemos un modelo de programación por metas para la estimación del plan de producción (case-mix) que debe reflejarse en el Contrato–Programa que suscriben anualmente los Hospitales Públicos y la Administración. Las variables de decisión son los volúmenes de actividad de cada servicio médico del hospital y los atributos son los indicadores básicos que se manejan al elaborar el Contrato-Programa: fi nanciación, número de altas, estancia media y peso de complejidad. Para resolver nuestro modelo empleamos la herramienta SOLVER de la hoja de cálculo EXCEL. La utilización de esta herramienta permite simular varios escenarios de una manera ágil, lo que es de gran ayuda para el estudio y discusión de las cantidades a contratar entre el Hospital y la Administración. El artículo finaliza con una breve presentación de los resultados obtenidos al aplicar nuestro modelo a un hospital de tamaño medio (118 camas) del Servicio Vasco de Salud.