Draft genome sequence and gene annotation of the uropathogenic bacterium Proteus mirabilis Pr2921

Here, we report the genome sequence of Proteus mirabilis Pr2921, a uropathogenic bacterium that can cause severe complicated urinary tract infections. After gene annotation, we identified two additional copies of ucaA, one of the most studied fimbrial protein genes, and other fimbriae related-proteins that are not present in P. mirabilis HI4320

Renin-Angiotensin-Aldosterone System Inhibitors, Statins, and Beta-Blockers in Diabetic Patients With Critical Limb Ischemia and Foot Lesions

Medical therapy for secondary prevention is known to be under-used in patients with peripheral artery disease (PAD). Few data are available on the subgroup with critical limb ischemia (CLI). Prescription of cardiovascular preventive therapies was recorded at discharge in a large, prospective cohort of patients admitted for treatment of CLI and foot lesions, stratified for coronary artery disease (CAD) diagnosis. All patients were followed up for at least 1 year. The primary endpoint was major adverse cardiovascular events (MACE). 618 patients were observed for a median follow-up of 981 days. Renin-angiotensin-aldosterone system (RAAS) inhibitors, statins, beta-blockers, and antithrombotic drugs were prescribed in 52%, 80%, 51%, and 99% of patients, respectively. However, only 43% of patients received optimal medical therapy (OMT), defined as the combination of RAAS inhibitor plus statin plus at least one antithrombotic drug. It was observed that the prescription of OMT was not affected by the presence of a CAD diagnosis. On the other hand, it was noticed that the renal function affected the prescription of OMT. OMT was independently associated with MACE (HR 0.688, 95%CI 0.475-0.995, P = .047) and, after propensity matching, also with all-cause mortality (HR 0.626, 95%CI 0.409-0.958, P = .031). Beta-blockers prescription was not associated with any outcome. In conclusion, patients with critical limb ischemia are under-treated with cardiovascular preventive therapies, irrespective of a CAD diagnosis. This has consequences on their prognosis

Fluoxetine and thioridazine inhibitefflux and attenuate crystalline biofilm formation by Proteusmirabilis

Proteus mirabilis forms extensive crystalline biofilms on indwelling urethral catheters that block urineflow and lead to serious clinical complications. The Bcr/CflA efflux system has previously been identified as important for development of P. mirabilis crystalline biofilms, highlighting the potential for efflux pump inhibitors (EPIs) to control catheter blockage. Here we evaluate the potential for drugs already used in human medicine (fluoxetine and thioridazine) to act as EPIs in P. mirabilis, and control crystalline biofilm formation. Both fluoxetine and thioridazine inhibited efflux in P. mirabilis, and molecular modelling predicted both drugs interact strongly with the biofilm-associated Bcr/CflA efflux system.Both EPIs were also found to significantly reduce the rate of P. mirabilis crystalline biofilm formation on catheters, and increase the time taken for catheters to block. Swimming and swarming motilies inP. mirabilis were also significantly reduced by both EPIs. The impact of these drugs on catheter biofilm formation by other uropathogens (Escherichia coli, Pseudomonas aeruginosa) was also explored, and thioridazine was shown to also inhibit biofilm formation in these species. Therefore, repurposing of existing drugs with EPI activity could be a promising approach to control catheter blockage, or biofilmformation on other medical devices

Estudios de Microbiota urinaria en personas asintomáticas y sintomáticas ¿Qué tienen los microorganismos para decir?

La vejiga urinaria se consideraba un órgano estéril, sin embargo se ha demostrado la existencia de una microbiota urinaria. Nos propusimos evaluar la presencia bacterias en la orina de personas asintomáticas y con patología u sintomáticas utilizando distintos métodos de cultivo, de bacterias intracelulares de vejiga. Las muestras se sembraron mediante urocultivo estándar -UcE- y expandido - EQUC- en distintas condiciones de cultivo y atmosféricas. La presencia de bacterias intracelulares se evaluó con una técnica de lisis celular y cultivo y mediante microscopía láser confocal (MLC). Los microorganismos presentes con diferentes morfologías en los distintos medios se identificaron por MALDI-TOF. Se recolectaron por la técnica de chorro medio 114 muestras de orina (51 de sexo femenino -7 niñas- y 63 de sexo masculino -14 niños-), de 69 asintomáticos y 45 sintomáticos. En UcE hubo crecimiento bacteriano en 91/114 muestras (79,8 %), 57 asintomáticos (83%) y 34 sintomáticos (75,6%). En EQUC, hubo crecimiento en 107/114 (94%) asintomáticos (97,1%) y 40 sintomáticos (88,9%). En asintomáticos y sintomáticos, el EQUC permite detectar crecimiento en un 15% más en relación al UcE (p=0,28 y p=0,0001). Se observaron bacterias intracelulares por la técnica de cultivo en 54/114 muestras de orina, 26/67 asintomáticos (38 %) y 28/45 sintomáticos (62 %), p=0,013. Hasta el momento se analizaron por ambas técnicas (cultivo y MLC) 87 muestras, en 38 (44%) se evidenciaron bacterias intracelulares (19/56 asintomáticos y 19/3 sintomáticos, p = 0,0023). La identificación de los microorganismos reveló u heterogeneidad de especies bacterianas presentes tanto en personas asintomáticas como sintomáticas. De las bacterias intracelulares se observó predominancia d Escherichia coli, seguido por Stenotophomonas maltophilia, Enterococcus faecalis y Proteus mirabilis. Estos estudios demuestran la presencia de bacterias en la orina de personas sanas y con sintomatología, e incluso de bacterias intracelulares. Si bien estos resultados son preliminares pueden establecer la predisposición a tener alguna patología del tracto urinario e incluso pueden sentar las bases para futuras terapias de re-establecimiento de la microbiota urinaria posterior a terapias antimicrobianas o en casos infecciones recurrentes.ANI

Development of a high-throughput ex-vivo burn wound model using porcine skin, and its application to evaluate new approaches to control wound infection

Biofilm formation in wounds is considered a major barrier to successful treatment, and has been associated with the transition of wounds to a chronic non-healing state. Here, we present a novel laboratory model of wound biofilm formation using ex-vivo porcine skin and a custom burn wound array device. The model supports high-throughput studies of biofilm formation and is compatible with a range of established methods for monitoring bacterial growth, biofilm formation, and gene expression. We demonstrate the use of this model by evaluating the potential for bacteriophage to control biofilm formation by Staphylococcus aureus, and for population density dependant expression of S. aureus virulence factors (regulated by the Accessory Gene Regulator, agr) to signal clinically relevant wound infection. Enumeration of colony forming units and metabolic activity using the XTT assay, confirmed growth of bacteria in wounds and showed a significant reduction in viable cells after phage treatment. Confocal laser scanning microscopy confirmed the growth of biofilms in wounds, and showed phage treatment could significantly reduce the formation of these communities. Evaluation of agr activity by qRT-PCR showed an increase in activity during growth in wound models for most strains. Activation of a prototype infection-responsive dressing designed to provide a visual signal of wound infection, was related to increased agr activity. In all assays, excellent reproducibility was observed between replicates using this mode

Blood Cell-Bound C4d as a Marker of Complement Activation in Patients With the Antiphospholipid Syndrome

Antiphospholipid syndrome (APS) is a chronic and disabling condition characterized by recurrent thrombosis and miscarriages mediated by antibodies against phospholipid-binding proteins (aPL), such as beta2glycoprotein I (β2GPI). Complement is involved in APS animal models and complement deposits have been documented in placenta and thrombotic vessels despite normal serum levels. Analysis of circulating blood cells coated with C4d displays higher sensitivity than the conventional assays that measure soluble native complement components and their unstable activation products in systemic lupus erythematosus (SLE). As C4d-coated blood cell count has been reported to be more sensitive than serum levels of complement components and their activation products in systemic lupus erythematosus (SLE) patients, we decided to evaluate the percentage of C4d positive B lymphocytes (BC4d), erythrocytes (EC4d), and platelets (PC4d) in primary APS patients and asymptomatic aPL positive carriers as marker of complement activation in APS. We assessed by flow cytometry the percentages of BC4d, EC4d, and PC4d in primary APS (PAPS; n. 23), 8 asymptomatic aPL positive carriers, 11 APS-associated SLE (SAPS), 17 aPL positive SLE, 16 aPL negative SLE, 8 aPL negative patients with previous thrombosis, 11 immune thrombocytopenia (ITP) patients, and 26 healthy subjects. In addition, we used an in vitro model to evaluate the ability of a monoclonal anti-β2GPI antibody (MBB2) to bind to normal resting or activated platelets and fix complement. EC4d and PC4d percentages were significantly higher in PAPS and aPL carriers as well as aPL positive SLE and SAPS than in aPL negative controls. The highest values were found in PAPS and in SAPS. The EC4d and PC4d percentages were significantly correlated with serum C3/C4 and anti-β2GPI/anti-cardiolipin IgG. In vitro studies showed that MBB2 bound to activated platelets only and induced C4d deposition. The detection of the activation product C4d on circulating erythrocytes and platelets supports the role of complement activation in APS. Complement may represent a new therapeutic target for better treatment and prevention of disability of APS patients

Tratamiento en el impuesto al valor agregado y en el impuesto a las ganancias de las mutuales de salud

Sin DatosScavone, Paola. Universidad Nacional de Rosario. Facultad de Ciencias Económicas y Estadística; Argentin

Papel de las fimbrias MR/P y flagelos de Proteus mirabilis en la colonización del tracto urinario

Orientador: Dr. Pablo Zunino.Co-orientador: Dr. Alejandro Chabalgoit

CONSERVATION AND REHABILITATION TO MUSEUM OF LAURINI PALACE IN TITO, POTENZA, ITALY

The Palace owned by the Laurini family is an important building located in the historic center of the ancient city of Tito, a few kilometers from Potenza, in the heart of Southern Italy, built between the end of the 18th century and the end of the 19th century. It was built in three phases and three factory bodies joined together at different levels (up to five), with a very irregular volume both in plan and in section and a large sloping garden with a surface of over 3,800 m2. Thanks to the attention and care of the property, the building has fully conserved its original construction, and today appears as a living manual of the architecture of its time. This also and above all because the owners refused to apply the simplistic and invasive recipes practiced after the 1980 earthquake, which led to the almost complete destruction of the historical heritage of Campania and Basilicata. The Palace preserves untidy stone wall; the vaults in stone, barrel and cross-shaped (one also lathwork type, on the top floor); the original stairs; the floors in chestnut wood, the roofs with Palladian trusses and bent tiles; the frames of doors and windows, some very ornate, and the original wooden fixtures; railings and gratins in molded cast iron and wrought iron; the pavements in colored and decorated cement grit stones; the original plasters and colorings. The Notary Laurini intends to dedicate the building to the enhancement of the thought and work of Lorenzo Ostuni, personality of Tito internationally known, artist and scholar of symbols, who died a few years ago. And to this end it has set up a Foundation to which it has given the Palace, to make it become the seat of a Museum of Symbols and related study activities. The research carried out has determined the methods through which, in respect of the historic-artistic characteristics of the building, it reaches its anti-seismic adaptation, its re-functionalization, the elimination of humidity pathologies, also achieving a marked improvement in its energy performance and sustainability

Combination of Open Subtotal Calcanectomy and Stabilization with External Fixation as Limb Salvage Procedure in Hindfoot-Infected Diabetic Foot Ulcers

Diabetic hindfoot ulcers, complicated by osteomyelitis, are associated with a high risk of major amputation. Partial calcanectomy, preceded by an effective management of the infection and of the eventual peripheral artery disease, can be considered as valid therapeutic option. We have evaluated a therapeutic protocol for diabetic hindfoot ulcers complicated by osteomyelitis, which, besides an adequate surgical debridement, considers a reconstructive pathway assisted by the positioning of a circular external fixator. We made a prospective study of a cohort of diabetic patients affected by heel ulcer complicated by osteomyelitis. All patients underwent open partial calcanectomy associated with the positioning of a circular external frame specifically designed for hindfoot stabilization and offloading. A reconstructive procedure was implemented starting with the application of negative pressure wound therapy and coverage with dermal substitute and split thickness skin grafting. From November 2014 to November 2015, 18 consecutive patients were enrolled. Mean follow-up period was 212.3 ± 64.0 days. Healing was achieved in 18 (100%) patients. The mean healing time was 69.0 ± 64.0 days. No major amputation had to be performed during the follow-up. Open partial calcanectomy associated with external fixation and skin reconstruction was as efficient as limb salvage in patients with infected lesions of the hindfoot complicated by calcaneal osteomyelitis