Use of the BacT/Alert MB Mycobacterial Blood Culture System for Detection of Mycobacteria in Sterile Body Fluids Other than Blood▿

The definitive diagnosis of extrapulmonary tuberculosis is made by a positive body fluid culture result. Conventional culture methods require centrifugation or filtration of body fluid (peritoneal, pleural, synovial, or pericardial fluid) in order to improve the sensitivity. The aim of the present study was to evaluate the feasibility of the direct inoculation, at the patient's bedside, of up to 5 ml of uncentrifuged fluid onto BacT/Alert MB culture bottles (bioMérieux, Durham, NC)

Identification of 54 Mycobacterial Species by PCR-Restriction Fragment Length Polymorphism Analysis of the hsp65 Gene

A total of 121 reference and clinical strains of both slowly and rapidly growing mycobacteria belonging to 54 species were studied for restriction fragment length polymorphism of a PCR-amplified 439-bp segment of the gene encoding the 65-kDa heat shock protein. Restriction digests were separated by 10% polyacrylamide gel electrophoresis (PAGE). By including a size standard in each sample, the restriction fragment profile was calculated using a computer-aided comparison program. An algorithm describing these 54 species (including 22 species not previously described) is proposed. We found that this assay based on 10% PAGE provided a more precise estimate than that based on agarose gel electrophoresis of the real size of restriction fragments as deduced from the sequence analysis and allowed identification of mycobacteria whose PCR-restriction fragment length polymorphism analysis patterns were unequivocally identified by fragments shorter than 60 bp

Mycobacterium tuberculosis in ospedale: è patogeno nosocomiale? oppure occupazionale?

La tubercolosi costituisce una emergenza sanitaria per la salute pubblica, oltre che una problematica confinata agli ambienti lavorativi. Per tale ragione le autorità sanitarie redigono ed aggiornano costantemente linee guida di notevole valore scientifico e pratico. Recenti episodi di cronaca hanno riportato alla ribalta la tubercolosi come patologia nosocomiale, la cui trasmissione può avvenire da operatore sanitario a paziente. Per quanto concerne gli operatori sanitari, una recente revisione della letteratura ha confermato che il rischio di infezione tubercolare latente e di tubercolosi sono maggiori in questa categoria rispetto alla popolazione generale. Gli strumenti di riduzione del rischio di tubercolosi in ambito nosocomiale si basano innanzitutto sull’adozione di misure di isolamento respiratorio; tuttavia tali procedure richiedono il rapido riconoscimento del sospetto clinico. La diagnosi microbiologica di tubercolosi si basa sul reperto di bacilli tubercolari nel campione clinico. La ricerca di tali microrganismi può essere eseguita con l’esame microscopico, colturale e con tecniche rapide di biologia molecolare.Tuberculosis is an emergency for public health, as well as an issue confined to working environments. For this reason, the health authorities draw up and regularly update guidelines of considerable scientific and practical value. Recent reports have shown tuberculosis as a nosocomial disease whose transmission can occur from healthcare environment to patient. With regard to healthcare workers, a recent literature review confirmed that the risk of latent TB infection and tuberculosis is greater in this category of workers than in the general population. The tools for risk reduction in nosocomial TB are based primarily on respiratory isolation measures. However such procedures require the rapid recognition of clinical suspicion. The microbiological diagnosis of tuberculosis is based on the detection of tubercular bacilli in clinical specimens. The research of these microorganisms can be done using microscopic and coltural examination as well as rapid molecular biology techniques

Impact assessment and risk analysis in the redevelopment of a healthcare structure

Any upgrading process aims for improving and optimizing operational conditions of every work environment. It generally includes the development of a change focused on organizational, technological and structural advancements in the expected conditions of specific active fields of interest. The upgrading process has to be always well planned and organized in order to be a source of growth for the structure considered and not becoming an obstacle for the daily routine of the structure itself. This aspect is much more relevant in case of health facilities since everyday activities must be constantly kept at a standard level and under an accurate control. To meet the needs of technological and legislative progress, the change following the upgrading process must be a long-term one. However, it will represent a significant variable both for the employees working conditions and the quality of the health care given to the patients. In this work an objective and complete procedure has been developed to quantify, in an impartial and univocal way, the impact that an upgrading process can have on health activities. In order to prevent and neutralize all the possible risks for patients, employees and health workers, it is of utmost importance the objectively evaluation of effects and hazards that these processes involve

Vaccine adjuvant MF59 promotes the intranodal differentiation of antigen-loaded and activated monocyte-derived dendritic cells

MF59 is an oil-in-water emulsion adjuvant approved for human influenza vaccination in European Union. The mode of action of MF59 is not fully elucidated yet, but results from several years of investigation indicate that MF59 establishes an immunocompetent environment at injection site which promotes recruitment of immune cells, including antigen presenting cells (APCs), that are facilitated to engulf antigen and transport it to draining lymph node (dLN) where the antigen is accumulated. In vitro studies showed that MF59 promotes the differentiation of monocytes to dendritic cells (Mo-DCs). Since after immunization with MF59, monocytes are rapidly recruited both at the injection site and in dLN and appear to have a morphological change toward a DC-like phenotype, we asked whether MF59 could play a role in inducing differentiation of Mo-DC in vivo. To address this question we immunized mice with the auto-fluorescent protein Phycoerythrin (PE) as model antigen, in presence or absence of MF59. We measured the APC phenotype and their antigen uptake within dLNs, the antigen distribution within the dLN compartments and the humoral response to PE. In addition, using Ovalbumin as model antigen, we measured the capacity of dLN APCs to induce antigen-specific CD4 T cell proliferation. Here, we show, for the first time, that MF59 promotes differentiation of Mo-DCs within dLNs from intranodal recruited monocytes and we suggest that this differentiation could take place in the medullary compartment of the LN. In addition we show that the Mo-DC subset represents the major source of antigen-loaded and activated APCs within the dLN when immunizing with MF59. Interestingly, this finding correlates with the enhanced triggering of antigen-specific CD4 T cell response induced by LN APCs. This study therefore demonstrates that MF59 is able to promote an immunocompetent environment also directly within the dLN, offering a novel insight on the mechanism of action of vaccine adjuvants based on emulsions

DataSheet_1_Vaccination with staphylococcal protein A protects mice against systemic complications of skin infection recurrences.docx

Skin and soft tissue infections (SSTIs) are the most common diseases caused by Staphylococcus aureus (S. aureus), which can progress to threatening conditions due to recurrences and systemic complications. Staphylococcal protein A (SpA) is an immunomodulator antigen of S. aureus, which allows bacterial evasion from the immune system by interfering with different types of immune responses to pathogen antigens. Immunization with SpA could potentially unmask the pathogen to the immune system, leading to the production of antibodies that can protect from a second encounter with S. aureus, as it occurs in skin infection recurrences. Here, we describe a study in which mice are immunized with a mutated form of SpA mixed with the Adjuvant System 01 (SpAmut/AS01) before a primary S. aureus skin infection. Although mice are not protected from the infection under these conditions, they are able to mount a broader pathogen-specific functional immune response that results in protection against systemic dissemination of bacteria following an S. aureus second infection (recurrence). We show that this “hidden effect” of SpA can be partially explained by higher functionality of induced anti-SpA antibodies, which promotes better phagocytic activity. Moreover, a broader and stronger humoral response is elicited against several S. aureus antigens that during an infection are masked by SpA activity, which could prevent S. aureus spreading from the skin through the blood.</p

A 256-Element Spiral CMUT Array with Integrated Analog Front End and Transmit Beamforming Circuits

© 2018 IEEE. MVMS-based Capacitive Micromachined Ultrasonic Transducer (CMUT) technology enables the development of advanced transducer-electronics integrated multi-chip modules (MCM) for medical imaging applications. In this pajer, we present the design, fabrication and characterization of a 256-element CMUT spiral array integrated with a 256-channel analog front end (AFE) ASIC featuring high-voltage unipolar pulsers, low-noise amplifiers and a programmable transmit beamformer. The spiral array, designed to perform volumetric beam steering at 7 MHz, is characterized by a c density-tapered layout of 220μm-wide hexagonal elements distributed over a quasi-circular area of 10 mm diameter. The CMUT and the AFE were co-designed, fabricated and interconnected through an acoustically optimized 3-D packaging method. The resulting MCM was electromechanically and acoustically characterized, demonstrating the potential of the proposed approach for integrated 2-D CMUT array fabrication.status: Published onlin

A 256-element spiral CMUT array with integrated analog front end and transmit beamforming circuits

MVMS-based Capacitive Micromachined Ultrasonic Transducer (CMUT) technology enables the development of advanced transducer-electronics integrated multi-chip modules (MCM) for medical imaging applications. In this pajer, we present the design, fabrication and characterization of a 256-element CMUT spiral array integrated with a 256-channel analog front end (AFE) ASIC featuring high-voltage unipolar pulsers, low-noise amplifiers and a programmable transmit beamformer. The spiral array, designed to perform volumetric beam steering at 7 MHz, is characterized by a c density-tapered layout of 220μm-wide hexagonal elements distributed over a quasi-circular area of 10 mm diameter. The CMUT and the AFE were co-designed, fabricated and interconnected through an acoustically optimized 3-D packaging method. The resulting MCM was electromechanically and acoustically characterized, demonstrating the potential of the proposed approach for integrated 2-D CMUT array fabrication