A new experimental model of acid- and endotoxin-induced acute lung injury in rats.

The majority of the animal models of acute lung injury (ALI) are focused on the acute phase. This limits the studies of the mechanisms involved in later phases and the effects of long-term treatments. Thus the goal of this study was to develop an experimental ALI model of aspiration pneumonia, in which diffuse alveolar damage continues for 72 h. Rats were intratracheally instilled with one dose of HCl (0.1 mol/l) followed by another instillation of one dose of LPS (0, 10, 20, 30, or 40 µg/g body weight) 2 h later, which models aspiration of gastric contents that progresses to secondary lung injury from bacteria or bacterial products. The rats were euthanized at 24, 48, and 72 h after the last instillation. The results showed that HCl and LPS at all doses caused activation of inflammatory responses, increased protein permeability and apoptosis, and induced mild hypoxemia in rat lungs at 24 h postinstillation. However, this lung damage was present at 72 h only in rats receiving HCl and LPS at the doses of 30 and 40 µg/g body wt. Mortality (∼50%) occurred in the first 48 h and only in the rats treated with HCl and LPS at the highest dose (40 µg/g body wt). In conclusion, intratracheal instillation of HCl followed by LPS at the dose of 30 µg/g body wt results in severe diffuse alveolar damage that continues at least 72 h. This rat model of aspiration pneumonia-induced ALI will be useful for testing long-term effects of new therapeutic strategies in ALI.

Innovation and technology transfer in the health sciences: a cross-sectional perspective.

This article is based on the strategic reflection and discussion that took place on occasion of the first conference on innovation and technology transfer in the health sciences organized by the REGIC-ENS-FENIN-SEMICYUC and held in Madrid in the Instituto de Salud Carlos III on May 7th, 2013, with the aim of promoting the transfer of technological innovation in medicine and health care beyond the European program "Horizon 2020". The presentations dealt with key issues such as evaluation of the use of new technologies, the need to impregnate the decisions related to adoption and innovation with the concepts of value and sustainability, and the implication of knowledge networks in the need to strengthen their influence upon the creation of a "culture of innovation" among health professionals. But above all, emphasis was placed on the latent innovation potential of hospitals, and the fact that these, being the large companies that they are, should seriously consider that much of their future sustainability may depend on proper management of their ability to generate innovation, which is not only the generation of ideas but also their transformation into products or processes that create value and economic returns.

Cloning of the sulphamidase gene and identification of mutations in Sanfilippo A syndrome.

Sanfilippo A syndrome is one of four recognised Sanfilippo sub-types (A, B, C and D) that result from deficiencies of different enzymes involved in the lysosomal degradation of heparan sulphate; patients suffer from severe neurological disorders. The Sanfilippo syndrome sub-types are also known as mucopolysaccharidosis (MPS) type III (MPS-IIIA, B, C and D), and are part of the large group of lysosomal storage disorders. Each of the MPS-III types is inherited as an autosomal recessive disorder with considerable variation in severity of clinical phenotype. The incidence of Sanfilippo syndrome has been estimated at 1:24,000 in The Netherlands with MPS IIIA (MIM #252900) the most common. MPS-IIIA is the predominant MPS-III in the United Kingdom, and has a similar high incidence to that found in The Netherlands (E. Wraith, personal communication). There is a particularly high incidence of a clinically severe form of MPS-IIIA in the Cayman Islands with a carrier frequency of 0.1 (ref. 4). Due to the mild somatic disease compared to other MPS disorders there is difficulty in diagnosing mild cases of MPS-III, hence Sanfilippo syndrome may be underdiagnosed, especially in patients with mild mental retardation. Here, we report the isolation, sequence and expression of cDNA clones encoding the enzyme sulphamidase (EC 3.10.1.1). In addition, we report the chromosomal localisation of the sulphamidase gene as being 17q25.3. An 11-bp deletion, present in sulphamidase cDNA from two unrelated Sanfilippo A patients, is described.

The future of intensive care medicine.

Intensive care medical training, whether as a primary specialty or as secondary add-on training, should include key competences to ensure a uniform standard of care, and the number of intensive care physicians needs to increase to keep pace with the growing and anticipated need. The organisation of intensive care in multiple specialty or central units is heterogeneous and evolving, but appropriate early treatment and access to a trained intensivist should be assured at all times, and intensivists should play a pivotal role in ensuring communication and high-quality care across hospital departments. Structures now exist to support clinical research in intensive care medicine, which should become part of routine patient management. However, more translational research is urgently needed to identify areas that show clinical promise and to apply research principles to the real-life clinical setting. Likewise, electronic networks can be used to share expertise and support research. Individuals, physicians and policy makers need to allow for individual choices and priorities in the management of critically ill patients while remaining within the limits of economic reality. Professional scientific societies play a pivotal role in supporting the establishment of a defined minimum level of intensive health care and in ensuring standardised levels of training and patient care by promoting interaction between physicians and policy makers. The perception of intensive care medicine among the general public could be improved by concerted efforts to increase awareness of the services provided and of the successes achieved.

[Lung-brain interaction in the mechanically ventilated patient]. / Interacción pulmón-cerebro en el paciente ventilado mecánicamente.

Patients with acute lung injury or acute respiratory distress syndrome (ARDS) admitted to the ICU present neuropsychological alterations, which in most cases extend beyond the acute phase and have an important adverse effect upon quality of life. The aim of this review is to deepen in the analysis of the complex interaction between lung and brain in critically ill patients subjected to mechanical ventilation. This update first describes the neuropsychological alterations occurring both during the acute phase of ICU stay and at discharge, followed by an analysis of lung-brain interactions during mechanical ventilation, and finally explores the etiology and mechanisms leading to the neurological disorders observed in these patients. The management of critical patients requires an integral approach focused on minimizing the deleterious effects over the short, middle or long term.

[Interpretation of ventilator curves in patients with acute respiratory failure]. / Interpretación de las curvas del respirador en pacientes con insuficiencia respiratoria aguda.

Mechanical ventilation is a therapeutic intervention involving the temporary replacement of ventilatory function with the purpose of improving symptoms in patients with acute respiratory failure. Technological advances have facilitated the development of sophisticated ventilators for viewing and recording the respiratory waveforms, which are a valuable source of information for the clinician. The correct interpretation of these curves is crucial for the correct diagnosis and early detection of anomalies, and for understanding physiological aspects related to mechanical ventilation and patient-ventilator interaction. The present study offers a guide for the interpretation of the airway pressure and flow and volume curves of the ventilator, through the analysis of different clinical scenarios.

Aggressive alveolar recruitment in ARDS: More shadows than lights.

Alveolar recruitment in acute respiratory distress syndrome (ARDS) is defined as the penetration of gas into previously unventilated areas or poorly ventilated areas. Alveolar recruitment during recruitment maneuvering (RM) depends on the duration of the maneuver, the recruitable lung tissue, and the balance between the recruitment of collapsed areas and over-insufflation of the ventilated areas. Alveolar recruitment is estimated using computed tomography of the lung and, at the patient bedside, through assessment of the recruited volume using pressure-volume curves and assessing lung morphology with pulmonary ultrasound and/or impedance tomography. The scientific evidence on RM in patients with ARDS remains subject to controversy. Randomized studies on ARDS have shown no benefit or have even reflected an increase in mortality. The routine use of RM is therefore not recommended.

Do sedation and analgesia contribute to long-term cognitive dysfunction in critical care survivors?

Deep sedation during stay in the Intensive Care Unit (ICU) may have deleterious effects upon the clinical and cognitive outcomes of critically ill patients undergoing mechanical ventilation. Over the last decade a vast body of literature has been generated regarding different sedation strategies, with the aim of reducing the levels of sedation in critically ill patients. There has also been a growing interest in acute brain dysfunction, or delirium, in the ICU. However, the effect of sedation during ICU stay upon long-term cognitive deficits in ICU survivors remains unclear. Strategies for reducing sedation levels in the ICU do not seem to be associated with worse cognitive and psychological status among ICU survivors. Sedation strategy and management efforts therefore should seek to secure the best possible state in the mechanically ventilated patient and lower the prevalence of delirium, in order to prevent long-term cognitive alterations.

Carbamazepine-loaded solid lipid nanoparticles and nanostructured lipid carriers: Physicochemical characterization and in vitro/in vivo evaluation.

Solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC) represent promising alternatives for drug delivery to the central nervous system. In the present work, four different nanoformulations of the antiepileptic drug Carbamazepine (CBZ) were designed and prepared by the homogenization/ultrasonication method, with encapsulation efficiencies ranging from 82.8 to 93.8%. The formulations remained stable at 4 °C for at least 3 months. Physicochemical and microscopic characterization were performed by photon correlation spectroscopy (PCS), transmission electron microscopy (TEM), atomic force microscopy (AFM); thermal properties by differential scanning calorimetry (DSC), thermogravimetry (TGA) and X-ray diffraction analysis (XRD). The results indicated the presence of spherical shape nanoparticles with a mean particle diameter around 160 nm in a narrow size distribution; the entrapped CBZ displayed an amorphous state. The in vitro release profile of CBZ fitted into a Baker-Lonsdale model for spherical matrices and almost the 100% of the encapsulated drug was released in a controlled manner during the first 24 h. The apparent permeability of CBZ-loaded nanoparticles through a cell monolayer model was similar to that of the free drug. In vivo experiments in a mice model of seizure suggested protection by CBZ-NLC against seizures for at least 2 h after intraperitoneal administration. The developed CBZ-loaded lipid nanocarriers displayed optimal characteristics of size, shape and drug release and possibly represent a promising tool to improve the treatment of refractory epilepsy linked to efflux transporters upregulation.

Hybrid inhalable microparticles for dual controlled release of levofloxacin and DNase: physicochemical characterization and in vivo targeted delivery to the lungs.

Current medical treatments against recurrent pulmonary infections caused by Pseudomonas aeruginosa, such as cystic fibrosis (CF) disorder, involve the administration of inhalable antibiotics. The main challenge is, however, the eradication of microbial biofilms immersed in dense mucus that requires high and recurrent antibiotic doses. Accordingly, the development of novel drug delivery systems capable of providing local and controlled drug release in the lungs is a key factor to improve the therapeutic outcome of such therapeutic molecules. Inhalable hybrid carriers were prepared by co-precipitation of CaCO3 in the presence of alginate and the resulting microparticles were treated with alginate lyase (AL) in order to modify their porosity and enhance the drug loading. The hybrid microparticles were loaded with DNase (mucolytic agent) and levofloxacin (LV, wide-spectrum antibiotic) in the range of 20-40% for LV and 28-67% for DNase, depending on the AL treatment. In vitro studies demonstrated that microparticles were able to control the DNase release for 24 h, while 30-50% of LV was released in 3 days. The morphological characterization was performed by optical, fluorescence and scanning electron microscopies, showing a narrow size distribution (5 µm). FTIR, XRD, DSC and nitrogen adsorption isotherm studies revealed the presence of the drugs in a non-crystalline state. A microcidal effect of microparticles was found on P. aeruginosa in agar plates and corroborated by Live/Dead kit and TEM observations. Finally, to study whether the microparticles improved the localization of LV in the lungs, in vivo studies were performed by pulmonary administration of microparticles to healthy mice via nebulization and dry powder inhalation, followed by the quantification of LV in lung tissue. The results showed that microparticles loaded with LV delivered the antibiotic at least 3 times more efficiently than free LV. The developed system opens the gateway to new drug delivery systems that may provide enhanced therapeutic solutions against bacterial infections and in particular as a potential tool in CF pathology.

Multicenter study of the multiple organ dysfunction syndrome in intensive care units: the usefulness of Sequential Organ Failure Assessment scores in decision making.

OBJECTIVE: This study examined the incidence and mortality of multiple organ dysfunction syndrome (MODS) in intensive care units, evaluated the limitation of life support in these patients, and determined whether daily measurement of the Sequential Organ Failure Assessment (SOFA) is useful for decision making. DESIGN AND SETTING: Prospective, observational study in 79 intensive care units. PATIENTS AND PARTICIPANTS: Of the 7,615 patients admitted during a 2-month period we found 1,340 patients to have MODS. MEASUREMENTS AND RESULTS: We recorded mortality and length of stay in the intensive care unit and the hospital and the maximum and minimum total SOFA scores during MODS. Limitation of life support in MODS patients was also evaluated. Stepwise logistic regression was used to determine the factors predicting mortality. The in-hospital mortality rate in patients with MODS was 44.6%, and some type of limitation of life support was applied in 70.6% of the patients who died. The predictive model maximizing specificity included the following variables: maximum SOFA score, minimum SOFA score, trend of the SOFA for 5 consecutive days, and age over 60 years. The model diagnostic yield was: specificity 100%, sensitivity 7.2%, positive predictive value 100%, and negative predictive value 57.3%; the area under the receiver operating characteristic curve was 0.807. CONCLUSIONS: This model showed that in our population with MODS those older than 60 years and with SOFA score higher than 9 for at least 5 days were unlikely to survive.

Structure and sequence of the human sulphamidase gene.

Sanfilippo A syndrome (MPS-IIIA) is a mucopolysaccharide lysosomal storage disorder caused by a deficiency in the lysosomal enzyme, sulphamidase (EC 3.10.1.1), which is required for the degradation of heparan sulphate. A genomic clone containing the entire sulphamidase gene was isolated from a chromosome 17-specific gridded cosmid library. The structure of the gene and the sequence of the exon/intron boundaries and the 5' promoter region were determined. The sulphamidase gene is split into 8 exons spanning approximately 11 kb.

The effect of short-term instillation of a mucolytic agent (mesna) on airway resistance in mechanically ventilated patients.

STUDY OBJECTIVE: To determine whether bolus instillation of a mucolytic agent (mesna) could diminish airway resistance, endotracheal tube resistance, or both in patients mechanically ventilated for acute respiratory failure. DESIGN: Randomized, double-blind, placebo-controlled, crossover trial. SETTING: Medical-surgical ICU of a county hospital covering 350,000 inhabitants. PATIENTS: Twenty sedated and paralyzed patients with an endotracheal tube (ET) in place more than 72 h. INTERVENTIONS: Data were recorded in three steps: (1) basal; (2) 10 min after endotracheal instillation of 3 mL of either saline solution or mesna; and (3) 10 min after instillation of the opposite drug. A 2-h washout period was allowed between data collection. MEASUREMENTS AND RESULTS: We measured tidal volume (VT), inspiratory flow (VI), auto-PEEP, peak pressure (both at airway opening [Pmax.aw] and trachea [Pmax-.tr]) and plateau pressure (Pplat), and we calculated respiratory system compliance (Crs) and the inspiratory resistances of airways+tube (Rmax.aw), airways (Rmax.tr), and ET (Rtube). We found significant differences after the instillation of mesna compared with baseline in the following: airway plus tube resistance (Rmax.aw) (16.9 +/- 7.1 vs 18.9 +/- 7.7 cm H2O); airways resistance (Rmax.tr) (9.8 +/- 6.2 vs 12.0 +/- 6.4 cm H2O), PaO2 (96 +/- 28.5 vs 80 +/- 24.8 mm Hg), PaO2/PAO2 (0.360 +/- 0.152 vs 0.296 +/- 0.127), and PaCO2 (42 +/- 12.9 vs 43 +/- 14.1 mm Hg). We found no changes in compliance, auto-PEEP, and hemodynamics during the study. Instillation of saline solution had no effect on the physiologic variables studied. CONCLUSIONS: In our patients, bolus tracheal instillation of mesna does not improve airway resistance; in fact, mesna instillation induces episodes of bronchospasm that disappear 2 h later.

Relationship between expired capnogram and respiratory system resistance in critically ill patients during total ventilatory support.

To examine the relationship of expired capnograms and respiratory system resistance (Rrs) in intubated critically ill patients, we consecutively studied 41 mechanically ventilated patients to (1) analyze the association between expired CO2 slope and auto-positive end-expiratory pressure (auto-PEEP), between Rrs and auto-PEEP, between Rrs and expired CO2 slope, and between Rrs and arterial minus end-tidal PCO2 gradient (PaCO2-PETCO2 gradient) and (2) to investigate the capacity of the expired CO2 slope and PaCO2-PETCO2 gradient to predict Rrs during mechanical ventilation. Regression analysis found a close correlation between Rrs and expired CO2 slope (r = 0.86; p < 0.001), between Rrs and auto-PEEP (r = 0.75; p < 0.001), and between auto-PEEP and expired CO2 slope (r = 0.74; p < 0.001). Weak correlation was found between Rrs and PaCO2-PETCO2 gradient (r = 0.48; p < 0.01). Prediction interval limits at 95 percent confidence level for Rrs are approximately +/- 7.39 cm H2O/L/s from the predicted value obtained by the regression equation, where Rrs = 11.42 + 2.28 expired CO2 slope. These observations suggest that CO2 elimination in critically ill patients is strongly modulated by lung, airway, endotracheal tube, and ventilator equipment resistances. Although continuous capnogram waveform monitoring at the bedside might be useful to assess Rrs, very accurate predictions could be done only in determinate patients.

Effect of PEEP on the arterial minus end-tidal carbon dioxide gradient.

The effect of PEEP on the arterial minus end-tidal carbon dioxide gradient (PaCO2-PetCO2) was evaluated in 13 adult patients with acute respiratory failure. The morphologic study of the pressure-volume (P-V) curves allowed separation of the patients into two groups: group 1 (n = 7) with initial inflection point in the (P-V) curve, and group 2 without inflection point. We hypothesized that the profile of the PaCO2-PetCO2 gradient would indicate an appropriate PEEP level only in patients with recruitable air spaces. We ventilated group 1 patients with zero end expiratory pressure (ZEEP), PEEP corresponding to inflection point pressure (PEEPPi) and PEEP5 cm H2O above PEEPPi, and group 2 patients with ZEEP, 6 cm H2O PEEP and 12 cm H2O PEEP. The PaCO2-PetCO2 gradient changed significantly in group 1 (ZEEP: 13.59 mm Hg; PEEPPi: 8.33 mm Hg; PEEPPi + 5 cm H2O: 10.54 mm Hg), but not in group 2 (ZEEP: 14.15 mm Hg; PEEP 6 cm H2O: 14.20 mm Hg; PEEP 12 cm H2O: 16.53 mm Hg). Our results show that the PaCO2-PetCO2 gradient may be useful in selecting a PEEP level which produces alveolar recruitment, but only in those patients with initial inflection point in the P-V curve.

Inflation static pressure-volume curves of the total respiratory system determined without any instrumentation other than the mechanical ventilator.

OBJECTIVE: To assess the accuracy of static pressure-volume (PV) curves of the total respiratory system performed with the data directly obtained from the Servo 900 C vs. the data obtained from an external calibrated device. DESIGN: Performance of the PV curve by the method of Levy with simultaneous recording of data obtained from both systems. SETTING: The general ICU of Hospital de Sabadell. PATIENTS: Ten sedated and paralyzed patients ventilated in the control mode for acute respiratory failure were evaluated. INTERVENTIONS AND MEASUREMENTS: Inflation static PV curves were performed by the method of Levy. We simultaneously measured airway pressure and volume by means of calibrated pressure transducer and pneumotachograph and by the internal devices built in the ventilator Siemens Servo 900 C. Statistics were concordance analysis between the two methods and covariance analysis between derived curves. RESULTS: concordance analysis between both methods showed a 95% confidence interval of (+4%, -5%) in volume and (+2.2 cmH2O, -1.7 cmH2O) in pressure. Derived PV curves analyzed by MANOVA showed no significant differences whichever the method used within subject (p = 0.579). CONCLUSION: Inflation static pressure volume curves of the total respiratory system can be accurately performed with the data directly obtained from the Servo Ventilator 900 C without the need of any other external device.