Clinical implications of endotoxin activity and Polymyxin-B hemoperfusion in critically ill patients with septic cardiomyopathy: A single-center, retrospective, observational study.

BACKGROUND: To explore the association between endotoxin activity (EA) and septic cardiomyopathy (SCM), the relationship between endotoxin removal by Polymyxin-B hemoperfusion (PMX-HP) and recovery from SCM (R-SCM), and the correlation between R-SCM and the 28-day mortality in septic patients admitted to the intensive care unit (ICU). METHODS: Observational study that included patients admitted to two ICUs of a tertiary university hospital between April 2011 and December 2019, who received PMX-HP for sepsis/septic shock. The SCM and R-SCM were assessed by transthoracic echocardiography. RESULTS: Among 148 patients, SCM was diagnosed in 60 (46%) of them and had no relationship with median EA (SCM group: 0.73; no-SCM group: 0.66, p = 0.48). Recovery from SCM was observed in 24 patients (49%) and was independently associated with the PMX-HP (OR 4.19, 95%CI [1.22, 14.3]; p = 0.02) and the SAPS2 II score (OR 0.94, 95%CI [0.9, 0.98]; p = 0.006). In the SCM group, the 28-day mortality was 60% and was independently predicted by R-SCM (OR 0.02, 95%CI [0.001, 0.3] p = 0.005) and SAPS II score (OR 1.11, 95%CI [1.01, 1.23] p = 0.037). CONCLUSIONS: In septic patients, EA was not associated with SCM. However, endotoxin removal by Polymyxin-B hemoperfusion was associated with recovery from cardiomyopathy, which was a predictor of lower 28-day mortality.

A case of fatal embolization during laser lithotripsy.

PURPOSE: To report in literature the first case of fatal multi-organ embolization of ureteral stones fragments during laser lithotripsy. CASE PRESENTATION: A tetraplegic 43-year-old woman was admitted to the hospital to undergo laser lithotripsy because of bilateral ureteral stones and right ureteral infected stent. During the removal of the right ureteral stent, the patient developed a sudden severe bradycardia followed by a reduction in the arterial oxygen saturation. In spite of a rapid and intensive medical intervention, the clinical picture did not improve; the woman was therefore transferred to the nearest Emergency Room where she was rescued but a cardiocirculatory arrest occurred. A claim of alleged medical malpractice was brought against the urologists. A complete autopsy was performed 8 days after death. AUTOPSY FINDINGS: The diagnosis was determined by the microscopic findings: they have unequivocally shown a massive embolization of calculus fragments in the lungs and in the heart. In the light of all these findings, the cause of death was attributable to a disseminated intravascular coagulation due to this unforeseeable embolization of calcified amorphous material. CONCLUSION: Embolization of calculus fragments represents an important challenge because it is extremely unpredictable. Indeed, a prompt diagnosis of non-thrombotic pulmonary embolism, during the urologic procedure, is extremely difficult because the condition presents with no specific clinical signs: this life-threatening pathology is often underestimated. For this reason, the autopsy and the subsequent histopathological examination are indispensable in order to prove lethal embolization: microscopic findings play a key role in the final diagnosis of death.

Characterization of multifunctional nanosystems based on the avidin-nucleic acid interaction as signal enhancers in immuno-detection.

The Avidin-Nucleic-Acids-Nano-Assembly (ANANAS) is a kind of soft poly avidin nanoparticle originating from the high affinity interaction between avidin and the nucleic acids. In this work we investigated the possibility of transforming ANANAS cores into stoichiometrically controlled multifunctional nanoparticles through a "one-pot" procedure, and we measured in a quantitative way their ability to work as reagents for enhanced immunodiagnostic detection. Initially, we measured the ANANAS loading capability for biotinylated proteins of different nature. About 200 molecules of biotin-horseradish-peroxidase (40KDa b-HRP) and 60 molecules of biotin-immunoglobulin-G (150KDa b-IgG) could be accommodated onto each nanoparticle, showing that steric limitations dictate the number of loadable entities. Stoichiometrically controlled functional assemblies were generated by mixing core particles with subsaturating amounts of b-HRP and b-IgG. When applied as detection reagents in an Enzyme-Linked-ImmunoSorbed-Assay (ELISA), these assemblies were up to two-orders of magnitude more sensitive than commercial HRP-based reagents. Assemblies of different composition displayed different efficacy, indicating that the system functionality can be fine-tuned. Within-assay variability (CV%), measured to assess if the assembly procedure is reproducible, was within 10%. Stability experiments demonstrated that the functionalyzed assemblies are stable in solution for more than one week. In principle, any biotinylated function can be loaded onto the core particle, whose high loading capacity and tunability may open the way toward further application in biomedicine.

Synthesis and chromatography-free purification of PNA-PEO conjugates for the functionalisation of gold sensors.

Peptide Nucleic Acids (PNAs) linked to high molecular weight (MW) poly(ethylene oxide) (PEO) derivatives could be useful conjugates for the direct functionalisation of gold surfaces dedicated to Surface Plasmon Resonance (SPR)-based DNA sensing. However their use is hampered by the difficulty to obtain them through a convenient and economical route. In this work we compared three synthetic strategies to obtain PNA-high MW PEO conjugates composed of (a) a 15-mer PNA sequence as the probe complementary to genomic DNA of ]Mycobacterium tuberculosis, (b) a PEO moiety (2 or 5 KDa MW) and (c) a terminal trityl-protected thiol necessary (after acidic deprotection) for grafting to gold surfaces. The 15-mer PNA was obtained by solid-phase synthesis. Its amino terminal group was later condensed to bi-functional PEO derivatives (2 and 5 KDa MW) carrying a Trt-cysteine at one end and a carboxyl group at the other end. The reaction was carried out either in solution, using HATU or PyOxim as coupling agents, or through the solid-phase approach, with 49.6%, 100% and 5.2% yield, respectively. A differential solvent extraction strategy for product purification without the need for chromatography is described. The ability of the 5 KDa PEO conjugate to function as a probe for complementary DNA detection was demonstrated using a Grating-Coupling Surface Plasmon Resonance (GC-SPR) system. The optimized PEO conjugation and purification protocols are economical and simple enough to be reproduced also within laboratories that are not highly equipped for chemical synthesis.

Effect of Fluorographene Addition on Mechanical and Adhesive Properties of a New Core Build-Up Composite.

This study aims to develop a restorative material having such mechanical and adhesive properties that it can be used both as a reconstruction material and as a luting cement. The experimental core build-up composite (CBC) was derived from a self-adhesive cement by the modification of its chemical formula, requiring the use of dedicated dentin and ceramic primers. The adhesive properties to zirconia and dentin were analyzed with a micro-Shear Bond Strength test (mSBS). The mechanical properties were analyzed by a flexural strength test. The results were compared with those obtained for other commercially available cements and core build-up materials, both before and after addition of 2 wt.% fluorographene. The CBC obtained average values in the mSBS of 49.7 ± 4.74 MPa for zirconia and 32.2 ± 4.9 MPa for dentin, as well as values of 110.9 ± 9.3 MPa for flexural strength and 6170.8 ± 703.2 MPa for Young's modulus. The addition of fluorographene, while increasing the Young's modulus of the core build-up composite by 10%, did not improve the adhesive capabilities of the primers and cement on either zirconia or dentin. The CBC showed adhesive and mechanical properties adequate both for a restoration material and a luting cement. The addition of 2 wt.% fluorographene was shown to interfere with the polymerization reaction of the material, suggesting the need for further studies.

Monitoring breathing rate by fusing the physiological impact of respiration on video-photoplethysmogram with head movements.

OBJECTIVE: The simple observation of breathing rate (BR) remains the first and often the most sensitive marker of acute respiratory dysfunction. In fact, there is evidence that drastic changes in BR are a predictive indicator of adverse events (i.e. cardiac arrest). The aim of this study is to develop a camera-based technology that may provide near-continuous estimation of BR considering the effect of respiration on video-PPG (vPPG). APPROACH: The technology has been tested in two different experimental settings, including controlled BR and more challenging scenarios with spontaneous breathing patterns. Video data were processed offline to derive the vPPG signal. The method derives respiration from beat-to-beat PPG rate and morphology changes in amplitude and width driven by the physiological relationships between vPPG and respiration. Moreover, respiratory-induced head movements were used as an additional source of information for the vPPG system. A combination of these methods has been exploited to estimate the respiratory rate every 10 seconds. MAIN RESULTS: According to the results, respiratory frequencies in the central range (0.2-0.4 Hz) may be estimated using the vPPG system with a low relative error, [Formula: see text] and interquartile range of the order [Formula: see text]. However, the vPPG system showed a drop in performance at respiratory range boundaries, around 0.1 Hz and 0.5 Hz. SIGNIFICANCE: This camera-based technology can be used as an ubiquitous BR monitoring system. However, vPPG-based systems should consider the effect of the BR in the estimation, mainly in applications where the respiratory rate is out of the 0.2-0.4 Hz range.

Gold-silver alloy semi-nanoshell arrays for label-free plasmonic biosensors.

Nanosphere lithography coupled with reactive ion etching has been used to synthesize hexagonal ordered arrays of Au-Ag bimetallic semi-nanoshells to be used as plasmonic biosensors. The degree of lateral interaction between adjacent semi-nanoshells can be controlled by tailoring the reactive ion etching time in order to boost the global plasmonic properties through the formation of near-field hot-spots, which in turn can improve the sensitivity of the biosensors. To test the efficiency of the proposed system as a biosensor, we used an established protocol for the detection of biomolecules (local sensitivity), based on the receptor-ligand approach and using the biotin-streptavidin model system. We also tested the sensitivity to a homogeneous change in the refractive index of the buffer over the sensor (bulk sensitivity). Comparing the obtained results to those of an array of nanoprisms, chosen as a benchmark, significantly higher performances both in local and bulk sensitivities have been found, in agreement with electrodynamics simulations based on finite-element methods.

Ultrasound assessment of rectus femoris and anterior tibialis muscles in young trauma patients.

PURPOSE: Quantitative and qualitative changes of skeletal muscle are typical and early findings in trauma patients, being possibly associated with functional impairment. Early assessment of muscle changes-as evaluated by muscle ultrasonography-could yield important information about patient's outcome. METHODS: In this prospective observational study, we used ultrasonography to evaluate the morphological changes of rectus femoris (RF) and anterior tibialis (AT) muscles in a group of young, previously healthy trauma patients on enteral feeding. RESULTS: We studied 38 severely injured patients (median Injury Severity Score = 34; median age = 40 y.o.) over the course of the ICU stay up to 3 weeks after trauma. We found a progressive loss of muscle mass from day 0 to day 20, that was more relevant for the RF (45%) than for the AT (22%); this was accompanied by an increase in echogenicity (up to 2.5 by the Heckmatt Scale, where normal echogenicity = 1), which is an indicator of myofibers depletion. CONCLUSIONS: Ultrasound evaluation of skeletal muscles is inexpensive, noninvasive, simple and easily repeatable. By this method, we were able to quantify the morphological changes of skeletal muscle in trauma patients. Further studies may rely on this technicque to evaluate the impact of different therapeutic strategies on muscle wasting.

Supported imprinted nanospheres for the selective recognition of cholesterol.

The preparation of innovative polymeric systems using molecular imprinting technology for application in extracorporeal blood purification is described. Membranes based on a methylmethacrylate-co-acrylic acid copolymer, produced through the phase inversion method, were modified introducing into their structure specific binding sites for cholesterol molecule by adding molecularly imprinted nanoparticles in the membrane matrix. Membranes prepared are intended to selectively remove cholesterol from the blood by using interactions at a molecular level, between the membrane/nanoparticles devices and the template, created during the preparation of polymers. Three polymeric systems in form of nanoparticles were prepared differing in the polymerisation solvent (a mixture of acetonitrile and ethanol (1:1) or pure ethanol), and the molar ratio between the functional monomer and the cross-linker (2.3:1 and 1:1). Two out of three of the prepared polymers showed a very good template rebinding capacity both in phosphate buffer solution (pH 6.9) and in ethanol. In particular the nanoparticles rebound 115.4 mg cholesterol/g polymer in buffer solution, and 57 mg cholesterol/g polymer in ethanol. The deposition of the nanoparticles on the surface of the phase inversion membranes produced devices with interesting rebinding performances towards cholesterol in buffer solution: a specific recognition of 14.09 mg cholesterol/g system (membrane and nanoparticles) was detected, indicating maintained binding capacity of supported particles as well.

Enzymatic erosion of bioartificial membranes to control drug delivery.

The preparation of an enzymatic controlled drug release system from blends of PVA/starch/alphaA, in the form of films, is described. It was shown that alphaA hydrolyses the starch within these films, resulting in a time-dependent change of the porosity in the matrix. Films were characterized by calorimetric analysis to study the interactions between the enzyme and the polymeric constituents at the molecular level. The presence of alphaA, in fact, influenced the PVA crystallization in the blends. Release tests and permeability experiments were carried out to evaluate the transport properties of the films. An increase in porosity and permeability was observed by increasing alphaA content (16-28 wt.-%). Films loaded with theophylline and caffeine were also prepared to analyze drug release properties of the matrix. Drug release kinetics were coherent with the measured changes in porosity: at higher alphaA concentrations the amount of released drug increased under the influence of diffusion and erosion processes. The results obtained are promising for the realization of drug delivery devices for a rapid release or for the release of poorly soluble drugs which usually remain entrapped in the matrix.SEM images of a PVA/starch/alphaA film before (A) and after (B) the erosion.

Immunonutrients in critically ill patients: an analysis of the most recent literature.

Modulation of inflammatory and immune response to critical illness has been the goal of much research in the last decade and a variety of drugs and nutrients (so called "immunonutrients") have been tested in experimental models with promising results. Though, in the clinical setting of intensive care, their efficacy have been inconsistently proven, most likely because the effects of each drug may vary in relation to the timing, the dose, the route of administration, the interaction with other nutrients, the severity of illness and many other factors. Though the early studies of the beginning of this century (2000-2009) have shown some clinical benefits, recent multicenter trials (2011-2015) have failed to prove a consistent benefit of immunonutrition in terms of mortality or other clinical endpoints. Reviewing the latest evidence-based documents on this subject (multicenter trials, systematic reviews, meta-analyses and international guidelines), there is no convincing evidence that immunonutrients may be beneficial in the critically ill. Considering that these substances invariably increase the costs of health care and may be unsafe or even harmful in some subgroups, particularly in septic patients, we conclude that routine administration of immune-nutrients (glutamine, arginine, omega-3 fatty acids, selenium, etc.) cannot be currently recommended in the critically ill.

In vivo fate of avidin-nucleic acid nanoassemblies as multifunctional diagnostic tools.

This study describes the formulation optimization and body-cell distribution and clearance in mice of a dually fluorescent biodegradable poly avidin nanoassembly based on the novel Avidin-Nucleic-Acid-Nano-ASsembly (ANANAS) platform as a potential advancement of classic avidin/biotin-based targeted delivery. The nanoformulation circulates freely in the bloodstream; it is slowly captured by filter organs; it is efficiently cleared within 24-48 h, and it is poorly immunogenic. The system displays more favorable properties than its parent monomeric avidin and it is a promising tool for diagnostic purposes for future translational aims, for which free circulation in the bloodstream, safety, multifunctionality and high composition definition are all necessary requirements. In addition, the assembly shows a time-dependent cell penetration capability, suggesting it may also function as a NP-dependent drug delivery tool. The ease of preparation together with the possibility to fine-tune the surface composition makes it also an ideal candidate to understand if and how nanoparticle composition affects its localization.

Integrated architecture for the electrical detection of plasmonic resonances based on high electron mobility photo-transistors.

We report the design of an integrated platform for on-chip electrical transduction of the surface plasmon resonance supported by a nanostructured metal grating. The latter is fabricated on the active area of a GaAs/AlGaAs photo-HEMT and simultaneously works as the electronic gate of the device. The gold plasmonic crystal has a V-groove profile and has been designed by numerical optical simulations. By showing that the numerical models accurately reproduce the phototransistors experimental response, we demonstrate that the proposed architecture is suitable for the development of a new class of compact and scalable SPR sensors.

Synthesis and characterization of copolymers of methylmethacrylate and 2-hydroxyethyl methacrylate for the aqueous solubilization of Paclitaxel.

The aim of the present work is the modification of a hydrophobic polymeric macromolecule, polymethylmethacrylate, by introducing hydrophilic moieties of 2-hydroxyethyl methacrylate within the polymer chain. Synthesis, characterization, and drug delivery control capabilities exerted on a highly hydrophobic drug (Paclitaxel) are illustrated. In particular, the dependency of the drug delivery kinetic on the fraction of hydrophilic units inserted in the copolymer chain was studied. Results showed that it is possible to have an increase of the kinetic delivery introducing hydrophilic units. In addition, a double control, diffusive and due to the relaxation of the molecules, on drug delivery was obtained.