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INTRODUCTION: Pompe disease is caused by a rare biallelic mutation in the GAA gene resulting in acid α-glucosidase deficiency and glycogen accumulation. AIM: We analyzed hospital admissions associated with the administration of Myozyme®, utilizing the French hospital discharge database, known in France as the Programme de Médicalisation des Systèmes d'Information (PMSI), which comprehensively captures all hospital activity within the country. METHODS: In this observational study, we examined hospitalization records from April 4, 2012, to December 31, 2019, within the PMSI database, focusing on admissions where Myozyme® was administered. We particularly investigated the incidence of critical care admissions and adverse events (AEs) related to Myozyme®. RESULTS: From 2012 to 2019, approximately 26,714 hospital stays involving Myozyme® administration were recorded for 239 patients. Most (96.6%) of these were outpatient stays, with only 3.2% in critical care. Furthermore, hospitalizations without critical care needs increased from 96% in 2012 to 99% in 2019. Of the patients receiving at least one infusion, 997 critical care admissions were recorded, with 781 (78.3%) occurring concurrent with or the day after the Myozyme® treatment without directly correlating to adverse effects of enzyme therapy. CONCLUSIONS: The analysis of the French hospital discharge database indicated that Myozyme® was associated with a low incidence of AEs and complications in a hospital context, supporting the consideration of its safe use in home-infusion settings.

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Metal-organic frameworks (MOFs), a diverse and rapidly expanding class of crystalline materials, present many opportunities for various applications. Within this class, the amino-functionalized Zr-MOF, namely, UiO-66-NH2, stands out due to its distinctive chemical and physical properties. In this study, we report on the new unique property where UiO-66-NH2 nanocrystals exhibited enhanced fluorescence upon heating, which was persistently maintained postcooling. To unravel the mechanism, the changes in the fluorescence signal were monitored by steady-state fluorescence spectroscopy, lifetime measurements, and a fluorescence microscope, which revealed that upon heating, multiple mechanisms could be contributing to the observed enhancement; the MOFs can undergo disaggregation, resulting in a fluorescent enhancement of the colloidally stable MOF nanocrystals and/or surface-induced phenomena that result in further fluorescence enhancement. This observed temperature-dependent photophysical behavior has substantial applications. It not only provides pathways for innovations in thermally modulated photonic applications but also underscores the need for a better understanding of the interactions between MOF crystals and their environments.

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Background: Current guidelines recommend intramuscular botulinum toxin type A (BoNT-A) injection as first-line treatment for spasticity, a frequent and impairing feature of various central nervous system (CNS) lesions such as stroke. Patients with spasticity commonly require BoNT-A injections once every 3 to 4 months. We conducted a nationwide, population-based, retrospective cohort study, using the French National Hospital Discharge Database (PMSI), to describe BoNT-A use for spasticity in clinical practice in France between 2014 and 2020. The PMSI database covers the whole French population, corresponding to over 66 million persons. Methods: We first searched the PMSI database for healthcare facility discharge of patients who received BoNT-A injections between 2014 and 2020, corresponding to the first set. For each BoNT-A-treated patient, we identified the medical condition for which BoNT-A may have been indicated. Another search of the PMSI database focused on patients admitted for acute stroke between 2014 and 2016 and their spasticity-related care pathway (second set). Overall, two subpopulations were analysed: 138,481 patients who received BoNT-A injections between 2014 and 2020, and 318,025 patients who survived a stroke event between 2014 and 2016 and were followed up until 2020. Results: Among the 138,481 BoNT-A-treated patients, 53.5% received only one or two BoNT-A injections. Most of these patients (N = 85,900; 62.0%) received BoNT-A because they had CNS lesions. The number of patients with CNS lesions who received ≥1 BoNT-A injection increased by a mean of 7.5% per year from 2014 to 2019, but decreased by 0.2% between 2019 and 2020, corresponding to the COVID-19 outbreak. In stroke survivors (N = 318,025), 10.7% were coded with post-stroke spasticity, 2.3% received ≥1 BoNT-A injection between 2014 and 2020, and only 0.8% received ≥3 injections within the 12 months following BoNT-A treatment initiation, i.e., once every 3 to 4 months. Conclusion: Our analysis of the exhaustive PMSI database showed a suboptimal implementation of BoNT-A treatment recommendations in France. BoNT-A treatment initiation and re-administration are low, particularly in patients with post-stroke spasticity. Further investigations may help explain this observation, and may target specific actions to improve spasticity-related care pathway.

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In this work, we utilized electrospinning to develop advanced composite membranes of polyvinyl chloride (PVC) loaded with postmetalated metal-organic frameworks (MOFs), specifically UiO-66(COOH)2-Ag and ZIF-8-Ag. This innovative technique led to the creation of highly stable PVC/MOFs-Ag membrane composites, which were thoroughly characterized using various analytical techniques, including scanning electron microscopy, powder X-ray diffraction, thermogravimetric analysis, X-ray photoelectron spectroscopy, porosity analysis, and water contact angle measurement. The results verified the successful integration of MOF crystals within the nanofibrous PVC membranes. The obtained composites exhibited larger fiber diameters for 5 and 10% MOF loadings and a smaller diameter for 20% loading. Additionally, they displayed greater average pore sizes than traditional PVC membranes across most MOF loading percentages. Furthermore, we examined the antibacterial properties of the fabricated membranes at different MOFs-Ag loadings. The findings revealed that the membranes demonstrated significant antibacterial activity up to 95% against both Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria as the MOFs-Ag loading increased, even when maintaining a constant silver concentration. This indicates a contact-based inhibition mechanism. The outcomes of this study have crucial implications for the development of novel, stable, and highly effective antibacterial materials, which could serve as superior alternatives for face masks and be integrated into materials requiring regular decontamination, as well as potential water filtration systems.

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There are limited real-world data on the use of botulinum toxin type A (BoNT-A) in patients with multiple sclerosis (MS). Accordingly, this nationwide, population-based, retrospective cohort study aimed to describe BoNT-A treatment trends in patients with MS between 2014 and 2020 in France. This study extracted data from the French National Hospital Discharge Database (Programme de Médicalisation des Systèmes d'Information, PMSI) covering the entire French population. Among 105,206 patients coded with MS, we identified those who received ≥1 BoNT-A injection, administered within striated muscle for MS-related spasticity and/or within the detrusor smooth muscle for neurogenic detrusor overactivity (NDO). A total of 8427 patients (8.0%) received BoNT-A injections for spasticity, 52.9% of whom received ≥3 BoNT-A injections with 61.9% of the repeated injections administered every 3 to 6 months. A total of 2912 patients (2.8%) received BoNT-A injections for NDO, with a mean of 4.7 injections per patient. Most repeated BoNT-A injections within the detrusor smooth muscle (60.0%) were administered every 5 to 8 months. There were 585 patients (0.6%) who received both BoNT-A injections within striated muscle and the detrusor smooth muscle. Overall, our study highlights a broad range of BoNT-A treatment practices between 2014 and 2020 in patients with MS.

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Thermal mapping in biological membranes could unlock and help us understand many chemical and physical processes that do not only pertain to localized membrane phenomena but also extend to many other intra- and extracellular pathways. In this manuscript, we report the development of a ratiometric thermal fluorescent probe based on the Förster resonance energy transfer between a lipid-embedded conjugated polyelectrolyte and a lyophilic acceptor dye. We showed that the Förster resonance energy transfer (FRET) pair is sensitive within the relevant physiological temperature window (20.0-50.0 °C). The signal was also shielded from an external pH and stable when cycled multiple times. The probe was also sensitive to the membrane composition and could, therefore, be further developed to probe the membrane composition and viscosity.

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OBJECTIVE: The goal of this study was to assess the differences between an ex ante and an ex post cost-effectiveness analysis of Dabigatran etexilate vs VKAs for the prevention of thromboembolic events in non-valvular atrial fibrillation patients and to draw lessons on the design and use of real-world data for decision making. METHODS: The same model was used to calculate the cost-effectiveness ratio using two sets of parameters. One set included the efficacy and safety outcomes data from RE-LY, the pivotal trial comparing Dabigatran to warfarin; cost data came from an ex ante publication. Outcomes data for the second set came from real-world data studies. Cost data were a mix of real-world data and other sources. Two treatment strategies were compared: treatment initiation by either Dabigatran or VKAs, followed by either VKAs or Dabigatran. A crude comparison of results was performed; the impact of data differences was then assessed. Probabilistic sensitivity results of the two analyses were compared. RESULTS: With real-world evidence, Dabigatran at both dosages was more effective for the prevention of ischemic strokes, intra-cranial haemorrhages, with less major extra-cranial haemorrhages and a similar risk of myocardial infarction. Using clinical trial data, Dabigatran150 mg (resp. Dabigatran110 mg) as a first-line treatment vs VKAs yielded an ICER of € 8077/QALY (resp. € 13,116/QALY). Real-world evidence scenarios were cost-saving and more effective for both dosages. CONCLUSION: The reassessment of outcomes and cost data had an impact on results, improving the efficiency of Dabigatran. We identify methodological issues which should be discussed if post-launch RWE based cost-effectiveness data become a standard in HTA decision making.

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Thermal sensing in thin films polymers has been a significant limitation towards optimizing the heat dissipation in micro- and nano-electronic devices as well as many other thin film-based technologies. In this work, we report on poly (phenylene ethynylene) fluorescent-based conjugated polyelectrolyte capable of detecting thermal fluctuations in polymer films prepared from polyvinylpyrrolidone-co-vinyl acetate. The sensor was first optimized in solution by testing two polyvinylpyrrolidone (PVP) copolymers (co-vinyl acetate (VA) and co-polystyrene (PS)) before it was spun cast onto quartz slides and imaged using a DSLR camera at different temperatures. The images were analyzed and showed a change in color with the increase in temperature. When not illuminated, the polymer thin film is clear and transparent.

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Porphyrins have been widely used in many optical devices given their unique photochemical properties. Their poor photostability has, however, limited their wide applications in bioimaging and biosensing schemes. Herein, we report the remarkable photostability enhancement of the porphyrin, carboxyphenyl porphyrin (TCPP-H2) when locked in a zirconium based metal-organic framework (MOF-525). Steady-state ensemble fluorescence spectroscopy experiments showed minimal changes (2%) in the recorded signal when MOF-525 was continuously illuminated as compared to a 16% decrease for free porphyrins. Single particle fluorescence imaging revealed bright microparticles with exceptional photostability and no-blinking within the experiment window. This study highlights the use of metal-organic frameworks for preparing photostable microstructures by leveraging on their unique self-assembly properties.

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It is generally accepted that positively charged molecules are the gold standard to by-pass the negatively charged cell membrane. Here, it is shown that cellular uptake is also possible for polymers with negatively charged side chains and hydrophobic backbones. Specifically, poly[5-methoxy-2-(3-sulfopropoxy)-1,4-phenylenevinylene], a conjugated polyelectrolyte with sulfonate, as water-soluble functional groups, is shown to accumulate in the intracellular region. When the polymer hydrophobic backbone is dissolved using polyvinylpyrrolidone, an amphiphilic macromolecule, the cellular uptake is dramatically reduced. The report sheds light on the fine balance between negatively charged side groups and the hydrophobicity of polymers to either enhance or reduce cellular uptake. As a result, these findings will have important ramifications on the future design of targeted cellular delivery nanocarriers for imaging and therapeutic applications.

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There is an increasing need to develop new antibacterial materials for treating contaminated potable water, particularly for addressing increasing bacterial resistance, which is predicted to be a major threat to human health in the future, especially for high-risk groups and populations displaced in conflict regions. In addition, a material that can be easily separated from solutions is highly desirable to allow for the fast and efficient recovery of the bactericidal material. In this study, we reported the preparation of microsized cross-linked PVP (PVPP) decorated with silver nanoparticles. The silver particles grew to sizes between 100 and 700 nm with average diameters of 534 ± 72 nm (N = 70), 265 ± 66 nm (N = 70), and 255 ± 75 nm (N = 70) for the 0.5, 1, and 10 mg/mL concentrations of PVPP, respectively. The prepared complexes were easily separated from water by gravity. The minimum inhibitory concentrations against E. coli were calculated to be approximately 100, 65, and 60 µg/mL Ag+ for the 10, 1, and 0.5 mg/mL concentrations of PVPP, respectively. The PVPP/AgNP microparticles retained their antibacterial activity after they were collected from a bacterial culture, washed, and then recycled into a second batch of freshly prepared microorganism solution.

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Metal-organic frameworks (MOFs) have emerged as an important class of hybrid organic-inorganic materials. One of the reasons they have gained remarkable attention is attributed to the possibility of altering them by postsynthetic modification, thereby providing access to new and novel advanced materials. MOFs have been applied in catalysis, gas storage, gas separation, chemical sensing, and drug delivery. However, their bactericidal use has rarely been explored. Herein, we developed a two-step process for the synthesis of zirconium-based MOFs metalated with silver cations as a potent antibacterial agent. The obtained products were thoroughly characterized by powder X-ray diffraction, scanning electron microscopy, UV-visible, IR, thermogravimetric, and Brunauer-Emmett-Teller analyses. Their potency was evaluated against E. coli with a reported minimal inhibitory concentration and minimal bactericidal concentration of as low as 6.5 µg/mL of silver content. Besides the novelty of the system, the advantage of this strategy is that the MOFs could be potentially regenerated and remetalated after each antibacterial test, unlike previously reported frameworks, which involved the destruction of the framework.

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We report a simple, generally applicable, and noninvasive fluorescent method for mapping thermal fluctuations in hydrogel matrices using an unmodified commercially available digital single-lens reflex camera (DSLR). The nanothermometer is based on the complexation of short conjugated polyelectrolytes, poly(phenylene ethynylene) carboxylate, with an amphiphilic polymer, polyvinylpyrrolidone, which is in turn trapped within the porous network of a gel matrix. Changes in the temperature lead to a fluorescent ratiometric response with a maximum relative sensitivity of 2.0% and 1.9% at 45.0 °C for 0.5% agarose and agar, respectively. The response was reversible with no observed hysteresis when samples were cycled between 20 and 40 °C. As a proof of concept, the change in fluorescent signal/color was captured using a digital camera. The images were then dissected into their red-green-blue (RGB) components using a Matlab routine. A linear correlation was observed between the hydrogel temperature and the green and blue intensity channels. The reported sensor has the potential to provide a wealth of information when thermal fluctuations mapped in soft gels matrices are correlated with chemical or physical processes.

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We report a self-referenced ratiometric nanothermometer based on short conjugated polyelectrolytes. An amphiphilic macromolecule destabilizes the polymer π-π stacking and makes it possible to shift the equilibrium between the less emissive aggregated state (520 nm) and the brighter individual chain (450 nm) within 20.0 °C and 70.0 °C.

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We present a general and straightforward one-step approach to enhance the photophysical properties of conjugated polyelectrolytes. Upon complexation with an amphiphilic polymer (polyvinylpyrrolidone), an anionic conjugated polyelectrolyte (poly[5-methoxy-2-(3-sulfopropoxy)-1,4-phenylenevinylene]) was prepared into small nanoparticles with exceptional photostability and brightness. The polymer fluorescence intensity was enhanced by 23 -fold and could be easily tuned by changing the order of addition. Single molecule experiments revealed a complete suppression of blinking. In addition, after only losing 18% of the original intensity, a remarkable amount of photons were emitted per particle (∼10(9), on average). This number is many folds greater than popular organic fluorescent dyes. We believe that an intimate contact between the two polymers is shielding the conjugated polyelectrolyte from the destructive photooxidation. The prepared nanohybrid particles will prove instrumental in single particle based fluorescent assays and can serve as a probe for the current state-of-the-art bioimaging fluorescence techniques.

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Here we report spectroscopic studies on the interaction of negatively charged, amphiphilic polyphenylene ethynylene (PPE) polymers with liposomes prepared either from negative, positive or zwitterionic lipids. Emission spectra of PPEs of 7 and 49 average repeat units bearing carboxylate terminated side chains showed that the polymer embeds within positively charged lipids where it exists as free chains. No interaction was observed between PPEs and negatively charged lipids. Here the polymer remained aggregated giving rise to broad emission spectra characteristic of the aggregate species. In zwitterionic lipids, we observed that the majority of the polymer remained aggregated yet a small fraction readily embedded within the membrane. Titration experiments revealed that saturation of zwitterionic lipids with polymer typically occurred at a polymer repeat unit to lipid mole ratio close to 0.05. No further membrane embedding was observed above that point. For liposomes prepared from positively charged lipids, saturation was observed at a PPE repeat unit to lipid mole ratio of ∼0.1 and liposome precipitation was observed above this point. FRET studies showed that precipitation was preceded by lipid mixing and liposome fusion induced by the PPEs. This behavior was prominent for the longer polymer and negligible for the shorter polymer at a repeat unit to lipid mole ratio of 0.05. We postulate that fusion is the consequence of membrane destabilization whereby the longer polymer gives rise to more extensive membrane deformation than the shorter polymer.

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The hepatitis C virus (HCV) non-structural protein 5B (NS5B) is an RNA-dependent RNA polymerase that is essentially required for viral replication. Although previous studies revealed important properties of static NS5B-RNA complexes, the nature and relevance of dynamic interactions have yet to be elucidated. Here, we devised a single molecule Förster Resonance Energy Transfer (SM-FRET) assay to monitor temporal changes upon binding of NS5B to surface immobilized RNA templates. The data show enzyme association-dissociation events that occur within the time resolution of our setup as well as FRET-fluctuations in association with stable binary complexes that extend over prolonged periods of time. Fluctuations are shown to be dependent on the length of the RNA substrate, and enzyme concentration. Mutations in close proximity to the template entrance (K98E, K100E), and in the center of the RNA binding channel (R394E), reduce both the population of RNA-bound enzyme and the fluctuations associated to the binary complex. Similar observations are reported with an allosteric nonnucleoside NS5B inhibitor. Our assay enables for the first time the visualization of association-dissociation events of HCV-NS5B with RNA, and also the direct monitoring of the interaction between HCV NS5B, its RNA template, and finger loop inhibitors. We observe both a remarkably low dissociation rate for wild type HCV NS5B, and a highly dynamic enzyme-RNA binary complex. These results provide a plausible mechanism for formation of a productive binary NS5B-RNA complex, here NS5B slides along the RNA template facilitating positioning of its 3' terminus at the enzyme active site.

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The post-genomics era has brought about new Omics biotechnologies, such as proteomics and metabolomics, as well as their novel applications to personal genomics and the quantified self. These advances are now also catalyzing other and newer post-genomics innovations, leading to convergences between Omics and nanotechnology. In this work, we systematically contextualize and exemplify an emerging strand of post-genomics life sciences, namely, nanoproteomics and its applications in health and integrative biological systems. Nanotechnology has been utilized as a complementary component to revolutionize proteomics through different kinds of nanotechnology applications, including nanoporous structures, functionalized nanoparticles, quantum dots, and polymeric nanostructures. Those applications, though still in their infancy, have led to several highly sensitive diagnostics and new methods of drug delivery and targeted therapy for clinical use. The present article differs from previous analyses of nanoproteomics in that it offers an in-depth and comparative evaluation of the attendant biotechnology portfolio and their applications as seen through the lens of post-genomics life sciences and biomedicine. These include: (1) immunosensors for inflammatory, pathogenic, and autoimmune markers for infectious and autoimmune diseases, (2) amplified immunoassays for detection of cancer biomarkers, and (3) methods for targeted therapy and automatically adjusted drug delivery such as in experimental stroke and brain injury studies. As nanoproteomics becomes available both to the clinician at the bedside and the citizens who are increasingly interested in access to novel post-genomics diagnostics through initiatives such as the quantified self, we anticipate further breakthroughs in personalized and targeted medicine.

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Understanding how living cells interact with nanostructures is integral to a better understanding of the fundamental principles of biology and the development of next-generation biomedical/bioenergy devices. Recent studies have demonstrated that mammalian cells can recognize nanoscale topographies and respond to these structures. From this perspective, there is a growing recognition that nanostructures, along with their specific physicochemical properties, can also be used to regulate the responses and motions of bacterial cells. Here, by utilizing a well-defined silicon nanowire array platform and single-cell imaging, we present direct evidence that Shewanella oneidensis MR-1 can recognize nanoscale structures and that their swimming patterns and initial attachment locations are strongly influenced by the presence of nanowires on a surface. Analyses of bacterial trajectories revealed that MR-1 cells exhibited a confined diffusion mode in the presence of nanowires and showed preferential attachment to the nanowires, whereas a superdiffusion mode was observed in the absence of nanowires. These results demonstrate that nanoscale topography can affect bacterial movement and attachment and play an important role during the early stages of biofilm formation.

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The generation of site-specific bioconjugates of proteins is highly desired for a number of biophysical and nanotechnological applications. To this end, many strategies have been developed that allow the specific modification of certain canonical amino acids and, more recently, noncanonical functional groups. P450 enzymes are heme-dependent monooxygenases involved in xenobiotic metabolism and in the biosynthesis of a variety of secondary metabolites. We became interested in the site-specific modification of these enzymes, CYP3A4 in particular, through our studies of their in vitro biocatalytic properties and our desire to exploit their remarkable ability to oxidize unactivated C-H bonds in a regio- and stereospecific manner. Obtained via a partial cysteine-depletion approach, a functional triple mutant of CYP3A4 (C98S/C239S/C468G) is reported here which is singly modified at C64 by maleimide-containing groups. While cysteine-labeling of the wild-type enzyme abolished >90% of its enzymatic activity, this mutant retained ≥75% of the activity of the unmodified wild-type enzyme with 9 of the 18 maleimides that were tested. These included both fluorescent and solid-supported maleimides. The loss of activity observed after labeling with some maleimides is attributed to direct enzyme inhibition rather than to steric effects. We also demonstrate the functional immobilization of this mutant on maleimide-functionalized agarose resin and silica microspheres.

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