Measurements of Na+-occluded intermediates during the catalytic cycle of the Na+/K+-ATPase provide novel insights into the mechanism of Na+ transport.

The Na+/K+-ATPase is an integral plasma membrane glycoprotein of all animal cells that couples the exchange of intracellular Na+ for extracellular K+ to the hydrolysis of ATP. The asymmetric distribution of Na+ and K+ is essential for cellular life and constitutes the physical basis of a series of fundamental biological phenomena. The pumping mechanism is explained by the Albers-Post model. It involves the presence of gates alternatively exposing Na+/K+-ATPase transport sites to the intracellular and extracellular sides and includes occluded states in which both gates are simultaneously closed. Unlike for K+, information is lacking about Na+-occluded intermediates, as occluded Na+ was only detected in states incapable of performing a catalytic cycle, including two Na+-containing crystallographic structures. The current knowledge is that intracellular Na+ must bind to the transport sites and become occluded upon phosphorylation by ATP to be transported to the extracellular medium. Here, taking advantage of epigallocatechin-3-gallate to instantaneously stabilize native Na+-occluded intermediates, we isolated species with tightly bound Na+ in an enzyme able to perform a catalytic cycle, consistent with a genuine occluded state. We found that Na+ becomes spontaneously occluded in the E1 dephosphorylated form of the Na+/K+-ATPase, exhibiting positive interactions between binding sites. In fact, the addition of ATP does not produce an increase in Na+ occlusion as it would have been expected; on the contrary, occluded Na+ transiently decreases, whereas ATP lasts. These results reveal new properties of E1 intermediates of the Albers-Post model for explaining the Na+ transport pathway.

Bio-based polyester-polyurethane foams: synthesis and degradability by Aspergillus niger and Aspergillus clavatus.

In this article, the degradability by Aspergillus niger and Aspergillus clavatus of three bio-based polyurethane (PU) foams is compared to previous degradability studies involving a Pseudomonas sp. bacterium and similar initial materials (Spontón et al. in Int. Biodet. Biodeg. 85:85-94, 2013, https://doi.org/10.1016/j.ibiod.2013.05.019 ). First, three new polyester-polyurethane foams were prepared from mixtures of castor oil (CO), maleated castor oil (MACO), toluene diisocyanate (TDI), and water. Then, their degradation tests were carried out in an aqueous medium, and employing the two mentioned fungi, after their isolation from the environment. From the degradation tests, the following was observed: (a) the insoluble (and slightly collapsed) foams exhibited free hydroxyl, carboxyl, and amine moieties; and (b) the water soluble (and low molar mass) compounds contained amines, carboxylic acids, and glycerol. The most degraded foam contained the highest amount of MACO, and therefore the highest concentration of hydrolytic bonds. A basic biodegradation mechanism was proposed that involves hydrolysis and oxidation reactions.

Ultrasound-guided caudal quadratus lumborum block combined with the greater ischiatic notch plane block as motor-protective analgesia for the pelvic limb in dogs.

OBJECTIVE: To develop an ultrasound-guided caudal quadratus lumborum block (C-QLB) technique in canine cadavers and to compare sensory and motor blockade resulting from the combination of ultrasound-guided greater ischiatic notch (GIN) plane and C-QLB approaches (GIN-CQLB group) versus a lumbosacral plexus (LSP group) approach [combination of lateral pre-iliac (LPI) and parasacral (PS) techniques] in dogs. STUDY DESIGN: Descriptive anatomical study and prospective randomized, blinded, experimental crossover trial. ANIMALS: A total of six canine cadavers and six adult Beagle dogs. METHODS: Phase I: following ultrasound-guided C-QLB injections of 0.3 mL kg-1 of dye, using the interfascial plane located lateral to the quadratus lumborum muscle at the level of the sixth lumbar vertebra (L6) as injection point, the spread of injectate and nerve staining was evaluated using gross anatomical dissection. PHASE II: sensory and motor blockade achieved with the GIN-CQLB or LSP blocks in Beagle dogs were evaluated and compared. The assigned technique was performed with 2% lidocaine: 0.2 mL kg-1 for the GIN and PS approaches and 0.3 mL kg-1 for the C-QLB and LPI approaches. RESULTS: Dissection revealed distribution of dye around the lumbar hypaxial musculature, extending into the paravertebral spaces, with staining of 3 (2-4) [median (interquartile range)] spinal nerves, spanning L3 to L6. The median motor blockade in the GIN-CQLB and LSP groups was 7 (7-8) versus 16 (10-16) (p = 0.026), whereas the median sensory blockade was 5 (4-5) versus 3 (3-3) (p = 0.025), respectively. CONCLUSION AND CLINICAL SIGNIFICANCE: The GIN-CQLB approach desensitized the thigh dermatomes effectively. Compared with the LSP approaches, GIN-CQLB exhibits a motor-protective effect by preserving tonic muscle function.

Polyelectrolyte-multivalent molecule complexes: physicochemical properties and applications.

The complexation of polyelectrolytes with other oppositely charged structures gives rise to a great variety of functional materials with potential applications in a wide spectrum of technological fields. Depending on the assembly conditions, polyelectrolyte complexes can acquire different macroscopic configurations such as dense precipitates, nanosized colloids and liquid coacervates. In the past 50 years, much progress has been achieved to understand the principles behind the phase separation induced by the interaction of two oppositely charged polyelectrolytes in aqueous solutions, especially for symmetric systems (systems in which both polyions have similar molecular weight and concentration). However, in recent years, the complexation of polyelectrolytes with alternative building blocks such as small charged molecules (multivalent inorganic species, oligopeptides, and oligoamines, among others) has gained attention in different areas. In this review, we discuss the physicochemical characteristics of the complexes formed by polyelectrolytes and multivalent small molecules, putting a special emphasis on their similarities with the well-known polycation-polyanion complexes. In addition, we analyze the potential of these complexes to act as versatile functional platforms in various technological fields, such as biomedicine and advanced materials engineering.

Ultrasound-guided lateral pericapsular hip desensitization of the articular branches of the cranial gluteal nerve: A canine cadaveric study and feasibility study in dogs.

OBJECTIVE: To develop and assess the feasibility, as a diagnostic block, of an ultrasound-guided lateral pericapsular hip desensitization (L-PHD) technique in dogs. STUDY DESIGN: Prospective, randomized, anatomical and feasibility study. ANIMALS: A total of 11 canine cadavers and eight adult dogs scheduled for acetabular surgical denervation. METHODS: After studying the ultrasound anatomy of the lateral aspect of the gluteal region and determining an acoustic window to perform an ultrasound-guided L-PHD in three canine cadavers, the right and left hemipelves of eight canine cadavers were injected in the interfascial plane located lateral (LL-PHD group) or medial (LM-PHD group) to the deep gluteal muscle, with 0.05 mL kg-1 of dye per hip on each cadaver. The staining of the pericapsular nerves was assessed by anatomical dissection. Then, the LM-PHD was performed using 2% lidocaine as a diagnostic block in dogs scheduled for acetabular surgical denervation. Positive predictive value (PPV) was calculated for those animals who had favorable outcomes after acetabular surgical denervation. RESULTS: The ultrasound-guided LL-PHD and LM-PHD could be performed by inserting the needle lateral and medial to the deep gluteal muscle. Ultrasound-guided LL-PHD stained the cranial gluteal nerve and its muscular branches in all injections and partially stained the lumbosacral trunk in two out of eight cadavers. The LM-PHD selectively stained the articular branches of the cranial gluteal nerve in all but one cadaver. The PPV for LM-PHD successful test prediction was 85.7% (95% confidence interval: 48.6% to 98.6%). CONCLUSIONS: and clinical significance Ultrasound-guided LM-PHD using 0.05 mL kg-1 of dye selectively stained the articular branches of the cranial gluteal nerve in canine cadavers. The LM-PHD technique is feasible and could be used as a diagnostic block before acetabular surgical denervation in dogs.

Lumbosacral plexus block using a combination of ultrasound-guided lateral pre-iliac and parasacral approaches in cats.

OBJECTIVE: To describe an ultrasound-guided lateral pre-iliac (LPI) and parasacral (PS) approach in feline cadavers (phase I) and compare the perioperative analgesic use and complications in cats administered LPI and PS blocks (group PNB) or epidural anesthesia (group EPI) for pelvic limb surgery (phase II). STUDY DESIGN: Experimental uncontrolled, anatomic and retrospective cohort study. ANIMALS: A group of eight feline cadavers and 52 medical records. METHODS: Bilateral LPI and PS approaches with 0.1 mL kg-1 of dye to stain the femoral and obturator nerves and the lumbosacral trunk, respectively, were performed on each cadaver. Nerve staining effect was evaluated upon dissections (phase I). Perioperative analgesics use, and complication rates were retrospectively compared between groups PNB and EPI (phase II). Continuous data were compared using the Mann-Whitney U test and the prevalence of events with Fisher's exact test. Differences were considered significant when p < 0.05. RESULTS: Dissections revealed that the LPI approach stained 94% and 75% of the femoral and obturator nerves, respectively. The PS approach stained 100% of the lumbosacral trunks. Cats enrolled in group PNB (n = 23) were administered lower doses of intraoperative opioids than those in group EPI (n = 25) (p = 0.006). Intraoperative rescue analgesia was required in 60% and 17.4% of cats enrolled in groups EPI and PNB, respectively (p = 0.003). Group PNB required more intraoperative anticholinergics than group EPI (p = 0.02). There were no differences in postoperative pain scores, analgesic use and complication rates. CONCLUSIONS AND CLINICAL RELEVANCE: The ultrasound-guided LPI and PS approach stained the femoral/obturator nerves and the lumbosacral trunk, respectively, in feline cadavers. Furthermore, PNB was associated with lower intraoperative opioid use and similar postoperative pain and analgesic use compared with epidural anesthesia in a cohort of cats undergoing surgery of the pelvic limb.

Pericapsular hip desensitization in dogs: a cadaveric study and case series.

OBJECTIVE: To develop and assess the efficacy of an ultrasound (US)-guided pericapsular hip desensitization (PHD) technique in dogs. STUDY DESIGN: Prospective, randomized, anatomical study and a case series. ANIMALS: A total of 30 healthy dogs, eight canine cadavers and seven dogs with hip osteoarthritis. METHODS: After studying the US anatomy of the medial aspect of the coxofemoral joint and determining an acoustic window to perform an US-guided PHD in healthy dogs, the US-guided PHD was performed bilaterally in canine cadavers. A low [(LV) 0.1 mL kg-1] and high [(HV) 0.2 mL kg-1] volume of dye was injected per hip on each cadaver. The staining of the pericapsular nerves was assessed by anatomical dissection, and comparison between LV and HV was assessed using Fisher's exact test. Then, the US-guided PHD was performed using a triamcinolone-bupivacaine solution in dogs with hip osteoarthritis. Dynamic pain response was assessed before and after injection. The canine brief pain inventory (CBPI) questionnaire was used to assess treatment efficacy and duration. RESULTS: The US-guided PHD could be performed by inserting the needle between the iliopsoas muscle and the periosteum of the ilium. The articular branches of the femoral and obturator nerves were stained in all cadavers using both volumes. The main femoral nerve was never stained, but the main obturator nerve was stained in 37.5% and 100% of injections using LV and HV, respectively (p = 0.026). Treated animals showed decreased dynamic pain response after the injection. Compared with baseline, CBPI scores were reduced by ≥ 50% for ≥ 12 weeks in all but one dog. CONCLUSIONS AND CLINICAL SIGNIFICANCE: The US-guided PHD with both 0.1 and 0.2 mL kg-1 volumes stained the articular branches of the femoral and obturator nerves in canine cadavers and was associated with clinical improvement in dogs with hip osteoarthritis.

CT for Evaluation of Hemoptysis.

Hemoptysis, which is defined as expectoration of blood from the alveoli or airways of the lower respiratory tract, is an alarming clinical symptom with an extensive differential diagnosis. CT has emerged as an important noninvasive tool in the evaluation of patients with hemoptysis, and the authors present a systematic but flexible approach to CT interpretation. The first step in this approach involves identifying findings of parenchymal and airway hemorrhage. The second step is aimed at determining the mechanism of hemoptysis and whether a specific vascular supply can be implicated. Hemoptysis can have primary vascular and secondary vascular causes. Primary vascular mechanisms include chronic systemic vascular hypertrophy, focally damaged vessels, a dysplastic lung parenchyma with systemic arterial supply, arteriovenous malformations and fistulas, and bleeding at the capillary level. Evaluating vascular mechanisms of hemoptysis at CT also entails determining if a specific vascular source can be implicated. Although the bronchial arteries are responsible for most cases of hemoptysis, nonbronchial systemic arteries and the pulmonary arteries are important potential sources of hemoptysis that must be recognized. Secondary vascular mechanisms of hemoptysis include processes that directly destroy the lung parenchyma and processes that directly invade the airway. Understanding and employing this approach allow the diagnostic radiologist to interpret CT examinations accurately in patients with hemoptysis and provide information that is best suited to directing subsequent treatment. ©RSNA, 2021.

Ultrasound-guided dorsal approach for the brachial plexus block in common kestrels (Falco tinnunculus): a cadaver study.

OBJECTIVE: To develop an ultrasound-guided dorsal approach to the brachial plexus and to investigate the nerve distribution and staining of a dyed injectate in common kestrel (Falco tinnunculus) cadavers. STUDY DESIGN: Prospective, cadaver study. ANIMALS: A group of three common kestrel cadavers (six wings). METHODS: All cadavers were fresh-frozen at -20 °C and thawed for 10 hours at room temperature before the study. The cadavers were placed in sternal recumbency and their wings were abducted. A 8-13 MHz linear-array transducer was placed over the scapulohumeral joint, at the centre of a triangle formed by the scapula and the humerus. The brachial plexus was identified between the scapulohumeralis muscle and the pectoralis major muscle, as hypoechoic structures lying just cranially to the axillary vessels. After ultrasound-guided brachial plexus identification, a 22 gauge, 50 mm insulated needle was advanced in-plane using ultrasound visualization. A volume of 0.5 mL kg-1 of a 3:1 (2% lidocaine:methylene blue) solution was injected. Following cadaver dissection, the pattern of the spread was assessed, and the extent of nerve staining was measured with a calliper and deemed adequate if more than 0.6 cm of the nerve staining was achieved. RESULTS: The brachial plexus was clearly identified in all wings with the dorsal approach. After dye injection, all the branches of the brachial plexus defined as nerves 1-5 (N1, N2, N3, N4 and N5) were completely stained in five (83%) and partially stained in one (17%) of the six wings. CONCLUSIONS AND CLINICAL RELEVANCE: The ultrasound-guided dorsal approach allows a clear visualization of the brachial plexus structure. The injection of 0.5 mL kg-1of a lidocaine/dye solution produced complete nerve staining in most cases. Further in vivo studies are mandatory to confirm the clinical efficacy of this locoregional anaesthesia technique in common kestrels (Falco tinnunculus).

Transversus abdominis plane block in cat cadavers: anatomical description and comparison of injectate spread using two- and three-point approaches.

OBJECTIVE: To describe the sonoanatomy of the abdominal wall in live cats and to compare the distribution pattern of two versus three ultrasound-guided transversus abdominis plane (TAP) injections using clinically applicable volumes of lidocaine-dye solution in cat cadavers. STUDY DESIGN: Prospective anatomical study. ANIMALS: A total of eight client-owned healthy cats and eight cat cadavers. METHODS: Ultrasound anatomy of the abdominal wall, landmarks and sites for needle access were determined in live cats. Ultrasound-guided TAP injections were performed in eight thawed cat cadavers. Volumes of 0.25 or 0.16 mL kg-1 per point of a lidocaine-dye solution were injected using either two [subcostal and preiliac (SP)] or three [subcostal, retrocostal and preiliac (SRP)] injection points, respectively. Each cadaver was then dissected to determine the injectate distribution and the number of thoracolumbar nerves stained with each approach. The target nerves were defined as the ventromedial branches of the thoracic nerves 10 (T10), T11, T12, T13 and lumbar nerves 1 (L1) and L2. RESULTS: Sonoanatomy was consistent with anatomy upon dissection and the TAP was identified in all cadavers. A total of 16 subcostal, 16 preiliac and nine retrocostal TAP injections were performed. The overall staining success rate of the target nerves was 66.7% and 92.6% for the SP and SPR approaches, respectively (p = 0.02). The ventromedial branches of T10, T11, T12, T13, L1 and L2 were stained in 57.1%, 100.0%, 85.7%, 28.6%, 42.9% and 85.7%, and in 66.7%, 100.0%, 100.0%, 100.0%, 88.9% and 100.0% of the cases with the SP and SRP approaches, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: The SRP approach allowed a broader distribution around the target nerves, whereas a staining gap was observed at T13 and L1 with the SP approach. Further studies are necessary to investigate the analgesic effect of these approaches in a clinical setting.

Insulin Delivery from Glucose-Responsive, Self-Assembled, Polyamine Nanoparticles: Smart "Sense-and-Treat" Nanocarriers Made Easy.

Polyamine-salt aggregates (PSA) are biomimetic soft materials that have attracted great attention due to their straightforward fabrication methods, high drug-loading efficiencies, and attractive properties for pH-triggered release. Herein, a simple and fast multicomponent self-assembly process was used to construct cross-linked poly(allylamine hydrochloride)/phosphate PSAs (hydrodynamic diameter of 360 nm) containing glucose oxidase enzyme, as a glucose-responsive element, and human recombinant insulin, as a therapeutic agent for the treatment of diabetes mellitus (GI-PSA). The addition of increasing glucose concentrations promotes the release of insulin due to the disassembly of the GI-PSAs triggered by the catalytic in situ formation of gluconic acid. Under normoglycemia, the GI-PSA integrity remained intact for at least 24 h, whereas hyperglycemic conditions resulted in 100 % cargo release after 4 h of glucose addition. This entirely supramolecular strategy presents great potential for the construction of smart glucose-responsive delivery nanocarriers.

Cigarette smoke-induced changes in the murine vocal folds: a Raman spectroscopic observation.

Raman spectroscopic methods are being projected as novel tools to study the early invisible molecular level changes in a label-free manner. In the present study, we have used Raman spectroscopy to explore the earliest biochemical changes in murine vocal folds in response to time-bound cigarette smoke exposure. Mice were exposed to cigarette smoke for 2 or 4-weeks through a customized smoke inhalation system. The larynx was collected and initial evaluations using standard methods of analysis such as histopathology and immunofluorescence was performed. Concurrent unstained sections were used for Raman imaging. Two common pathological features of vocal fold disorders including alterations in collagen content and epithelial hypercellularity, or hyperplasia, were observed. The mean spectra, principal component analysis, and Raman mapping also revealed differences in the collagen content and hypercellularity in the smoke exposed tissues. The differences in 2-week exposed tissues were found to be more prominent as compared to 4-week. This was attributed to adaptive responses and the already reported biphasic effects, which suggest that collagen synthesis is significantly reduced at higher cigarette smoke concentrations. Overall findings of the study are supportive of the prospective application of Raman imaging in monitoring changes due to cigarette smoke in the vocal folds.

Cobalt Corroles as Electrocatalysts for Water Oxidation: Strong Effect of Substituents on Catalytic Activity.

Two Co(III) complexes (1Py2 and 2Py2) of new corrole ligands H3L1 (5,15-bis(p-methylcarboxyphenyl)-10-(o-methylcarboxyphenyl)corrole) and H3L2 (5,15-bis(p-nitrophenyl)-10-(o-methylcarboxyphenyl)corrole) with two apical pyridine ligands have been synthesized and thoroughly characterized by cyclic voltammetry, UV-vis-NIR, and EPR spectroscopy, spectroelectrochemistry, single-crystal X-ray diffraction studies, and DFT methods. Complexes 1Py2 and 2Py2 possess much lower oxidation potentials than cobalt(III)-tris-pentafluorophenylcorrole (Co(tpfc)) and similar corroles containing pentafluorophenyl (C6F5) substituents, thus allowing access to high oxidation states of the former metallocorroles using mild chemical oxidants. The spectroscopic (UV-vis-NIR and EPR) and electronic properties of several oxidation states of these complexes have been determined by a combination of the mentioned methods. Complexes 1Py2 and 2Py2 undergo three oxidations within 1.3 V vs FcH+/FcH in MeCN, and we show that both complexes catalyze water oxidation in an MeCN/H2O mixture upon the third oxidation, with kobs (TOF) values of 1.86 s-1 at 1.29 V (1Py2) and 1.67 s-1 at 1.37 V (2Py2). These values are five times higher than previously reported TOF values for C6F5-substituted cobalt(III) corroles, a finding we ascribe to the additional charge in the corrole macrocycle due to the increased oxidation state. This work opens up new possibilities in the study of metallocorrole water oxidation catalysts, particularly by allowing spectroscopic probing of high-oxidation states and showing strong substituent-effects on catalytic activity of the corrole complexes.

Redox-active polyamine-salt aggregates as multistimuli-responsive soft nanoparticles.

Polyamine-salt aggregates have become promising soft materials in nanotechnology due to their easy preparation process and pH-responsiveness. Here, we report the use of hexacyanoferrate(ii) and hexacyanoferrate(iii) as electroactive crosslinking agents for the formation of nanometer-sized redox-active polyamine-redox-salt aggregates (rPSA) in bulk suspension. This nanoplatform can be selectively assembled or disassembled under different stimuli such as redox environment, pH and ionic strength. By changing the charge of the building blocks, external triggers allow switching the system between two phase states: aggregate-free solution or colloidal rPSA dispersion. The stimuli-activated modulation of the assembly/disassembly processes opens a path to exploit rPSA in technologies based on smart nanomaterials.

Development of a lateral ultrasound-guided approach for the proximal radial, ulnar, median and musculocutaneous (RUMM) nerve block in cats.

OBJECTIVE: To describe a lateral ultrasound (US)-guided approach to the radial, ulnar, median and musculocutaneous (RUMM) nerves through a single proximal in-plane insertion in cats and to determine whether one or two injection points are required to successfully stain all the target nerves. STUDY DESIGN: Prospective study. ANIMALS: A total of eight client-owned healthy cats and 12 cat cadavers. METHODS: In live cats, the US anatomy of the brachium, the landmarks and the site for needle accesses were determined. Then, 12 thawed feline cadavers were used to assess the spread of dye solution and nerve staining following the US-guided proximal-lateral-humeral RUMM injection using one and two injection points. Each cadaver was injected with 0.15 mL kg-1 of a 0.25% new methylene blue solution in either a single injection aimed for the radial nerve of one limb (G1) or via two sites delivering 0.1 mL kg-1 and 0.05 mL kg-1 aimed for the radial and musculocutaneous nerves of the opposite limb, respectively (G2). Upon dissection, staining of the target nerves around their circumference for length of >1 cm was considered successful. RESULTS: Sonoanatomy was consistent with anatomy upon dissection and target nerves were identified in all cadavers. Staining was 100% successful for the radial, median and ulnar nerves in both groups, and 41.7% and 100% for the musculocutaneous nerve in G1 and G2, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: This novel lateral US-guided approach for the proximal RUMM nerve block allowed a good identification of the nerves and related structures, and it provided a consistent muscular structure through which the needle could be easily guided. An injection performed in two aliquots (within the caudal and cranial compartments of the neurovascular sheath) appeared to be necessary to successfully stain all the target nerves.

Polyamine Colloids Cross-Linked with Phosphate Ions: Towards Understanding the Solution Phase Behavior.

Ionically crosslinked poly(allylamine)/phosphate (PAH/Pi) colloids consist of self-assembled nanostructures stabilized by supramolecular interactions. Under physiological conditions, these interactions should be present at high ionic strength and only in a narrow pH window to be effective as drug delivery agents. In this work we study the effect of the pH and ionic strength in the chemical behaviour of inorganic phosphate (Pi), poly(allylamine hydrochloride) (PAH) and their mixture in aqueous solution (PAH-Pi). By combination of experimental measurements and a theoretical model, we demonstrate that the driving force that leads to the formation of colloids is the electrostatic pairing between the positively charged amino groups in PAH and negatively charged HPO42- ions. Increasing the ionic strength of the system by addition of KCl weakens the PAH-Pi interactions and narrows the pH stability window from 4 to 1.8 pH units. In addition, a fully reversible system was obtained in which the colloids assemble and disassemble by changing the pH between 6.8 and 7.1 at high ionic strength, making them suitable for use as pH-responsive nanocarriers.

Metal Nanoparticle Enhancement of Electron Transfer to Tethered Redox Centers through Self-Assembled Molecular Films.

Metal-nanoparticle-mediated electron transfer (ET) across an insulator thin film containing nanoparticles with attached redox centers was studied using electrochemical impedance spectroscopy. Specifically, a gold spherical microelectrode was modified with 16-amino-1-hexa-decanethiol, creating an insulator film. This was followed by the electrostatic adsorption of gold nanoparticles and the covalent attachment of Os2+ redox centers. A variation of the Creager-Wooster method was developed to get quantitative information regarding the ET kinetics of the system. The experimental data obtained from a single measurement was fitted with a model that decouples two or more ET processes with different time constants and considers a Gaussian distribution of tunneling distances. Two parallel ET mechanisms were observed: one in which the electrons flow by tunneling between the surface and the redox couples with a low kET0 = 1.3 s-1 and a second one in which an enhancement of the electron transfer is produced due to the presence of the gold nanoparticles with a kET0 = 7 × 104 s-1. In this study, we demonstrate that the gold nanoparticle electron transfer enhancement is present only in the local environment of the nanoparticle, showing that the nanoscale architecture is crucial to maximize the enhancement effect.

Continuous assembly of supramolecular polyamine-phosphate networks on surfaces: preparation and permeability properties of nanofilms.

Supramolecular self-assembly of molecular building blocks represents a powerful "nanoarchitectonic" tool to create new functional materials with molecular-level feature control. Here, we propose a simple method to create tunable phosphate/polyamine-based films on surfaces by successive assembly of poly(allylamine hydrochloride) (PAH)/phosphate anions (Pi) supramolecular networks. The growth of the films showed a great linearity and regularity with the number of steps. The coating thickness can be easily modulated by the bulk concentration of PAH and the deposition cycles. The PAH/Pi networks showed chemical stability between pH 4 and 10. The transport properties of the surface assemblies formed from different deposition cycles were evaluated electrochemically by using different redox probes in aqueous solution. The results revealed that either highly permeable films or efficient anion transport selectivity can be created by simply varying the concentration of PAH. This experimental evidence indicates that this new strategy of supramolecular self-assembly can be useful for the rational construction of single polyelectrolyte nanoarchitectures with multiple functionalities.

Nanoscopic Characterisation of Individual Endogenous Protein Aggregates in Human Neuronal Cells.

The aberrant misfolding and subsequent conversion of monomeric protein into amyloid aggregates characterises many neurodegenerative disorders, including Parkinson's and Alzheimer's diseases. These aggregates are highly heterogeneous in structure, generally of low abundance and typically smaller than the diffraction limit of light (≈250 nm). To overcome the challenges these characteristics pose to the study of endogenous aggregates formed in cells, we have developed a method to characterise them at the nanometre scale without the need for a conjugated fluorophore. Using a combination of DNA PAINT and an amyloid-specific aptamer, we demonstrate that this technique is able to detect and super-resolve a range of aggregated species, including those formed by α-synuclein and amyloid-ß. Additionally, this method enables endogenous protein aggregates within cells to be characterised. We found that neuronal cells derived from patients with Parkinson's disease contain a larger number of protein aggregates than those from healthy controls.