Preferential Myosin Heavy Chain Isoform B Expression May Contribute to the Faster Velocity of Contraction in Veins versus Arteries

Smooth muscle myosin heavy chains occur in 2 isoforms, SMA (slow) and SMB (fast). We hypothesized that the SMB isoform is predominant in the faster-contracting rat vena cava compared to thoracic aorta. We compared the time to half maximal contraction in response to a maximal concentration of endothelin-1 (ET-1; 100 nM), potassium chloride (KCl; 100 mM) and norepinephrine (NE; 10 µM). The time to half maximal contraction was shorter in the vena cava compared to aorta (aorta: ET-1 = 235.8 ± 13.8 s, KCl = 140.0 ± 33.3 s, NE = 19.8 ± 2.7 s; vena cava: ET-1 = 121.8 ± 15.6 s, KCl = 49.5 ± 6.7 s, NE = 9.0 ± 3.3 s). Reverse-transcription polymerase chain reaction supported the greater expression of SMB in the vena cava compared to aorta. SMB was expressed to a greater extent than SMA in the vessel wall of the vena cava. Western analysis determined that expression of SMB, relative to total smooth muscle myosin heavy chains, was 12.5 ± 4.9-fold higher in the vena cava compared to aorta, while SMA was 4.9 ± 1.2-fold higher in the aorta than vena cava. Thus, the SMB isoform is the predominant form expressed in rat veins, providing one possible mechanism for the faster response of veins to vasoconstrictors

Decoration of nanovesicles with pH (low) insertion peptide (pHLIP) for targeted delivery

Acidity at surface of cancer cells is a hallmark of tumor microenvironments, which does not depend on tumor perfusion, thus it may serve as a general biomarker for targeting tumor cells. We used the pH (low) insertion peptide (pHLIP) for decoration of liposomes and niosomes. pHLIP senses pH at the surface of cancer cells and inserts into the membrane of targeted cells, and brings nanomaterial to close proximity of cellular membrane. DMPC liposomes and Tween 20 or Span 20 niosomes with and without pHLIP in their coating were fully characterized in order to obtain fundamental understanding on nanocarrier features and facilitate the rational design of acidity sensitive nanovectors. The samples stability over time and in presence of serum was demonstrated. The size, ζ-potential, and morphology of nanovectors, as well as their ability to entrap a hydrophilic probe and modulate its release were investigated. pHLIP decorated vesicles could be useful to obtain a prolonged (modified) release of biological active substances for targeting tumors and other acidic diseased tissues

The nasal delivery of nanoencapsulated statins – An approach for brain delivery

© 2016 Clementino et al. Purpose: Along with their cholesterol-lowering effect, statins have shown a wide range of pleiotropic effects potentially beneficial to neurodegenerative diseases. However, such effects are extremely elusive via the conventional oral administration. The purpose of the present study was to prepare and characterize the physicochemical properties and the in vivo biodistribution of simvastatin-loaded lecithin/chitosan nanoparticles (SVT-LCNs) suitable for nasal administration in view of an improved delivery of the statins to the brain. Materials and methods: Chitosan, lecithin, and different oil excipients were used to prepare nanocapsules loaded with simvastatin. Particle size distribution, surface charge, structure, simvastatin loading and release, and interaction with mucus of nanoparticles were determined. The nanoparticle nasal toxicity was evaluated in vitro using RPMI 2651 nasal cell lines. Finally, in vivo biodistribution was assessed by gamma scintigraphy via Tc99m labeling of the particles. Results: Among the different types of nanoparticles produced, the SVT-LCN_MaiLab showed the most ideal physicochemical characteristics, with small diameter (200 nm), positive surface charge (+48 mV) and high encapsulation efficiency (EE; 98%). Size distribution was further confirmed by nanoparticle tracking analysis and electron microscopy. The particles showed a relatively fast release of simvastatin in vitro (35.6%±4.2% in 6 hours) in simulated nasal fluid. Blank nanoparticles did not show cytotoxicity, evidencing that the formulation is safe for nasal administration, while cytotoxicity of simvastatin-loaded nanoparticles (IC50) was found to be three times lower than the drug solution (9.92 vs 3.50 μM). In rats, a significantly higher radioactivity was evidenced in the brain after nasal delivery of simvastatin-loaded nanoparticles in comparison to the administration of a similar dose of simvastatin suspension. Conclusion: The SVT-LCNs developed presented some of the most desirable characteristics for mucosal delivery, that is, small particle size, positive surface charge, long-term stability, high EE, and mucoadhesion. In addition, they displayed two exciting features: First was their biodegradability by enzymes present in the mucus layer, such as lysozyme. This indicates a new Trojan-horse strategy which may enhance drug release in the proximity of the nasal mucosa. Second was their ability to enhance the nose-to-brain transport as evidenced by preliminary gamma scintigraphy studies

NUP98-fusion transcripts characterize different biological entities within acute myeloid leukemia: A report from the AIEOP-AML group.

In the last years, collaborative studies have joined to link the degree of genetic heterogeneity of acute myeloid leukemia (AML) to clinical outcome,1, 2 allowing risk stratification before therapy and guiding post-induction treatment of children with AML. So far, still half of these patients, whose disease is usually characterized by a grim prognosis, lack a known biomarker offering opportunities of targeted treatment

Energy absorption in actual tractor rollovers with different tire configurations

In order to better understand the complexities of modern tractor rollover, this paper investigates the energy absorbed by a Roll-Over Protective Structure (ROPS) cab during controlled lateral rollover testing carried out on a modern narrow-track tractor with a silent-block suspended ROPS cab. To investigate how different tractor set-ups may influence ROPS and energy partitioning, tests were conducted with two different wheel configurations, wide (equivalent to normal ‘open field’ operation) and narrow (equivalent to ‘orchard/vineyard’ operation), and refer to both the width of the tires and the corresponding track. Dynamic load cells and displacement transducers located at the ROPS-ground impact points provided a direct measurement of the energy absorbed by the ROPS cab frame. A trilateration method was developed and mounted onboard to measure load cell trajectory with respect to the cab floor in real-time. The associated video record of each rollover event provided further information and opportunity to explain the acquired data. The narrow tire configuration consistently subjected the ROPS cab frame to more energy than the wide tire arrangement. To better evaluate the influence of the ROPS cab silent-blocks in lateral rollover, static and dynamic tests were performed. The results confirm that tires influence the energy partition significantly and that further understanding of silent-blocks’ dynamic performance is warranted

Eculizumab treatment: stochastic occurrence of C3 binding to individual PNH erythrocytes

C5 blockade by eculizumab prevents complement-mediated intravascular hemolysis in paroxysmal nocturnal hemoglobinuria (PNH). However, C3-bound PNH red blood cells (RBCs), arising in almost all treated patients, may undergo extravascular hemolysis reducing clinical benefits. Despite the uniform deficiency of CD55 and of CD59, there are always two distinct populations of PNH RBCs, with (C3+) and without (C3-) C3 binding

Model cell membrane interaction with a bioinspired amphoteric polymer

We present recent investigation by means of nanoscale techniques on biocompatible linear polyamidoamines with amphoteric character, namely AGMA1 and ARGO7. These polymers have been shown of extremely promising and already proved medical interest, comprising their strong protection actions against virus infection, mainly papilloma and herpes and the extremely low toxicity of their DNA complexes, with respect to other used polymers such as PEI and protamine, applied in nanovector design for gene delivery. Our studies focus on the most important of these polymers, AGMA1, a prevailingly cationic 4-aminobutylguanidine-deriving PAA, whose mechanism of action is so far not fully understood. The current understanding is that its interaction with cell surfaces by means of glycosaminoglycans (HSPG) has a major role in its protective action against viruses. Yet, AGMA1 is active also against HPV-31, whose attachment does not appear to be dependent on HSPG. HPV-31, whose attachment does not appear to be dependent on HSPG. Therefore, AGMA1 binds other (as yet unidentified) receptors on the cell surface. As the known recipient is the HS carbohydrate moiety, other sugars rich membrane components have been proposed as probable AGMA1 target. Therefore, to shed a light on the mechanism of interaction of the polymer with sugar containing biologically relevant molecules, not HS, we have investigated AGMA1 in interaction with glycophyngolipids, Specifically, we studied multicomponent symmetric vesicles enriched in ganglioside GM1 built to mimic biological membrane domains, in the presence of AGMA1, At physiological pH, electrostatic effects should be the relevant interactions between GM1 and AGMA1. Taking advantage of the same mechanism we investigated the possibility of building lipid based core-shell particles to vehiculate AGMA1/siRNA complexes. Moreover, since it is probable that AGMA1 interacts with the barrier of mucus which cover the involved tissue we have extended our investigations also to mucin, constituting the biological barrier to the target tissues of the medical application of the polymers

Model cell membrane interaction with a bioinspired amphoteric polymer

We present recent investigation by means of nanoscale techniques on biocompatible linear polyamidoamines with amphoteric character, namely AGMA1 and ARGO7. These polymers have been shown of extremely promising and already proved medical interest, comprising their strong protection actions against virus infection, mainly papilloma and herpes and the extremely low toxicity of their DNA complexes, with respect to other used polymers such as PEI and protamine, applied in nanovector design for gene delivery. Our studies focus on the most important of these polymers, AGMA1, a prevailingly cationic 4-aminobutylguanidine-deriving PAA, whose mechanism of action is so far not fully understood. The current understanding is that its interaction with cell surfaces by means of glycosaminoglycans (HSPG) has a major role in its protective action against viruses. Yet, AGMA1 is active also against HPV-31, whose attachment does not appear to be dependent on HSPG. HPV-31, whose attachment does not appear to be dependent on HSPG. Therefore, AGMA1 binds other (as yet unidentified) receptors on the cell surface. As the known recipient is the HS carbohydrate moiety, other sugars rich membrane components have been proposed as probable AGMA1 target. Therefore, to shed a light on the mechanism of interaction of the polymer with sugar containing biologically relevant molecules, not HS, we have investigated AGMA1 in interaction with glycophyngolipids, Specifically, we studied multicomponent symmetric vesicles enriched in ganglioside GM1 built to mimic biological membrane domains, in the presence of AGMA1, At physiological pH, electrostatic effects should be the relevant interactions between GM1 and AGMA1. Taking advantage of the same mechanism we investigated the possibility of building lipid based core-shell particles to vehiculate AGMA1/siRNA complexes. Moreover, since it is probable that AGMA1 interacts with the barrier of mucus which cover the involved tissue we have extended our investigations also to mucin, constituting the biological barrier to the target tissues of the medical application of the polymers

Interaction of mucins with bioinspired polymers and drug delivery particles

Mucins are glycoproteins with high molecular weight and an abundance of negatively charged oligosaccharide side chains, representing the main components in the mucous gels apart from water. Mucin structure consists of a flexible backbone (mainly serine and threonine residues) which serves as anchoring points for oligosaccharide side chains, and hydrophobic \u201cnaked domains\u201d enriched in cysteine residues. The latter can form inter-molecular bonds via disulphide links, promoting mucin association in solution. Therefore, mucins can establish adhesive interactions with particulates/biomacromolecules via electrostatic interactions, van der Waals forces, hydrophobic forces, hydrogen bonding, or chain entanglement. Mucosal drug delivery vehicles can either penetrate rapidly or establish prolonged contact. However, their development is of great challenge because little is still known about the interactions between mucin and other macromolecules. We are currently working on a comprehensive study of the interaction between mucin and macromolecules of interest for pharmaceutical developments by complementary techniques. To this scope, we employ biocompatible natural and synthetic polymers with different physical-chemical characteristics. Among them, linear polyamidoamines with amphoteric character are particularly interesting for their cyto-biocompatibility. It is indeed crucial to characterise such interactions not only in the bulk but also at the interface, since complexation between mucins and biomacromolecules takes place close to the cell membrane surface. Moreover, the strategy to overcome mucus barrier and achieve long retention time in the cell surface is to develop nano-agents which can effectively penetrate the mucus layer and accumulate at the epithelial surface. In this framework we present preliminary investigations in the bulk by small angle x-ray scattering (SAXS) and at the solid-liquid interface by employing quartz crystal microbalance (QCM-D)

Building a biomimetic membrane for neutron reflectivity investigation : complexity, asymmetry and contrast

The preparation and investigation of model membranes is deserving growing interest both for the physics of complex systems, and for biology. The need of simplified models should preserve mimicking the qualifying characteristics of biological membranes, and keep non-invasive and detailed description. As a main feature, biological membranes are non-homogeneous in the disposition of components, both in the lateral and in the transverse direction. We prepared asymmetric supported membranes containing GM1 ganglioside in biomimetic proportion according to different protocols. Then, we studied their internal structure by neutron reflectometry, providing few-Angstrom sensitivity in the cross direction meanwhile avoiding radiation damage. This technique can also be profitably applied to study interactions at the membrane surface. The best protocol has proven to be the Langmuir-Blodgett/Langmuir-Schaefer depositions. Notably, also the simpler and most accessible protocol of vesicle fusion was found to be suitable for straightforward and good quality deposition of compositionally asymmetric membranes