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Biomedical subjects

Israel Rubinstein

Publications and source records attributed to Israel Rubinstein.

At least 19 recordsLinked to original sources

Hog barn dust extract increases macromolecular efflux from the hamster cheek pouch.

The purpose of this study was to determine whether short-term exposure to an aqueous extract of hog barn dust increases macromolecular efflux from the intact hamster cheek pouch and, if so, to begin to determine the mechanism(s) underlying this response. By using intravital microscopy, we found that suffusion of hog barn dust extract onto the intact hamster cheek pouch for 60 min elicited a significant, concentration-dependent leaky site formation and increase in clearance of FITC-labeled dextran (molecular mass, 70 kDa). This response was significantly attenuated by suffusion of catalase (60 U/ml), but not by heat-inactivated catalase, and by pretreatment with dexamethasone (10 mg/kg iv) (P < 0.05). Catalase had no significant effects on adenosine-induced increase in macromolecular efflux from the cheek pouch. Suffusion of hog barn dust extract had no significant effects on arteriolar diameter in the cheek pouch. Taken together, these data indicate that hog barn dust extract increases macromolecular efflux from the in situ hamster cheek pouch, in part, through local elaboration of reactive oxygen species that are inactivated by catalase. This response is specific and attenuated by corticosteroids. We suggest that plasma exudation plays an important role in the genesis of upper airway dysfunction evoked by short-term exposure to hog barn dust.

Adenosine↗

Intratracheal and subcutaneous liposomal VIP normalizes arterial pressure in spontaneously hypertensive hamsters.

We determined whether a single intratracheal and subcutaneous administration of biocompatible and biodegradable vasoactive intestinal peptide self-associated with sterically stabilized liposomes (VIP-SSL) normalizes mean arterial pressure (MAP) in spontaneously hypertensive hamsters (SHH). We found that VIP-SSL (0.1 nmol) administered by either routes normalizes MAP (p < 0.05). Maximal effect was observed within 10-20 min and lasted for 6 h. VIP-SSL had no significant effects on heart rate. VIP alone (0.1 nmol) and empty SSL had no significant effects on MAP. VIP-SSL (0.1 nmol) had no significant effects on MAP and heart rate in age/genetically-matched control hamsters. Given these data, we suggest that pulmonary and subcutaneous delivery of VIP-SSL should be further developed as peptide nanomedicine for essential hypertension.

Animals↗

Assembly of coordination nanostructures via ligand derivatization of oxide surfaces.

A scheme is presented for the construction of coordination nanostructures on oxide surfaces (glass, Si/SiO2, quartz), based on application of epoxy-terminated monolayers as anchors for covalent grafting of ligands. Two ligands bearing amine groups were reacted with epoxysilane monolayers on oxide surfaces, providing ligand-terminated substrates. The ligands employed were (i) a pyridine moiety, used for subsequent binding of cobalt tetraphenylporphine (CoTPP), and (ii) deferoxamine (DFX), which contains hydroxamic acid moieties, used for subsequent construction of various Zr4+-based coordination layers. The results suggest that a dense ligand layer was obtained in both cases, allowing the formation of coordination overlayers on the oxide surfaces. The growth of coordinated layers was similar to analogous overlayers assembled on Au substrates, indicating that high ligand coverage is achieved by the epoxy-amine surface reaction. Epoxy-based functionalization of oxide substrates is a mild and efficient method for preparing high-quality coordination overlayers. Moreover, the method makes use of commercially available silane and amine reactants, providing the basis for wide application.

Journal Article↗

Branched coordination multilayers on gold.

A C3-symmetric tridentate hexahydroxamate ligand molecule was specially synthesized and used for coordination self-assembly of branched multilayers on Au surfaces precoated with a self-assembled monolayer (SAM) of ligand anchors. Layer-by-layer (LbL) growth of multilayers via metal-organic coordination using Zr4+ ions proceeds with high regularity, adding one molecular layer in each step, as shown by ellipsometry, wettability, UV-vis spectroscopy, and atomic force microscopy (AFM). The branched multilayer films display improved stiffness, as well as a unique defect self-repair capability, attributed to cross-linking in the layers and lateral expansion over defects during multilayer growth. Transmetalation, i.e., exposure of Zr4+-based assemblies to Hf4+ ions, was used to evaluate the cross-linking. Conductive atomic force microscopy (AFM) was used to probe the electrical properties of the multilayers, revealing excellent dielectric behavior. The special properties of the branched layers were emphasized by comparison with analogous multilayers prepared similarly using linear (tetrahydroxamate) ligand molecules. The process of defect annihilation by bridging over defective areas, attributed to lateral expansion via the excess bishydroxamate groups, was demonstrated by introduction of artificial defects in the anchor monolayer, followed by assembly of two layers of either the linear or the branched molecule. Analysis of selective binding of Au nanoparticles (NPs) to unblocked defects emphasized the superior repair mechanism in the branched layers with respect to the linear ones.

Journal Article↗

Sensitivity of transmission surface plasmon resonance (T-SPR) spectroscopy: self-assembled multilayers on evaporated gold island films.

The distance dependence of the localized surface plasmon (SP) extinction of discontinuous gold films is a crucial issue in the application of transmission surface plasmon resonance (T-SPR) spectroscopy to chemical and biological sensing. This derives from the usual sensing configuration, whereby an analyte binds to a selective receptor layer on the gold film at a certain distance from the metal surface. In the present work the distance sensitivity of T-SPR spectroscopy of 1.0-5.0 nm (nominal thickness) gold island films evaporated on silanized glass substrates is studied by using coordination-based self-assembled multilayers, offering thickness tuning in the range from approximately 1 to approximately 15 nm. The morphology, composition and optical properties of the Au/multilayer systems were studied at each step of multilayer construction. High-resolution scanning electron microscopy (HRSEM) showed no apparent change in the underlying Au islands, while atomic force microscopy (AFM) indicated flattening of the surface topography during multilayer construction. A regular growth mode of the organic layers was substantiated by X-ray photoelectron spectroscopy (XPS). Transmission UV-visible spectra showed an increase of the extinction and a red shift of the maximum of the SP band upon addition of organic layers, establishing the distance dependence of the Au SP absorbance. The distance sensitivity of T-SPR spectroscopy can be varied by using characteristic substrate parameters, that is, Au nominal thickness and annealing. In particular, effective sensitivity up to a distance of at least 15 nm is demonstrated with 5 nm annealed Au films. It is shown that intensity measurements, particularly in the plasmon intensity change (PIC) presentation, provide an alternative to the usually measured plasmon band position, offering good accuracy and the possibility of measuring at a single wavelength. The present distance sensitivity results provide the basis for further development of T-SPR transducers based on receptor-coated Au island films.

Journal Article↗

Coordination-based gold nanoparticle layers.

Gold nanoparticle (NP) mono- and multilayers were constructed on gold surfaces using coordination chemistry. Hydrophilic Au NPs (6.4 nm average core diameter), capped with a monolayer of 6-mercaptohexanol, were modified by partial substitution of bishydroxamic acid disulfide ligand molecules into their capping layer. A monolayer of the ligand-modified Au NPs was assembled via coordination with Zr4+ ions onto a semitransparent Au substrate (15 nm Au, evaporated on silanized glass and annealed) precoated with a self-assembled monolayer of the bishydroxamate disulfide ligand. Layer-by-layer construction of NP multilayers was achieved by alternate binding of Zr4+ ions and ligand-modified NPs onto the first NP layer. Characterization by atomic force microscopy (AFM), ellipsometry, wettability, transmission UV-vis spectroscopy, and cross-sectional transmission electron microscopy showed regular growth of NP layers, with a similar NP density in successive layers and gradually increased roughness. The use of coordination chemistry enables convenient step-by-step assembly of different ligand-possessing components to obtain elaborate structures. This is demonstrated by introducing nanometer-scale vertical spacing between a NP layer and the gold surface, using a coordination-based organic multilayer. Electrical characterization of the NP films was carried out using conductive AFM, emphasizing the barrier properties of the organic spacer multilayer. The results exhibit the potential of coordination self-assembly in achieving highly controlled composite nanostructures comprising molecules, NPs, and other ligand-derivatized components.

Gold↗

Preparative manipulation of gold nanoparticles by reversible binding to a polymeric solid support.

A preparative scheme is presented for controlled modification of gold nanoparticles (NPs) by using reversible binding to a polymeric solid support through boronic acid chemistry. Octanethiol-capped Au NPs were bound to a boronic acid functionalized resin by custom-synthesized bifunctional linker molecules. The NPs were chemically released from the resin to the solution, with one (or a few) linker molecules embedded in their capping layer. This was confirmed by rebinding the linker-derivatized NPs to a boronic resin, exploiting the reversibility of the boronic acid/diol chemistry. The same scheme was employed to demonstrate a new method for affinity separation of NPs by means of a solid-phase reaction. The use of boronic acid provides versatility and chemical reversibility, while the polymeric solid support affords the separation and preparative aspects. The method presented here may be useful in various facets of NP handling, manipulation, and separation.

Journal Article↗

Treatment of chronic rhinosinusitis with low-dose, long-term macrolide antibiotics: an evolving paradigm.

The 14-membered and 15-membered ring macrolide antibiotics express immunomodulatory effects in chronic respiratory disorders in humans that are distinct from their antimicrobial properties. These drugs downregulate the excessive immune and inflammatory responses observed in these conditions while promoting tissue repair. To this end, chronic rhinosinusitis is characterized by mucosal inflammation of nasal and sinus mucosa for more than 3 months and accounts for significant health care resource allocation due to difficulties in treatment. Clinical efficacy of macrolide antibiotics as biologic response modifiers in patients with chronic rhinosinusitis is suggested by compelling basic research and small, uncontrolled clinical studies. Hence, long-term, prospective double-blind placebo-controlled clinical studies are indicated to establish the utility of these drugs in the treatment of patients with chronic rhinosinusitis.

Animals↗

All D-VIP mitigates vasodilation elicited by L-VIP, micellar L-VIP and micellar PACAP1-38, but not PACAP1-38, in vivo.

The purpose of this study was to determine whether all D-vasoactive intestinal peptide (VIP), an inactive optical isomer of L-VIP, modulates the vasorelaxant effects of human L-VIP and pituitary adenylate cyclase activating peptide (PACAP)1-38, two ubiquitous and pleiotropic neuropeptides that activate VPAC1 and VPAC2, two VIP subtype receptors, in the intact peripheral microcirculation. Using intravital microscopy, we found that suffusion of all D-VIP had no significant effects on arteriolar diameter in the intact hamster cheek pouch. However, all D-VIP significantly attenuated L-VIP-induced vasodilation in a concentration-dependent fashion (P<0.05). likewise, all D-VIP significantly attenuated the vasorelaxant effects of L-VIP associated with sterically stabilized phospholipid micelles (SSM; P<0.05). Although all D-VIP had no significant effects on L-PACAP1-38-induced vasodilation, it abrogated PACAP1-38 in SSM-induced responses (P<0.05). The effects of all D-VIP were specific because it had no significant effects on acetylcholine-, nitroglycerin- and bradykinin-induced vasodilation. Taken together, these data indicate that all D-VIP attenuates the vasorelaxant effects of random coil and alpha-helix L-VIP as well as those of alpha-helix but not random coil PACAP in the intact peripheral microcirculation in a specific fashion. These effects are mediated, most likely, through interactions with VPAC1/VPAC2 receptors. We suggest that all D-VIP could be exploited as a novel, safe and active targeting moiety of VPAC1/VPAC2 receptors in vivo.

Acetylcholine↗

Liposomal vasoactive intestinal peptide.

Liposomes have been investigated as drug carriers since first discovered in the 1960s. However, the first-generation, so-called classic liposomes found relatively limited therapeutic utility. Nonetheless, the advent in the 1980s of the second-generation sterically stabilized liposomes (SSL) that evade uptake by the host's reticuloendothelial system greatly enhanced their utility as drug carriers because of their prolonged circulation half-life and passive targeting to injured and cancerous tissues. Over the past decade, our work focused on exploiting the bioactivity of vasoactive intestinal peptide (VIP), a ubiquitous 28-amino acid, amphipathic and pleiotropic mammalian neuropeptide, as a drug. To this end, the peptide expresses distinct and unique innate bioactivity that could be harnessed to treat several human diseases that represent unmet medical needs, such as pulmonary hypertension, stroke, Alzheimer's disease, sepsis, female sexual arousal dysfunction, acute lung injury, and arthritis. Unfortunately, the bioactive effects of VIP last only a few minutes due to its rapid degradation and inactivation by enzymes, catalytic antibodies, and spontaneous hydrolysis in biological fluids. Hence, our goal was to develop and test stable, long-acting formulations of VIP using both classic and SSL as platform technologies. We found that spontaneous association of VIP with phospholipid bilayers leads to a transition in the conformation of the peptide from random coil in an aqueous environment to alpha-helix, the preferred conformation for ligand-receptor interactions, in the presence of lipids. This process, in turn, protects VIP from degradation and inactivation and amplifies its bioactivity in vivo. Importantly, we discovered that the film rehydration and extrusion technique is the most suitable to passively load VIP onto SSL at room temperature and yields the most consistent results. Collectively, these attributes indicate that VIP on SSL represents a suitable formulation that could be tested in human disease.

Animals↗

Human VIP-alpha: an emerging biologic response modifier to treat primary pulmonary hypertension.

Primary pulmonary hypertension (PPH) is a rare life-threatening disorder of unknown etiology manifested by chronic elevation of pulmonary arterial pressure. Given that pulmonary vasoconstriction, endothelial and vascular smooth muscle cell proliferation and in situ thrombosis contribute appreciably to the evolution of PPH, treatment with vasodilators, antiproliferative drugs and anticoagulants, alone or in combination, constitute the pharmacologic standard of care. To this end, long-term administration of oral calcium channel blockers, prostacyclin analogs by various routes and oral endothelin-1 receptor antagonists, alone or in combination, is efficacious in treating patients with PPH. Unfortunately, efficacy is hampered by poor stability, delivery and bioavailability, and by systemic toxicity. Hence, there is an ongoing need to develop and test new drugs to treat patients with PPH. To address this issue, a novel, targeted, long-acting, biocompatible and safe sterically stabilized liposomal and micellar formulation of human vasoactive intestinal peptide (VIP) was developed and tested for human use: the 28-amino acid pleiotropic biologic response modifier, human VIP-alpha. The long-lasting salutary effects of phospholipid-associated VIP on vasomotor tone and arterial pressure were expressed at low concentrations solely in diseased animals and were independent of its route of administration. Thus, the author proposes that human VIP-alpha could be developed as a safe long-acting drug to treat patients with PPH.

Humans↗

Layer-by-layer assembly of ordinary and composite coordination multilayers.

Coordination self-assembly of bishydroxamate-based metal-organic multilayers on gold employing a layer-by-layer (LbL) approach was investigated. It is shown that the solution chemistry of the participating metal ion has a marked influence on the composition and properties of the multilayers. Use of Ce4+ and particularly zirconium(IV) acetylacetonate (Zr(acac)4) solutions in the ion-binding step of multilayer construction leads to multilayers with a near-stoichiometric metal ion-to-ligand ratio, suggesting a structure close to that predicted by a simple coordination self-assembly scheme. On the other hand use of a ZrCl4 solution as the source of metal ions in the multilayer construction leads to a multilayer with greater thickness and a large excess of Zr(IV), evenly distributed between the organic layers. In the latter case, a ratio of ca. 1:2 between the excess Zr and oxygen, as well as long-term Zr4+ binding experiments showing deposition of ZrO2, suggest the formation of a zirconia-type nanophase between the bishydroxamate organic repeat units during multilayer self-assembly. Hence, while the multilayer prepared using Zr(acac)4 solution appears to represent a "true" coordination-based structure, the one prepared using ZrCl4 is best described as a composite organic-ceramic multilayer. Composite multilayers prepared in this way display different properties from those of the stoichiometric ones, such as improved dielectric behavior and higher stiffness. Even greater mechanical stability is obtained with multilayers constructed using alternate binding of ZrCl4 and Ce4+. The concept of LbL formation of coordination-based composite organic-ceramic structures may be useful in obtaining nanometer-scale structures with tunable properties.

Cesium↗

Biological sensing using transmission surface plasmon resonance spectroscopy.

Ultrathin gold island films evaporated on transparent substrates offer promising transducers for chemical and biological sensing in the transmission surface plasmon resonance (T-SPR) mode. In the present work, the applicability of T-SPR-based systems to biosensing is demonstrated, using a well-established biological model system. Au island films were evaporated on polystyrene slides and modified with a biotinylated monolayer via a multistep surface reaction, the latter assisted by the good adhesion of metal islands to polystyrene. The biotin-derivatized Au island film was then used as a biological recognition surface for selective sensing of avidin binding, distinguishing between specific and nonspecific binding to the substrate. Transduction of the binding event into an optical signal was achieved by T-SPR spectroscopy, using plasmon intensity measurements, rather than wavelength change, for maximal sensitivity and convenience. T-SPR spectroscopy of Au island films is shown to be an effective tool for monitoring the binding of biological molecules to receptor layers on the Au surface and a promising approach to label-free optical biosensing.

Biosensing Techniques↗

Widely-applicable gold substrate for the study of ultrathin overlayers.

Ultrathin films on gold substrates have been the subject of enormous scientific and technological interest. Comprehensive study of such systems requires concomitant application of a variety of complementary characterization techniques. The reliability of the result is frequently hampered by the fact that different characterization methods impose different requirements on the Au substrate, resulting in the need to use different types of Au substrates for different measurements, possibly influencing the overlayer structure. This results in an average, rather than exact, structure determination. Here, we show that 15-nm-thick Au films evaporated at 0.5 A/sec on silanized glass and annealed are semi-transparent, electrically conducting, and morphologically well-defined, showing a smooth, [111] textured surface. Such Au films provide a high-quality, widely applicable and relatively inexpensive platform for ultrathin overlayers, enabling characterization by a wide spectrum of experimental methods, applied to the same substrate. The exceptional qualities and analytical capabilities of such substrates are demonstrated with several different systems: (i) Cu underpotential deposition (upd); (ii) alkanethiol self-assembly; (iii) formation of Au nanoparticle layers; (iv) binding of the chromophore protoporphyrin IX (PPIX) to a monolayer of 11-mercaptoundecanoic acid (MUA). In the latter case it is shown that the use of Cu(2+) ions for binding between the carboxylate groups of PPIX and MUA promotes better organization of the porphyrin layer.

Glass↗

Low-dose, long-term macrolide therapy in asthma: An overview.

Macrolides, a class of antimicrobials isolated from Streptomycetes more than 50 years ago, are used extensively to treat sinopulmonary infections in humans. In addition, a growing body of experimental and clinical evidence indicates that long-term (years), low (sub-antimicrobial)-dose 14- and 15-membered ring macrolide antibiotics, such as erythromycin, clarithromycin, roxithromycin and azithromycin, express immunomodulatory and tissue reparative effects that are distinct from their anti-infective properties. These salutary effects are operative in various lung disorders, including diffuse panbronchiolitis, cystic fibrosis, persistent chronic rhinosinusitis, nasal polyposis, bronchiectasis, asthma and cryptogenic organizing pneumonia.The purpose of this overview is to outline the immunomodulatory effects of macrolide antibiotics in patients with asthma.

Journal Article↗

In vitro characterization of PEGylated phospholipid micelles for improved drug solubilization: effects of PEG chain length and PC incorporation.

Sterically stabilized micelles (SSM) composed of poly(ethylene glycol-2000)-grafted distearoylphosphatidylethanolamine (DSPE-PEG) and sterically stabilized mixed micelles (SSMM) composed of DSPE-PEG and egg-phosphatidyl choline (PC) have recently been introduced as novel lipid based carriers for water-insoluble drugs. However, factors that affect the solubilization behavior of these phospholipid micelles are not well understood. This study investigates the effect of PEG chain length and PC content on physical properties and solubilization potential of PEGylated phospholipid micelles. Critical micelle concentrations (CMC) determined for DSPE-PEG with different PEG chain lengths (2000, 3000, and 5000) using a fluorescent probe were in the micromolar range (0.5-1.5 microM) with higher CMC for longer PEG chain length. The size of micelles determined by quasi-elastic light scattering (QELS) showed that micellar systems became heterogeneous when PC was added at > or =25% for DSPE-PEG 2000 and > or = 40% for DSPE-PEG 5000, respectively. Above these critical PC ratios a significant increase in aggregation number and formation of rodlike particles were observed by small angle neutron scattering (SANS). Solubilization of diazepam was greater with DSPE-PEG 2000 than DSPE-PEG 5000 simple micelles as determined by RP-HPLC. However, DSPE-PEG 5000 micelles showed greater improvement in solubilization of the water-insoluble drug with an increase in PC content. In conclusion, phospholipid micelle size and solubilization potential varied with PEG chain length and PC content in the mixed micelle. Aggregation number and shape of the micelles did not significantly change until the critical PC concentrations.

Diazepam↗

Helospectin I and II evoke vasodilation in the intact peripheral microcirculation.

Helospectin I and II, two closely related mammalian neuropeptides of the secretin/glucagons/vasoactive intestinal peptide (VIP) superfamily of peptides, are co-localized with VIP in nerve fibers surrounding vascular smooth muscle. However, the role if any, VIP receptors play in transducing the vasorelaxant effects of helospectin I and II in the intact peripheral microcirculation is uncertain. The purpose of this study was to determine whether helospectin I and II elicit vasodilation in the intact peripheral microcirculation and, if so, whether this response is mediated, in part, by VIP or pituitary adenylate cyclase activating peptide (PACAP) receptor engagement, and through local elaboration of cyclooxygenase products of arachidonic acid metabolism. Using intravital microscopy, we found that suffusion of helospectin I and II (each, 1.0 nmol) evoked potent vasodilation and of similar magnitude in the intact hamster cheek pouch microcirculation (P < 0.05). Suffusion of 0.1 nmol helospectin I and II had no significant effects on arteriolar diameter. Pretreatment with VIP(10-28), a VPAC1/VPAC2 receptor antagonist, or PACAP(6-38), a PAC1/VPAC2 receptor antagonist, had no significant effects on helospectin I- and II-induced responses. In addition, pretreatment with indomethacin had no significant effects on helospectin I- and II-induced vasodilation. Collectively, these data indicate that helospectin I and II evoke potent vasodilation in the intact peripheral microcirculation that is not transduced by VIP or PACAP receptors nor through cyclooxygenase products of arachidonic acid metabolism.

Animals↗