Synthesis, spectral and quantum chemical studies on NO-chelating sulfamonomethoxine-cyclophosph(V)azane and its Er(III) complex.

Computational studies have been carried out at the DFT-B3LYP/6-31G(d) level of theory on the structural and spectroscopic properties of novel ethane-1,2-diol-dichlorocyclophosph(V)azane of sulfamonomethoxine (L), and its binuclear Er(III) complex. Different tautomers of the ligand were optimized at the ab initio DFT level. Keto-form structure is about 15.8 kcal/mol more stable than the enol form (taking zpe correction into account). Simulated IR frequencies were scaled and compared with that experimentally measured. TD-DFT method was used to compute the UV-VIS spectra which show good agreement with measured electronic spectra. The structures of the novel isolated products are proposed based on elemental analyses, IR, UV-VIS, (1)H NMR, (31)P NMR, SEM, XRD spectra, effective magnetic susceptibility measurements and thermogravimetric analysis (TGA).

Using fluorescence correlation spectroscopy (FCS) for IFN-g detection: a preliminary study.

Nowadays, enzyme-linked immunosorbent assay (ELISA) based detection of Mycobacterium tuberculosis (M. tuberculosis) antigen triggered interferon-gamma (IFN-g) secretion by blood T cells displays an improved diagnostic value for M. tuberculosis infection. Applications of fluorescence correlation spectroscopy (FCS) have been explored in various subfields of medicine and molecular biology, including detection of a certain biomarker in liquid instead of ELISA. Here, we present a preliminary study of detecting IFN-g using FCS-based technique.

Tumor targeting and imaging with dual-peptide conjugated multifunctional liposomal nanoparticles.

BACKGROUND: The significant progress in nanotechnology provides a wide spectrum of nanosized material for various applications, including tumor targeting and molecular imaging. The aim of this study was to evaluate multifunctional liposomal nanoparticles for targeting approaches and detection of tumors using different imaging modalities. The concept of dual-targeting was tested in vitro and in vivo using liposomes derivatized with an arginine-glycine-aspartic acid (RGD) peptide binding to αvß3 integrin receptors and a substance P peptide binding to neurokinin-1 receptors. METHODS: For liposome preparation, lipids, polyethylene glycol building blocks, DTPA-derivatized lipids for radiolabeling, lipid-based RGD and substance P building blocks and imaging labels were combined in defined molar ratios. Liposomes were characterized by photon correlation spectroscopy and zeta potential measurements, and in vitro binding properties were tested using fluorescence microscopy. Standardized protocols for radiolabeling were developed to perform biodistribution and micro-single photon emission computed tomography/computed tomography (SPECT/CT) studies in nude mice bearing glioblastoma and/or melanoma tumor xenografts. Additionally, an initial magnetic resonance imaging study was performed. RESULTS: Liposomes were radiolabeled with high radiochemical yields. Fluorescence microscopy showed specific cellular interactions with RGD-liposomes and substance P-liposomes. Biodistribution and micro-SPECT/CT imaging of (111)In-labeled liposomal nanoparticles revealed low tumor uptake, but in a preliminary magnetic resonance imaging study with a single-targeted RGD-liposome, uptake in the tumor xenografts could be visualized. CONCLUSION: The present study shows the potential of liposomes as multifunctional targeted vehicles for imaging of tumors combining radioactive, fluorescent, and magnetic resonance signaling. Specific in vitro tumor targeting by fluorescence microscopy and radioactivity was achieved. However, biodistribution studies in an animal tumor model revealed only moderate tumor uptake and no additive effect using a dual-targeting approach.

Liposomal vasoactive intestinal peptide for lung application: protection from proteolytic degradation.

Inhalative administration of vasoactive intestinal peptide (VIP) is a promising approach for the treatment of severe lung diseases. However, the clinical use of VIP is limited by the fact that the peptide is prone to rapid degradation mechanisms and proteolytic digestion. Accordingly, VIP exhibits a very short period of activity in the lung. To overcome this problem, we have designed a liposomal drug delivery system for VIP and characterized it in terms of its potential to protect VIP from enzymatic cleavage. The proteolytic conditions of the lung, the target site of aerosolic administered VIP, were mimicked by bronchoalveolar lavage fluid (BALF), a lung surfactant solution, obtained by fiberoptic bronchoscopy. Thus, the stability of VIP was assessed by its resistance to enzymatic degradation in BALF, using a combination of high pressure liquid chromatography with mass spectrometry. We found that free VIP was rapidly digested, whereas liposomal-associated VIP remained intact. By fluorescence spectroscopic techniques using fluorescent-labelled VIP we got strong indications that the tight association of VIP with the lipid membrane is only minimally affected upon incubation with BALF. Loading capacity and stability of EtCy3-VIP loaded liposomes were measured by fluorescence fluctuation spectroscopy. Finally, the protective properties of the liposomes were also expressed in the maintained biological activity of the peptide incubated with BALF.

Monovalent ion dependence of neomycin B binding to an RNA aptamer characterized by spectroscopic methods.

Understanding the interactions of small molecules like antibiotics with RNA is a prerequisite for the development of novel drugs. In this study we address structural and thermodynamic features of such interactions by using a simple model system: the binding of the highly charged antibiotic neomycin B to a short hairpin RNA molecule. Nucleotide A16, which acts as a flap over the neomycin B binding pocket, was substituted by the fluorescent adenine analogue 2-aminopurine (2-AP). Steady-state and time-resolved fluorescence measurements were complemented by UV-melting and circular dichroism studies. The binding of neomycin B at three sites was found to have a strong inverse correlation with Na(+) concentration. For the highest-affinity site, both fluorescence and UV absorption experiments were consistent with a model assuming at least three neomycin NH(3) (+) groups participating in addition to hydrogen bonds in electrostatic interactions with the RNA. The variation of fluorescence intensity and lifetime upon neomycin B binding indicated unstacking of 2-AP16 from neighbouring bases as it flipped over the binding pocket. RNA conformational changes upon binding of the antibiotic were confirmed by circular dichroism. The two weaker binding sites were characterized as unspecific binding to the aptamer, while the high-affinity binding event was shown to be highly specific even at high ionic concentration. In addition, 2-AP was confirmed to be a noninvasive fluorescent probe; it serves as a sensitive spectroscopic tool to investigate details of the interactions between small molecules and RNA.