Magnetically controlled release of cisplatin from superparamagnetic starch nanoparticles

The present study involves a novel strategy for the preparation of superparamagnetic nanoparticles of crosslinked starch impregnated with homogeneously dispersed nanosized iron oxide. The nanoparticles were loaded with an anticancer drug ‘cisplatin’ and the drug release kinetics was investigated spectrophotometrically at physiological pH (7.4). The nanoparticles were characterized by Fourier transform infra red (FTIR) spectroscopy, transmission electron microscopy (TEM), X-ray diffraction and magnetization studies. The particle size of magnetic starch nanoparticles was found to lie in the range of 20-90 nm. The influence of factors like chemical composition of nanoparticles, pH and temperature of the release media and applied magnetic field was investigated on the release profiles of the drug. The prepared nanoparticles could provide a possible pathway for targeted and controlled delivery of anticancer drugs minimizing side effects and achieving higher efficacy.     

Mitotic cell populations obtained from a micro-carrier culturing system

Colcemid treatment of cell culture beads confluent with cells released a mitotic population of cells which was as efficient as that obtained by conventional shake-off procedures. The bead system provides an easy and efficient means for harvesting large quantities of mitotic cells, costs about one-tenth as much as conventional culturing techniques, and is at least 100 times more rapid.     

Microparticulate systems for targeted drug delivery to phagocytes

Comment on: Jhunjhunwala S, et al. J Control Release 2009;133:191-7.     

A photochemical approach for controlled drug release in targeted drug delivery

Photochemistry provides a unique mechanism that enables the active control of drug release in cancer-targeting drug delivery. This study investigates the light-mediated release of methotrexate, an anticancer drug, using a photocleavable linker strategy based on o-nitrobenzyl protection. We evaluated two types of the o-nitrobenzyl-linked methotrexate for the drug release study and further extended the study to a fifth-generation poly(amidoamine) dendrimer carrier covalently conjugated with methotrexate via the o-nitrobenzyl linker. We performed the drug release studies by using a combination of three standard analytical methods that include UV/vis spectrometry, (1)H NMR spectroscopy, and anal. HPLC. This article reports that methotrexate is released by the photochemical mechanism in an actively controlled manner. The rate of the drug release varies in response to multiple control parameters, including linker design, light wavelength, exposure time, and the pH of the medium where the drug release occurs.     

Eicosanomics: targeted lipidomics of eicosanoids in biological systems

Recent advancements in mass spectrometry, especially the development of electrospray tandem mass spectrometry (ESI/LC/MS2) and matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI/TOF), have greatly facilitated analysis of complex biomolecules. It has now become possible to profile, in relatively short periods of time, large multicomponent groups of compounds biosynthesized by biological systems. The efficiency and accuracy of analysis have led to the development of new concepts of mass spectrometric profiling, mapping, and imaging. Profiling of proteins in biological material (proteomics) has become a widely accepted strategy for identification of mechanisms involved in the biochemistry of disease processes, and has become a novel tool for unraveling new drug targets. Evolution of proteomics has relied on ESI/LC/MS2 and MALDI/TOF, techniques that are also useful in the novel area of quantitative proteomics.     

Devices and systems targeted towards augmented robotic radical prostatectomy

Prostate cancer is the most frequent male cancer and the second cause of male cancer mortality in developed countries. Therefore, it represents a major public health issue. Health problem and the development of new therapeutic strategies to address this issue is essential. During a prostatectomy, the surgeon looks for a compromise between an exhaustive removal of pathologic tissue (to achieve the best carcinogenic prognosis) and the functional consequences linked to a wide excision (i.e.: avoid as much as possible urinary incontinence and sexual dysfunction). In this context, the ANR TecSan DEPORRA project regroups French research laboratories (TIMC-IMAG, FEMTO-ST), companies (Endocontrol-Medical, VERMON) and hospital departments (CIC-IT, Urology & pathology Department of the Grenoble University Hospital) to bring innovative tools for radical prostatectomy. These tools will provide to the surgeon new information from several imaging modalities (video, fluorescence and US imaging), and combine them in an augmented environment. We believe that this augmented environment will ultimately help the surgeon to perform his surgical gesture “optimally” and will improve the patient's carcinogenic and functional prognosis.     

Two systems for targeted gene deletion in Coxiella burnetii

Coxiella burnetii is a ubiquitous zoonotic bacterial pathogen and the cause of human acute Q fever, a disabling influenza-like illness. C. burnetii's former obligate intracellular nature significantly impeded the genetic characterization of putative virulence factors. However, recent host cell-free (axenic) growth of the organism has enabled development of shuttle vector, transposon, and inducible gene expression technologies, with targeted gene inactivation remaining an important challenge. In the present study, we describe two methods for generating targeted gene deletions in C. burnetii that exploit pUC/ColE1 ori-based suicide plasmids encoding sacB for positive selection of mutants. As proof of concept, C. burnetii dotA and dotB, encoding structural components of the type IVB secretion system (T4BSS), were selected for deletion. The first method exploited Cre-lox-mediated recombination. Two suicide plasmids carrying different antibiotic resistance markers and a loxP site were integrated into 5' and 3' flanking regions of dotA. Transformation of this strain with a third suicide plasmid encoding Cre recombinase resulted in the deletion of dotA under sucrose counterselection. The second method utilized a loop-in/loop-out strategy to delete dotA and dotB. A single suicide plasmid was first integrated into 5' or 3' target gene flanking regions. Resolution of the plasmid cointegrant by a second crossover event under sucrose counterselection resulted in gene deletion that was confirmed by PCR and Southern blot. ΔdotA and ΔdotB mutants failed to secrete T4BSS substrates and to productively infect host cells. The repertoire of C. burnetii genetic tools now allows ready fulfillment of molecular Koch's postulates for suspected virulence genes.     

Magnetically labeled insulin-secreting cells

Iron oxide nanoparticles have been shown to magnetically label cells in order to visualize them in vivo via MR imaging. This technology has yet to be implemented in insulin secreting cells, thus it is not known whether the presence of these nanoparticles in the cytoplasm of the cells affects insulin secretion. This study investigates the effectiveness and consequence of labeling mouse insulinoma betaTC3 and betaTC-tet cells with monocrystalline iron oxide nanoparticles (MION). Our data show that MION can be internalized in both betaTC3 and betaTC-tet cells following a 24h exposure to 0.02mg/ml MION solution. The metabolic and secretory activities of both MION-labeled cell lines were statistically indistinguishable from sham treatment. Furthermore, cell viability and apoptosis remained constant throughout the cell's exposure to MION. Finally, MR images demonstrated significant contrast between labeled and sham-treated cells. Thus, labeling murine insulinoma cell lines with magnetic iron oxide nanoparticles does not hinder their insulin secretion, while it provides MR imaging contrast.     

Regulation of cell polarity by controlled proteolytic systems

Epithelial and neuronal cells are highly asymmetric, with discrete regions responsible for different roles that underlie the generation of specific compartments within cells that are distinct in biochemical composition, structure, and morphology that ultimately lead to distinct functions. Controlled and specific molecular targeting and sorting have been studied to understand the generation of asymmetric domains inside cells. Recently, a new and complementary explanation has emerged to account for the generation of domains that are enriched by a subset of proteins or polarization determinants: local proteolysis. In this review, we discuss the most conspicuous proteolytic systems that may contribute to the generation of cell polarity, namely the ubiquitin-proteosome and the calpain systems. Specifically, we focus this review on two cellular processes that depend on the acquisition of cell polarity; cell migration and the establishment of an axon in a neuronal cell.     

Synthesis of GnRH analogs and their application in targeted gene delivery systems

A set of GnRH analogues containing nuclear localization signal (NLS) of SV-40 virus large T-antigen have been synthesized using solid phase peptide synthesis and chemical ligation technique. Selective chemical ligation was achieved as a result of hydrazone formation in the course of interaction between NLS hydrazide and GnRH analog modified by pyruvic acid. The efficiency of synthesized peptide carriers was demonstrated in experiments with human cancer cells transfected by reporter luciferase and beta-galactosidase genes or suicide HSV-1 thymidine kinase gene. It was shown that selectivity of action on cancer cells can be achieved as a result of peptide/DNA complex penetration through the cell membrane by GnRH receptor-mediated endocytosis pathway.     

Synthesis of GnRH analogues and their application in targeted gene delivery systems

A set of GnRH analogues containing the nuclear localization signal (NLS) of the SV-40 virus was synthesized using solid phase peptide synthesis and chemical ligation techniques. Selective chemical ligation was achieved through hydrazone formation upon the interaction of NLS hydrazide and GnRH analogue modified with pyruvic acid. The efficiency of the synthesized compounds was demonstrated in experiments on transfection of various human cancer cell lines with reporter luciferase and β-galactosidase genes, as well as suicide thymidine kinase gene of HSV-1. Selectivity of the peptide-DNA complex effect on cancer cells is achieved as a result of its penetration through the cell membrane via GnRH receptor-mediated endocytosis pathway.     

Responsive polymer systems for controlled delivery of therapeutics.

The ideal drug-delivery system should provide therapeutics in response to physiological requirements, having the capacity to 'sense' changes and alter the drug-release process accordingly. Such responsive controlled delivery systems are still at an experimental stage. This review focuses on two basic approaches: (1) externally regulated systems (utilizing triggers such as magnetism, ultrasound, temperature and electricity), and (2) self-regulated systems (utilizing pH-sensitive polymers, enzyme-substrate reactions, competitive binding, and antibody interactions).