Analysis of biotinylated generation 4 poly(amidoamine) (PAMAM) dendrimer distribution in the rat brain and toxicity in a cellular model of the blood-brain barrier.

Dendrimers are highly customizable nanopolymers with qualities that make them ideal for drug delivery. The high binding affinity of biotin/avidin provides a useful approach to fluorescently label synthesized dendrimer-conjugates in cells and tissues. In addition, biotin may facilitate delivery of dendrimers through the blood-brain barrier (BBB) via carrier-mediated endocytosis. The purpose of this research was to: (1) measure toxicity using lactate dehydrogenase (LDH) assays of generation (G)4 biotinylated and non-biotinylated poly(amidoamine) (PAMAM) dendrimers in a co-culture model of the BBB, (2) determine distribution of dendrimers in the rat brain, kidney, and liver following systemic administration of dendrimers, and (3) conduct atomic force microscopy (AFM) on rat brain sections following systemic administration of dendrimers. LDH measurements showed that biotinylated dendrimers were toxic to cell co-culture after 48 h of treatment. Distribution studies showed evidence of biotinylated and non-biotinylated PAMAM dendrimers in brain. AFM studies showed evidence of dendrimers only in brain tissue of treated rats. These results indicate that biotinylation does not decrease toxicity associated with PAMAM dendrimers and that biotinylated PAMAM dendrimers distribute in the brain. Furthermore, this article provides evidence of nanoparticles in brain tissue following systemic administration of nanoparticles supported by both fluorescence microscopy and AFM.

Giant magnetoresistance sensors. 1. Internally calibrated readout of scanned magnetic arrays.

This paper describes efforts aimed at setting the stage for the application of giant magnetoresistance sensor (GMRs) networks as readers for quantification of biolytes selectively captured and then labeled with superparamagnetic particles on a scanned chip-scale array. The novelty and long-range goal of this research draws from the potential development of a card-swipe instrument through which an array of micrometer-sized, magnetically tagged addresses (i.e., a sample stick) can be interrogated in a manner analogous to a credit card reader. This work describes the construction and testing of a first-generation instrument that uses a GMR sensor network to read the response of a "simulated" sample stick. The glass sample stick is composed of 20-nm-thick films of permalloy that have square or rectangular lateral footprints of up to a few hundred micrometers. Experiments were carried out to gain a fundamental understanding of the dependence of the GMR response on the separation between, and planarity of, the scanned sample stick and sensor. Results showed that the complex interplay between these experimentally controllable variables strongly affect the shape and magnitude of the observed signal and, ultimately, the limit of detection. This study also assessed the merits of using on-sample standards as internal references as a facile means to account for small variations in the gap between the sample stick and sensor. These findings were then analyzed to determine various analytical figures of merit (e.g., limit of detection in terms of the amount of magnetizable material on each address) for this readout strategy. An in-depth description of the first-generation test equipment is presented, along with a brief discussion of the potential widespread applicability of the concept.

Giant magenetoresistive sensors. 2. Detection of biorecognition events at self-referencing and magnetically tagged arrays.

Microfabricated devices formed from alternating layers of magnetic and nonmagnetic materials at combined thicknesses of a few hundred nanometers exhibit a phenomenon known as the giant magnetoresistance effect. Devices based on this effect are known as giant magnetoresistive (GMR) sensors. The resistance of a GMR is dependent on the strength of an external magnetic field, which has resulted in the widespread usage of such platforms in high-speed, high-data density storage drives. The same attributes (i.e., sensitivity, small size, and speed) are also important embodiments of many types of bioanalytical sensors, pointing to an intriguing opportunity via an integration of GMR technology, magnetic labeling strategies, and biorecognition elements (e.g., antibodies). This paper describes the utilization of GMRs for the detection of streptavidin-coated magnetic particles that are selectively captured by biotinylated gold addresses on a 2 x 0.3 cm sample stick. A GMR sensor network reads the addresses on a sample stick in a manner that begins to emulate that of a "card-swipe" system. This study also takes advantage of on-sample magnetic addresses that function as references for internal calibration of the GMR response and as a facile means to account for small variations in the gap between the sample stick and sensor. The magnetic particle surface coverage at the limit of detection was determined to be approximately 2%, which corresponds to approximately 800 binding events over the 200 x 200 microm capture address. These findings, along with the potential use of streptavidin-coated magnetic particles as a universal label for antigen detection in, for example, heterogeneous assays, are discussed.

Use of attenuated total reflectance Fourier transform infrared spectroscopy to identify microbial metabolic products on carbonate mineral surfaces.

This paper demonstrates the use of attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy to detect microbial metabolic products on carbonate mineral surfaces. By creating an ATR-FTIR spectral database for specific organic acids using ATR-FTIR spectroscopy we were able to distinguish metabolic acids on calcite surfaces following Escherichia coli growth. The production of these acids by E. coli was verified using high-performance liquid chromatography with refractive index detection. The development of this technique has allowed us to identify microbial metabolic products on carbonate surfaces in nutrient-limited cave environments.

Low-Level Detection of Poly(amidoamine) PAMAM Dendrimers Using Immunoimaging Scanning Probe Microscopy.

Immunoimaging scanning probe microscopy was utilized for the low-level detection and quantification of biotinylated G4 poly(amidoamine) PAMAM dendrimers. Results were compared to those of high-performance liquid chromatography (HPLC) and found to provide a vastly improved analytical method for the low-level detection of dendrimers, improving the limit of detection by a factor of 1000 (LOD = 2.5 × 10(-13) moles). The biorecognition method is reproducible and shows high specificity and good accuracy. In addition, the capture assay platform shows a promising approach to patterning dendrimers for nanotechnology applications.

Surface complexation of catechol to metal oxides: an ATR-FTIR, adsorption, and dissolution study.

The interaction of catechol with chromium(III) oxide (Cr(2)O(3)), manganese dioxide (MnO(2)), iron(III) oxide (Fe(2)O(3)), and titanium dioxide (TiO(2)) was evaluated as a function of pH conditions (pH 3-10) and ionic strength using a combined approach of bulk adsorption, attenuated total reflectance Fourier-transform infrared spectroscopy (ATR-FTIR), and dissolution analysis. Adsorption of catechol showed a strong pH-dependent behavior with the metal oxides, remaining constant under acidic-neutral pH (3-7) and increasing under more basic conditions. In situ ATR-FTIR measurements indicate that catechol binds predominately as an outer-sphere complex on MnO(2) and as an inner-sphere complex on Fe(2)O(3), TiO(2), and Cr(2)O(3) substrates. Catechol complexation on Fe(2)O(3), TiO(2), and Cr(2)O(3) promotes dissolution at pH >5, whereas MnO(2) dissolution occurs under acidic and basic conditions (pH 3-10).