Use of Arabidopsis eceriferum Mutants to Explore Plant Cuticle Biosynthesis

JoVE 709 5/31/2008

Lacey Samuels¹, Allan DeBono¹, Patricia Lam¹, Miao Wen¹, Reinhard Jetter^1,2, Ljerka Kunst¹

¹Department of Botany, University of British Columbia - UBC, ²Department of Chemistry, University of British Columbia - UBC

The plant cuticle is a waxy outer covering on plants that has a primary role in water conservation but is also an important barrier against the entry of pathogenic microorganisms. In this video, we demonstrate the analysis of plant cuticle mutants identified by forward and reverse genetics approaches.

One-step Metabolomics: Carbohydrates, Organic and Amino Acids Quantified in a Single Procedure

JoVE 2014 6/25/2010

James D. Shoemaker

Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine

The urease method of sample preparation for GC/MS analysis of intermediary metabolites is presented by its inventor. The method allows one-step follow-up of newborn screening for inborn errors by tandem mass spectrometry by quantifying carbohydrates, organic and amino acids all in a single process.

A Toolkit to Enable Hydrocarbon Conversion in Aqueous Environments

JoVE 4182 10/02/2012

Eva K. Brinkman¹, Kira Schipper¹, Nadine Bongaerts¹, Mathias J. Voges¹, Alessandro Abate², S. Aljoscha Wahl¹

¹Department of Biotechnology, Delft University of Technology, ²Delft Center for Systems and Control, Delft University of Technology

A sustainable auto regulating bacterial system for the remediation of oil pollutions was designed using standard interchangeable DNA parts (BioBricks). An engineered E. coli strain was used to degrade alkanes via β-oxidation in toxic aqueous environments. The respective enzymes from different species showed alkane degradation activity. Additionally, an increased tolerance to n-hexane was achieved by introducing genes from alkane-tolerant bacteria.

Monitoring Plant Hormones During Stress Responses

JoVE 1127 6/15/2009

Marie J. Engelberth, Jurgen Engelberth

Department of Biology, University of Texas

A simple method is provided that allows for the rapid extraction and analysis of multiple plant hormones from small tissue samples. The procedure uses vapor phase extraction as the solemn purification step. Samples are analyzed by GC/MS with chemical ionization that produces mainly (M+1)+ ions.

Biochemical Measurement of Neonatal Hypoxia

JoVE 2948 8/24/2011

Megan S. Plank¹, Teleka C. Calderon¹, Yayesh Asmerom², Danilo S. Boskovic¹, Danilyn M. Angeles²

¹Division of Biochemistry, Department of Basic Sciences, Loma Linda University, ²Division of Physiology, Department of Basic Sciences, Loma Linda University

A method is described to measure biochemical markers of neonatal hypoxia-ischemia. The approach utilizes high pressure liquid chromatography (HPLC) and Gas Chromatography Mass Spectrometry (GC/MS).

Stable Isotopic Profiling of Intermediary Metabolic Flux in Developing and Adult Stage Caenorhabditis elegans

JoVE 2288 2/27/2011

Marni J. Falk^1,2, Meera Rao*¹, Julian Ostrovsky*¹, Evgueni Daikhin¹, Ilana Nissim¹, Marc Yudkoff^1,2

¹Department of Pediatrics, The Children's Hospital of Philadelphia, ²Department of Pediatrics, University of Pennsylvania

Stable isotopic profiling by gas chromatography mass spectrometric analysis of intermediary metabolic flux is described in the nematode, Caenorhabditis elegans. Methods are detailed for assessing isotopic enrichment in carbon dioxide, organic acids, and amino acids following isotope exposure either during development on agar plates or during adulthood in liquid culture.

Identification of Olfactory Volatiles using Gas Chromatography-Multi-unit Recordings (GCMR) in the Insect Antennal Lobe

JoVE 4381 2/24/2013

Kelsey J. R. P. Byers, Elischa Sanders, Jeffrey A. Riffell

Department of Biology, University of Washington

Olfactory cues mediate many different behaviors in insects, and are often complex mixtures comprised of tens to hundreds of volatile compounds. Using gas chromatography with multi-channel recording in the insect antennal lobe, we describe a method for the identification of bioactive compounds.

Biochemical and High Throughput Microscopic Assessment of Fat Mass in Caenorhabditis Elegans

JoVE 50180 3/30/2013

Elizabeth C. Pino¹, Christopher M. Webster¹, Christopher E. Carr², Alexander A. Soukas¹

¹Center for Human Genetic Research and Department of Medicine, Massachusetts General Hospital and Harvard Medical School, ²Department of Earth, Atmospheric, and Planetary Sciences, Massachusetts Institute of Technology

We present robust biochemical and microscopic methods for studying Caenorhabditis elegans lipid stores. A rapid, simple, fixing-staining procedure for fluorescent lipid droplet imaging leverages the spectral properties of the lipophilic dye Nile red. We then present biochemical measurement of triglycerides and phospholipids using solid phase extraction and gas chromatography-mass spectrometry.

Microdialysis of Ethanol During Operant Ethanol Self-administration and Ethanol Determination by Gas Chromatography

JoVE 4142 9/05/2012

Christina J. Schier, Regina A. Mangieri, Geoffrey A. Dilly, Rueben A. Gonzales

College of Pharmacy, Division of Pharmacology and Toxicology, The University of Texas at Austin

A method to determine the time course of ethanol concentration in the brains of rats during operant ethanol self-administration is described. Gas chromatography with flame ionization detection is used to quantify ethanol in the dialysate samples, because it has the sensitivity required for the small volumes that are generated.

GC-based Detection of Aldononitrile Acetate Derivatized Glucosamine and Muramic Acid for Microbial Residue Determination in Soil

JoVE 3767 5/19/2012

Chao Liang^1,2, Harry W. Read², Teri C. Balser^2,3

¹DOE-Great Lakes Bioenergy Research Center, University of Wisconsin, Madison, ²Department of Soil Science, University of Wisconsin, Madison, ³Department of Soil and Water Science, University of Florida

We describe a method protocol for the GC-based analysis of the aldonitrile acetate derivatives of glucosamine and muramic acid extracted from soil. For elucidation of the chemical mechanism, we also present a strategy to confirm the structure of the derivative and the ion fragments formed upon electron ionization.

Fruit Volatile Analysis Using an Electronic Nose

JoVE 3821 3/30/2012

Simona Vallone¹, Nathan W. Lloyd², Susan E. Ebeler³, Florence Zakharov¹

¹Department of Plant Sciences, University of California, Davis, ²Department of Chemical Engineering and Material Science, University of California, Davis, ³Department of Viticulture and Enology, University of California, Davis

A rapid method for volatile compound analysis in fruit is described. The volatile compounds present in the headspace of a homogenate of the sample are rapidly separated and detected with ultra-fast gas chromatography (GC) coupled with a surface acoustic wave (SAW) sensor. A procedure for data handling and analysis is also discussed.

March 2012: This Month in JoVE

JoVE 4336 3/01/2012

Aaron Kolski-Andreaco

Here are some highlights from the March 2012 Issue of Journal of Visualized Experiments (JoVE).

Mass Spectrometric Analysis of Glycosphingolipid Antigens

JoVE 4224 4/16/2013

Alexandra Bili Yin¹, David Hawke², Dapeng Zhou^3,4

¹Undergraduate Program, Rice University, ²Proteomics Facility, Department of Pathology, University of Texas MD Anderson Cancer Center, ³Department of Melanoma Medical Oncology, University of Texas MD Anderson Cancer Center, ⁴University of Texas Graduate School of Biological Sciences at Houston

A specific and sensitive method to gain insight into the expression profile of glycosphingolipid antigens in immune organs and cells is described. The method takes advantage of the ion trap mass spectrometry allowing step-wise fragmentation of glycosphingolipid molecules for structural analysis in comparison to chemically synthesized standards.

Free Radicals in Chemical Biology: from Chemical Behavior to Biomarker Development

JoVE 50379 4/15/2013

Chryssostomos Chatgilialoglu, Carla Ferreri, Annalisa Masi, Michele Melchiorre, Anna Sansone, Michael A. Terzidis, Armida Torreggiani

ISOF - Bio Free Radicals, Consiglio Nazionale delle Ricerche

Radical-based biomimetic chemistry has been applied to building-up libraries necessary for biomarker development.

A Quantitative Assessment of The Yeast Lipidome using Electrospray Ionization Mass Spectrometry

JoVE 1513 8/21/2009

Simon D. Bourque, Vladimir I. Titorenko

Department of Biology, Concordia University

We describe a new quantitative lipidomics method for identifying numerous lipid species in yeast using survey-scan electrospray ionization mass spectrometry (ESI/MS). This method exceeds currently available methods for lipid identification and quantification in the ability to resolve various molecular forms of lipids, sensitivity, and speed.

Metabolic Pathway Confirmation and Discovery Through ¹³C-labeling of Proteinogenic Amino Acids

JoVE 3583 1/26/2012

Le You¹, Lawrence Page², Xueyang Feng¹, Bert Berla¹, Himadri B. Pakrasi³, Yinjie J. Tang¹

¹Department of Energy, Environmental and Chemical Engineering, Washington University, ²Department of Biology, Washington University, ³Department of Energy, Environmental and Chemical Engineering and Department of Biology, Washington University

¹³C-isotope labeling is a useful technique for determining the cell central metabolism for various types of microorganisms. After cells have been cultured with a specific labeled substrate, GC-MS measurement can reveal functional metabolic pathways based on unique labeling patterns in proteinogenic amino acids.

Sample Preparation of Mycobacterium tuberculosis Extracts for Nuclear Magnetic Resonance Metabolomic Studies

JoVE 3673 9/03/2012

Denise K. Zinniel¹, Robert J. Fenton¹, Steven Halouska², Robert Powers², Raul G. Barletta¹

¹School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, ²Department of Chemistry, University of Nebraska-Lincoln

The metabolomic profile of Mycobacterium tuberculosis is determined after growth in broth cultures. Conditions can be varied to test the effects of nutritional supplements, oxidants, and anti-tuberculosis agents on the metabolic profile of this microorganism. Procedure for extract preparation is applicable for both 1D ¹H and 2D ¹H-¹³C NMR analyses.

Concentration of Metabolites from Low-density Planktonic Communities for Environmental Metabolomics using Nuclear Magnetic Resonance Spectroscopy

JoVE 3163 4/07/2012

R. Craig Everroad*¹, Seiji Yoshida*², Yuuri Tsuboi¹, Yasuhiro Date³, Jun Kikuchi^2,3,4, Shigeharu Moriya^1,2

¹Biosphere Oriented Biology Research Unit, RIKEN Advanced Science Institute, ²Graduate School of Nanobioscience, Yokohama City University, ³Advanced NMR Metabomics Research Team, RIKEN Plant Science Center, ⁴Graduate School of Bioagricultural Science, Nagoya University

A method for metabolite extraction from microbial planktonic communities is presented. Whole community sampling is achieved by filtration onto specially prepared filters. After lyophilization, aqueous-soluble metabolites are extracted. This approach allows for application of environmental metabolomics to trans-omics investigations of natural or experimental microbial communities.

Herbivore-induced Blueberry Volatiles and Intra-plant Signaling

JoVE 3440 12/18/2011

Cesar R. Rodriguez-Saona

Department of Entomology, Rutgers University

A push-pull method for collecting plant volatiles is described. The method allows for a comparison of volatiles induced by herbivore feeding, exogenous methyl jasmonate, and mechanical damage. This technique is also used to investigate the volatile response of undamaged branches to exposure to volatiles from herbivore-damaged branches within blueberry plants.

Continuously-stirred Anaerobic Digester to Convert Organic Wastes into Biogas: System Setup and Basic Operation

JoVE 3978 7/13/2012

Joseph G. Usack, Catherine M. Spirito, Largus T. Angenent

Department of Biological and Environmental Engineering, Cornell University

Laboratory-scale anaerobic digesters allow scientists to research new ways of optimizing existing applications of anaerobic biotechnology and to evaluate the methane producing potential of various organic wastes. This article introduces a generalized model for the construction, inoculation, operation, and monitoring of a laboratory-scale continuously stirred anaerobic digester.

Investigations on Alterations of Hippocampal Circuit Function Following Mild Traumatic Brain Injury

JoVE 4411 11/19/2012

Colin J. Smith^1,2, Brian N. Johnson¹, Jaclynn A. Elkind¹, Jill M. See¹, Guoxiang Xiong¹, Akiva S. Cohen^1,3

¹Division of Neurology, Children's Hospital of Philadelphia, ²Neuroscience Graduate Group, Perelman School of Medicine at the University of Pennsylvania, ³Department of Pediatrics, Perelman School of Medicine at the University of Pennsylvania

A multi-faceted approach to investigating functional changes to hippocampal circuitry is explained. Electrophysiological techniques are described along with the injury protocol, behavioral testing and regional dissection method. The combination of these techniques can be applied in similar fashion for other brain regions and scientific questions.

Comprehensive Compositional Analysis of Plant Cell Walls (Lignocellulosic biomass) Part II: Carbohydrates

JoVE 1837 3/12/2010

Cliff E. Foster¹, Tina M. Martin¹, Markus Pauly²

¹Great Lakes Bioenergy Research Center, Michigan State University (MSU), ²Great Lakes Bioenergy Research Center and DOE-Plant Research Lab, Michigan State University (MSU)

Plant biomass is a major carbon-neutral renewable resource that could be used for the production of biofuels. Plant biomass consists mainly of cell walls, a structurally complex composite material termed lignocellulosics. Here we describe a protocol for a comprehensive analysis of the content and composition of wall derived carbohydrates.

Arabidopsis thaliana Polar Glycerolipid Profiling by Thin Layer Chromatography (TLC) Coupled with Gas-Liquid Chromatography (GLC)

JoVE 2518 3/18/2011

Zhen Wang, Christoph Benning

Department of Biochemistry and Molecular Biology, Michigan State University

Composition of polar lipid extracts and the fatty acid composition of individual glycerolipids are determined in a simple and robust lipid profiling experiment. For this purpose, glycerolipids are isolated by thin layer chromatography and subjected to transmethylation of their acyl groups. Fatty acyl methylesters are quantified by gas-liquid chromatography.

Assessing Hepatic Metabolic Changes During Progressive Colonization of Germ-free Mouse by ¹H NMR Spectroscopy

JoVE 3642 12/15/2011

Peter Heath¹, Sandrine Paule Claus²

¹School of Chemistry, Food and Pharmacy, The University of Reading, ²Department of Nutritional Sciences, The University of Reading

A progressive colonization procedure is described to further assess its impact on the host hepatic metabolism. Colonization is monitored non invasively by evaluating the urinary excretion of microbial co-metabolites by NMR-based metabolic profiling while hepatic metabolism is assessed by High Resolution Magic Angle Spinning (HR MAS) NMR profiling of intact biopsy.

Large Scale Non-targeted Metabolomic Profiling of Serum by Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS)

JoVE 50242 3/14/2013

Corey D. Broeckling, Adam L. Heuberger, Jessica E. Prenni

Proteomics and Metabolomics Facility, Colorado State University

Non-targeted metabolite profiling by ultra performance liquid chromatography coupled with mass spectrometry (UPLC-MS) is a powerful technique to investigate metabolism. This article outlines a typical workflow utilized for non-targeted metabolite profiling of serum including sample organization and preparation, data acquisition, data analysis, quality control, and metabolite identification.

Compact Quantum Dots for Single-molecule Imaging

JoVE 4236 10/09/2012

Andrew M. Smith¹, Shuming Nie^1,2

¹Department of Biomedical Engineering, Emory University, ²Department of Chemistry, Georgia Institute of Technology

We describe the preparation of colloidal quantum dots with minimized hydrodynamic size for single-molecule fluorescence imaging. Compared to conventional quantum dots, these nanoparticles are similar in size to globular proteins and are optimized for single-molecule brightness, stability against photodegradation, and resistance to nonspecific binding to proteins and cells.

May 2012: This Month in JoVE

JoVE 4464 5/01/2012

Aaron Kolski-Andreaco

Here are some highlights from the May 2012 Issue of Journal of Visualized Experiments (JoVE).

Synthesis and Calibration of Phosphorescent Nanoprobes for Oxygen Imaging in Biological Systems

JoVE 1731 3/03/2010

Louise E. Sinks, Emmanuel Roussakis, Tatiana V. Esipova, Sergei A. Vinogradov

Department of Biochemistry and Biophysics, University of Pennsylvania

We present principles of oxygen measurements by phosphorescence quenching and review design of porphyrin-based dendritic nanosensors for oxygen imaging in biological systems.

Analytical Techniques for Assaying Nitric Oxide Bioactivity

JoVE 3722 6/18/2012

Hong Jiang¹, Deepa Parthasarathy¹, Ashley C. Torregrossa¹, Asad Mian², Nathan S. Bryan¹

¹Texas Therapeutics Institute, University of Texas Health Science Center at Houston, ²Deptartment of Pediatrics, Baylor College of Medicine

The endogenous production of nitric oxide (NO) regulates a wide variety of biological functions. It is becoming increasingly clear that disruption or dysregulation of NO based signaling is involved in many human diseases. Methods to quantify relevant NO metabolites may provide novel diagnostic or prognostic biomarkers for human disease.

Analyzing Large Protein Complexes by Structural Mass Spectrometry

JoVE 1954 6/19/2010

Noam Kirshenbaum, Izhak Michaelevski, Michal Sharon

Department of Biological Chemistry, Weizmann Institute of Science

Mass spectrometry has proven to be a valuable tool for analyzing large protein complexes. This method enables insights into the composition, stoichiometry and overall architecture of multi-subunit assemblies. Here, we describe, step-by-step, how to perform a structural mass spectrometry analysis, and characterize macromolecular structures.

Microwave-assisted Intramolecular Dehydrogenative Diels-Alder Reactions for the Synthesis of Functionalized Naphthalenes/Solvatochromic Dyes

JoVE 50511 4/01/2013

Laura S. Kocsis, Erica Benedetti, Kay M. Brummond

Department of Chemistry, University of Pittsburgh

Microwave-assisted intramolecular dehydrogenative Diels-Alder (DA) reactions provide concise access to functionalized cyclopenta[b]naphthalene building blocks. The utility of this methodology is demonstrated by one-step conversion of the dehydrogenative DA cycloadducts into novel solvatochromic fluorescent dyes via Buchwald-Hartwig palladium-catalyzed cross-coupling reactions.

Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry

JoVE 2812 7/31/2011

Lei Zhao*¹, Jeffrey R. Whiteaker*¹, Matthew E. Pope², Eric Kuhn³, Angela Jackson⁴, N. Leigh Anderson⁵, Terry W. Pearson², Steven A. Carr³, Amanda G. Paulovich¹

¹Clinical Research Division, Fred Hutchinson Cancer Research Center - FHCRC, ²Department of Biochemistry and Microbiology, University of Victoria, ³Broad Institute of MIT and Harvard, ⁴Genome BC Proteomics Centre, University of Victoria, ⁵Plasma Proteome Institute

Stable Isotope Standards and Capture by Anti-Peptide Antibodies (SISCAPA) couples affinity enrichment of peptides with stable isotope dilution mass spectrometry (MRM-MS) to provide quantitative measurement of peptides as surrogates for their respective proteins. Here we describe the protocol using magnetic particles in a partially automated format.

MALDI Imaging Mass Spectrometry of Neuropeptides in Parkinson's Disease

JoVE 3445 2/14/2012

Jörg Hanrieder^1,2, Anna Ljungdahl¹, Malin Andersson¹

¹Department of Pharmaceutical Biosciences, Uppsala University, ²Department of Chemical and Biological Engineering, Chalmers University of Technology

Dopamine replacement pharmacotherapy using L-DOPA is the most commonly used symptomatic treatment of Parkinson’s disease, but is accompanied by side effects including involuntary abnormal movements, termed dyskinesia ¹. Here, a protocol for MALDI imaging mass spectrometry is presented that detects changes in rat brain neuropeptide levels related to dyskinesia.

Production of Disulfide-stabilized Transmembrane Peptide Complexes for Structural Studies

JoVE 50141 3/06/2013

Pooja Sharma^1,2, Mariam Kaywan-Lutfi¹, Logesvaran Krshnan^1,2, Eamon F. X. Byrne^1,2, Melissa Joy Call^1,2, Matthew Edwin Call^1,2

¹Structural Biology Division, The Walter and Eliza Hall Institute of Medical Research, ²The University of Melbourne

Biophysical and biochemical studies of interactions among membrane-embedded protein domains face many technical challenges, the first of which is obtaining appropriate study material. This article describes a protocol for producing and purifying disulfide-stabilized transmembrane peptide complexes that are suitable for structural analysis by solution nuclear magnetic resonance (NMR) and other analytical applications.

Cellular Lipid Extraction for Targeted Stable Isotope Dilution Liquid Chromatography-Mass Spectrometry Analysis

JoVE 3399 11/17/2011

Stacy L. Gelhaus^1,2, A. Clementina Mesaros^1,2, Ian A. Blair^1,2

¹Centers for Cancer Pharmacology and Excellence in Environmental Toxicology, University of Pennsylvania, ²Department of Pharmacology, University of Pennsylvania

This protocol will demonstrate the extraction and analysis of free and esterified bioactive fatty acids from cells. Fatty acids are accurately quantified using stable isotope dilution, chiral liquid chromatography, electron capture atmospheric chemical ionization multiple reaction monitoring mass spectrometry (SID-LC-ECAPCI-MRM/MS).

High-throughput Synthesis of Carbohydrates and Functionalization of Polyanhydride Nanoparticles

JoVE 3967 7/06/2012

Brenda R. Carrillo-Conde*¹, Rajarshi Roychoudhury*², Ana V. Chavez-Santoscoy*¹, Balaji Narasimhan¹, Nicola L.B. Pohl^1,2

¹Department of Chemical and Biological Engineering, Iowa State University, ²Department of Chemistry, Iowa State University

In this article, a high throughput method is presented for the synthesis of oligosaccharides and their attachment to the surface of polyanhydride nanoparticles for further use in targeting specific receptors on antigen presenting cells.

Solid Phase Synthesis of a Functionalized Bis-Peptide Using "Safety Catch" Methodology

JoVE 4112 5/15/2012

Conrad T. Pfeiffer, Christian E. Schafmeister

College of Science and Technology, Temple University

The efficient solid-phase peptide synthesis of a functionalized bis-peptide trimer utilizing a "safety catch" cleavage procedure from HMBA resin is described.

Monitoring Protein Adsorption with Solid-state Nanopores

JoVE 3560 12/02/2011

David J. Niedzwiecki, Liviu Movileanu

Department of Physics, Syracuse University

A method of using solid-state nanopores to monitor the non-specific adsorption of proteins onto an inorganic surface is described. The method employs the resistive-pulse principle, allowing for the adsorption to be probed in real-time and at the single-molecule level. Because the process of single protein adsorption is far from equilibrium, we propose the employment of parallel arrays of synthetic nanopores, enabling for the quantitative determination of the apparent first-order reaction rate constant of protein adsorption as well as and the Langmuir adsorption constant.

Examining BCL-2 Family Function with Large Unilamellar Vesicles

JoVE 4291 10/05/2012

James J. Asciolla, Thibaud T. Renault, Jerry E. Chipuk

Department of Oncological Sciences, Department of Dermatology, The Tisch Cancer Institute, The Graduate School of Biological Sciences, Mount Sinai School of Medicine

Biochemically-defined large unilamellar vesicles (LUVs) are a convenient model system to analyze BCL-2 family interactions with immediate implications in better understanding the mitochondrial pathway of apoptosis. A method to produce LUVs, along with standard BCL-2 family protein combinations and controls to examine LUV permeabilization, are presented.

Viral Nanoparticles for In vivo Tumor Imaging

JoVE 4352 11/16/2012

Amy M. Wen¹, Karin L. Lee¹, Ibrahim Yildiz¹, Michael A. Bruckman¹, Sourabh Shukla¹, Nicole F. Steinmetz²

¹Department of Biomedical Engineering, Case Western Reserve University, ²Department of Biomedical Engineering, Radiology, and Materials Science and Engineering, Case Western Reserve University

Plant viral nanoparticles (VNPs) are promising platforms for applications in biomedicine. Here, we describe the procedures for plant VNP propagation, purification, characterization, and bioconjugation. Finally, we show the application of VNPs for tumor homing and imaging using a mouse xenograft model and fluorescence imaging.

Methods Development for Blood Borne Macrophage Carriage of Nanoformulated Antiretroviral Drugs

JoVE 2460 12/09/2010

Shantanu Balkundi*, Ari S. Nowacek*, Upal Roy, Andrea Martinez-Skinner, JoEllyn McMillan, Howard E. Gendelman

Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center

Nanoparticles of indinavir, ritonavir, efavirenz and atazanavir were manufactured using wet milling, homogenization and ultrasonication. These nanoformulations, collectively termed nanoformulated antiretroviral therapy (nanoART), assessed macrophage-based drug delivery. Monocyte-derived macrophage nanoART uptake, retention and sustained release were determined. These preliminary studies suggest the potential of nanoART for clinical use.