Design and evaluation of a CXCR4 targeting peptide 4DV3 as an HIV entry inhibitor and a ligand for targeted drug delivery.

The feasibility of utilizing the cell surface chemokine receptor CXCR4 for human immunodeficiency virus (HIV) entry inhibition and as an intracellular portal for targeted drug delivery was evaluated. Novel DV3 ligands (1DV3, 2DV3, and 4DV3) were designed, synthesized and conjugated to various probes (fluorescein isothiocyanate (FITC) or biotin) and cargos with sizes ranging from 10 to 50 nm (polyethylene glycol (PEG), streptavidin, and a polymeric nanoparticle). 4DV3 conjugated probes inhibited HIV-1 entry into the CXCR4-expressing reporter cell line TZM-bl (IC50 at 553 nM) whereas 1DV3 and 2DV3 did not. 4DV3 also inhibited binding of anti-CXCR4 antibody 44,708 to TZM-bl cells with nanomolar potency, while the small-molecule CXCR4 antagonist AMD3100 did not. Molecular modeling suggested simultaneous binding of a single 4DV3 molecule to four CXCR4 molecules. Differences in CXCR4-binding sites could explain the discrete inhibitory effects observed for 4DV3, the 44,708 antibody and AMD3100. In the Sup-T1 cell chemotaxis assay, the 4DV3 ligand functioned as a CXCR4 allosteric enhancer. In addition, 4DV3 ligand-conjugated cargos with sizes ranging from 10 to 50 nm were taken up into CXCR4-expressing Sup-T1 and TZM-bl cells, demonstrating that CXCR4 could serve as a drug delivery portal for nanocarriers. The uptake of 4DV3 functionalized nanocarriers combined with the allosteric interaction with CXCR4 suggests enhanced endocytosis occurs when 4DV3 is the targeting ligand. The current results indicate that 4DV3 might serve as a prototype for a new type of dual function ligand, one that acts as a HIV-1 entry inhibitor and a CXCR4 drug delivery targeting ligand.

Investigation on Sized-Regulated Iron Nanoparticles Prepared by Liquid Phase Plasma Reduction Process.

The liquid-phase plasma reduction method has been applied to prepare iron nanoparticles from iron chloride solution using a bipolar pulsed electrical discharge system. The excited states of atomic iron, hydrogen, and oxygen as well as the molecular bands of hydroxyl radicals were detected in the emission spectra. The iron nanoclusters formed at the initial stage convert to dispersion of small iron nanoparticles, which then grows slowly to form anisotropic, tetragonal shape. The cationic surfactant of CTAB was shown to exhibit a large influence on the particle generation procedure.

Diffusion tensor microscopy in human nervous tissue with quantitative correlation based on direct histological comparison.

Thanks to its proven utility in both clinical and research applications, diffusion tensor tractography (DTT) is regularly employed as a means of delineating white-matter tracts. While successful efforts have been made to validate tractographic predictions, comparative methods which would permit the validation of such predictions at microscopic resolutions in complex biological tissues have remained elusive. In a previous study, we attempted to validate for the first time such predictions at microscopic resolutions in rat and pig spinal cords using a semi-quantitative analysis method. In the current study, we report improved quantitative analysis methods that can be used to determine the accuracy of DTT through comparative histology and apply these techniques for the first time to human tissue (spinal cord) samples. Histological images are down-sampled to resolutions equivalent to our magnetic resonance microscopy (MRM) and converted to binary maps using an automated thresholding tool. These maps (n=3) are co-registered to the MRM allowing us to quantify the agreement based on the number of pixels which contain tracts common to both imaging datasets. In our experiments, we find that-on average-89% of imaging pixels predicted by DTT to contain in-plane white-matter tract structure correspond to physical tracts identified by histology. In addition, angular analysis comparing the orientation of fiber tracts measured in histology to their corresponding in-plane primary eigenvector components is presented. Thus, as well as demonstrating feasibility in human tissue, we report a robust agreement between imaging datasets taken at microscopic resolution and confirm the primary eigenvector's role as a fundamental parameter with clear physical correlates in the microscopic regime.

Application of phage display to discovery of tumor-specific homing peptides: developing strategies for therapy and molecular imaging of cancer.

Selective delivery of drugs or imaging dyes to tumor is the central challenge for improving existing therapy and diagnosis of cancer. Phage display of random peptides has been used to identify homing peptides that are specific to tumor cells or tumor blood vessels. These homing peptides can be conjugated with imaging agents or therapeutic drugs and may be a promising tool for affinity-based targeted delivery of imaging agents and drugs. This chapter describes the procedures for phage display selection of peptides that selectively bind to bladder tumor cells and home to tumor tissues in vivo. It also describes the application of the tumor-specific peptide to the detection of bladder cancer cells in the patient urine.

Highly sensitive "turn-on" fluorescent sensor for Hg2+ in aqueous solution based on structure-switching DNA.

A simple design of "turn-on" fluorescent sensor for mercury was demonstrated based on structure-switching DNA with a low detection limit of 3.2 nM and high selectivity.

Cloning and characterization of FAM13A1--a gene near a milk protein QTL on BTA6: evidence for population-wide linkage disequilibrium in Israeli Holsteins.

A cluster of genes coding for proteins of the extracellular matrix (ECM) containing sequence motifs essential for integrin-receptor interactions is located on HSA4q21 and on BTA6, within the critical region of a quantitative trait locus (QTL) affecting milk protein production. Genes within this cluster are involved in the formation of bone and lobuloalveolar structures in mammary gland and in kidney function. We cloned a bovine gene neighboring this ECM cluster, termed FAM13A1, the first member of a novel gene family (FAM13). A short predominant 5.1-kb mRNA variant capable of encoding 697 amino acids is transcribed from 18-exon orthologous genes in human and mouse. All putative protein orthologs contained a bipartite nuclear-localization signal and two coil-coiled domains. We detected two other FAM13 paralogs, C5ORF5 and FAM13C1, on HSA5q31 and HSA10q21, respectively. All FAM13 paralogs produce transcripts that are complementary to adjacent genes, suggesting that antisense transcription may regulate their functions. The structure of a longer 5.9-kb variant, FAM13A1_v2, that has an extra 7-exon putative RhoGAP domain at the N-terminus provides further clues to FAM13 function. Analysis of 400 bulls revealed a population-wide linkage disequilibrium between FAM13A1 polymorphisms and the QTL.

New sensitive determination method of benzidine-hemoglobin adducts by gas chromatography-electron impact mass spectrometry.

A gas chromatographic-mass spectrometric assay was developed for the determination of benzidine (BZ)-hemoglobin adducts. Adducts were released from hemoglobin by alkaline hydrolysis and extraction at pH 8 with ethyl ether. The dried extract was completely derivatized with N-methyl-N-(tert.-butyldimethylsilyl) trifluoroacetamide (MTBDMSTFA)-NH(4)I (1000:3) under catalysis of dithioerythritol. The recovery of BZ, acetylbenzidine (ABZ) and diacetylbenzidine (DABZ) in the extraction procedure was 76-98%. The detection limits of the assay were 0.1 ng/g for both BZ and ABZ, and 0.5 ng/g for DABZ based upon assayed hemoglobin of 0.1 g. The method was applied to the determination of BZ-hemoglobin adducts formed in young female Sprague-Dawley rats after treatment for 1, 2 and 3 weeks with 0.008% BZ via the drinking water. Two adducts were detected by proposed procedure. The structure of these adducts could be assigned to BZ and ABZ. After 1 week, the total mean amount of adducts determined was 2.8 ng/g hemoglobin. The adduct levels increased up to about 7.5 ng/g after a week and, thereafter, remained essentially constant. The relative contribution of BZ and ABZ to the total hemoglobin adduct level was strongly treatment time-dependent. After 1 week, the BZ and ABZ adducts were formed at similar levels, whereas after 3 weeks the ABZ adducts was predominant. Treatment of rats for 3 weeks in the dose range 12.2-36.8 mg of BZ in drinking water resulted in a dose-proportional increase in the total amount of hemoglobin adducts formed.

Patient satisfaction with telemedicine in home health services for the elderly.

In a pilot study of telemedicine in home health services (HHS) for elderly patients, we implemented and evaluated a telemedicine system with a 33-kbs narrow-band approach to determine its effectiveness in providing quality services. Fifty patients were selected for the study. We found that telemedicine was effective in terms of reducing the number of clinic visits and achieving patient satisfaction. The average number of clinic visits per month was significantly decreased from 0.64 to 0.42 (p < 0.05) after the use of telemedicine. 72% of patients were satisfied with telemedicine, but only patient location showed a significant difference for patient satisfaction (p < 0.05). Patients in their homes (82%) were more satisfied than patients in nursing homes (50%). Of four types of services provided, medical consultation (100%) was the most highly satisfactory service with telemedicine, followed by physical therapy (83.3%). Although the satisfaction scores did not indicate a significant difference in the system characteristics, the quality of verbal communication appeared to be a more important factor in influencing patient satisfaction than set-up time or quality of image. A computer-based patient record was also developed to view a patient summary and to document encounters at the patient's home. Since the system is a low-cost approach that is easy to interface with a notebook computer, it can support various other HHSs.