H double-quantum filtered MR imaging of joints tissues: bound water specific imaging of tendons, ligaments and cartilage

The ¹H double-quantum filtered (DQF) NMR and DQF MRI is applied to the joint tissues of rabbits for selective visualization of tendons, menisci and articular cartilage. The ¹H DQF NMR selectively filters double-quantum coherence arising from the ¹H dipolar interaction of the “bound” water in these tissues. The double-quantum creation time dependency of the DQF signal intensity is determined by the molecular environment of the “bound” water. Therefore, each tissue has a unique creation time at which the DQF signal reaches its maximum intensity, τ_max (Achilles tendon: 0.46 ± 0.02 ms, patella: 0.55 ± 0.8 ms, anterior cruciate ligament: 0.60 ± 0.05 ms, meniscus: 0.78 ± 0.02 ms, skin: 0.81 ± 0.07 ms). We have presented the creation-time-contrasted DQF images of the meniscus, patella, foot, and knee joint. Compared with conventional T₂*-weighted gradient-echo (GRE) MR images, tendons, ligaments, menisci, and articular cartilage were more clearly seen in the DQF MR images. All these tissues were distinctly discriminated from each other by their creation times. DQF MR images of foot and knee joints can selectively demonstrated tendons, ligaments, and cartilage, which make it easier to understand the complicated anatomic structure of joints. Because the DQF NMR signal intensity and τ_max are sensitive to the order structure of the “bound” water, it might be possible to introduce the creation-time dependent-contrast of ¹H DQF MR images as a new tool for analyzing the changes in the ordered structure of the tissue.     

Synthesis and Cytotoxic Activity of Macromolecular Prodrug of Cisplatin Using Poly(ethylene glycol) with Galactose Residues or Antennary Galactose Units

To provide a macromolecular prodrug with recognition ability for hepatoma cells, we synthesized new conjugates of cisplatin (CDDP) and poly(ethylene glycol) (PEG) with galactose residues or antennary galactose units (Gal4A, four branched galactose residues) at the chain terminus, Gal‐PEG‐DA/CDDP or Gal4A‐PEG‐DA/CDDP conjugates. An antennary (branched) structure of Gal4A was designed based on the fact that saccharide clusters with branched structures show highly effective binding with saccharide receptors, a phenomenon known as the ‘cluster effect’. The cytotoxic activity of the conjugates was investigated against HepG2 human hepatoma cells in vitro and compared with a control conjugate without galactose, MeO‐PEG‐DA/CDDP. Gal‐PEG‐DA/CDDP and Gal4A‐PEG‐DA/CDDP conjugates showed lower IC₅₀ values (3.1×10^–4 and 2.3×10^–4 M , respectively) than the MeO‐PEG‐DA/CDDP conjugate (10.5×10^–4 M ). The cytotoxic activities of these conjugates with galactose residues or antennary galactose units were inhibited as a result of the addition of galactose and strongly inhibited by the addition of Gal4A, however the inclusion of a methoxy group (the MeO‐PEG‐DA/CDDP conjugate) did not affect the activity. These results suggest that the Gal4A unit introduced to the conjugate has effective recognition ability against HepG2 human hepatoma cells.     

Synthesis and functionalities of poly(N‐vinylalkylamide). XII. Synthesis and thermosensitive property of poly(vinylamine) copolymer prepared from poly(N‐vinylformamide‐co‐N‐vinylisobutyramide)

The differences in the polymerization abilities of N‐vinylformamide (NVF) and N‐vinylisobutyramide (NVIBA) and the synthesis of their copolymers were studied. The polymerization abilities were fairly good and quite similar to those of N‐vinyl‐ acetamide (NVA), a monomer in the same class as N‐vinylalkylamides. Since the monomer reactivity ratios were r₁ = 1.08 and r₂ = 0.92 (M₁ = NVF, M₂ = NVIBA), respectively, it is clear that the comonomers definitely were converted to random copolymers. The resulting copolymers poly(NVF‐co‐NVIBA) exhibited the cloud points sharply. The light transmittance profiles were the same as those for poly(NVIBA) although they increased from 39 °C for poly(NVIBA), with an increase in the corresponding hydrophilic NVF component. Our final objective was to produce a cloud point controlled polymer material with primary amino groups. To achieve this, we examined the hydrolysis of poly(NVF), poly(NVA), poly(NVIBA), and poly(NVF‐co‐NVIBA) to obtain poly(vinylamine) [poly(VAm)]. The hydrolytic cleavage of poly(NVF) and poly(NVA) was promoted by an increase in temperature. However, poly(NVIBA) was not cleaved appreciably. The hydrolysis of poly(NVF‐co‐NVIBA) was done under controlled conditions, and amino groups selectively were introduced to only one of two components of the copolymer. The cloud point of the hydrolyzed copolymer shifted to a higher temperature than that of the copolymer. © 2000 John Wiley & Sons, Inc. J Polym Sci A: Polym Chem 38: 3674–3681, 2000     

Metal‐imprinted microsphere prepared by surface template polymerization and its application to chromatography

A metal ion‐imprinted microsphere was prepared by surface molecular template polymerization. Trimethylolpropane trimethacrylate (TRIM), zinc ions, 1,12‐dodecanediol‐O, O′‐diphenyl phosphonic acid (DDDPA) were used as a crosslinking agent, an imprint molecule, and a functional host molecule. The Zn(II)‐imprinted microspheres, which are spherically well‐defined particles, were prepared by using water‐in‐oil‐in‐water (W/O/W) multiple emulsions. The combination of TRIM and DDDPA serves to align the recognition sites resulting in better template sites produced on the polymer surface. We firstly conducted diagnostic zinc‐ and copper‐ion adsorption tests with the Zn(II)‐imprinted and unimprinted microspheres in order to make an assessment on the effectiveness of the molecular imprinting technique. Further, the metal‐imprinted microspheres were applied to the column operation. The separation and recovery of metals were carried out by an adsorption column packed with the Zn(II)‐imprinted microspheres. This performance was compared to that of commercial chelating resins that possess similar phosphoric functional groups. The Zn(II)‐imprinted polymer shows an extremely high selectivity to the imprinted zinc ions compared to that of the commercial chelating resin. © 2000 John Wiley & Sons, Inc. J Polym Sci A: Polym Chem 38: 689–696, 2000     

Development of a Versatile Protein Labeling Tool for Live-Cell Imaging Using Fluorescent β-Lactamase Inhibitors

To understand the function of protein in live cells, real-time monitoring of protein dynamics and sensing of their surrounding environment are important methods. Fluorescent labeling tools are thus needed that possess fast labeling kinetics, high efficiency, and long-term stability. We developed a versatile chemical protein-labeling tool based on fluorophore-conjugated diazabicyclooctane β-lactamase inhibitors (BLIs) and wild-type TEM-1 β-lactamase protein tag. The fluorescent probes efficiently formed a stable carbamoylated complex with β-lactamase, and the labeled proteins were visualized over a long period of time in live cells. Moreover, use of an α-fluorinated carboxylate ester-based BLI prodrug enabled the probe to permeate cell membranes and stably label intracellular proteins after unexpected spontaneous ester hydrolysis. Lastly, combining the labeling tool with a pH-activatable fluorescent probe allowed visual monitoring of lysosomal protein translocation during autophagy.     

Recent Advances in Hybrid Materials of Metal Nanoparticles and Polyoxometalates

Polyoxometalates (POMs), anionic metal-oxygen nanoclusters that possess various composition-dependent properties, are widely used to modify the existing properties of metal nanoparticles and to endow them with new ones. Herein, we present an overview of recent advances in hybrid materials that consist of metal nanoparticles and POMs. Following a brief introduction on the inception of this area and its development, representative properties and applications of these materials in various fields such as electrochemistry, photochemistry, and catalysis are introduced. We discuss how the combination of two classic inorganic materials facilitates cooperative and synergistic behavior, and we also give personal perspectives on the future development of this field.     

Improvement of Antibody Activity by Controlling Its Dynamics Using the Glycan–Lectin Interaction

Antibody dynamics on membranes, such as endocytosis and clustering, are vital in determining antibody functions. In this study, we demonstrated that glycan conjugation can modulate antibody dynamics through the glycan–lectin interaction to regulate its potency. The anti-HER2 antibody, an anti-breast-cancer antibody, was conjugated with galactose-containing N-glycan, and its internalization was suppressed by interaction with galectin-3, leading to enhanced complement-dependent cytotoxic (CDC) activity. This glycan–antibody conjugate is proposed as a new approach to modulate antibody activity and may provide an alternative strategy for redeveloping antibody drugs that do not exhibit sufficient activity.     

Evaluation of the accuracy of hTERT gene aberrant methylation using electrochemical hybridization assay and liquid-based cytology in screening for oral squamous cell carcinoma

In order to improve the survival rate of oral squamous cell carcinoma (OSCC) patients, a reliable diagnostic method for early OSCC detection is required that is minimally invasive, less burdensome to the patient, and has high sensitivity and specificity. Therefore, we performed the detection of abnormal methylation at three locations in the hTERT promoter region of oral exfoliated cells by employing the ferrocenylnaphthalene diimide (FND)-based electrochemical hybridization assay (FND-EHA) using three types of DNA probe-immobilized electrodes. We also performed liquid cytology using oral exfoliated cells and compared these obtained data to evaluate whether FND- EHA can be used as an OSCC screening system. The results showed a good correlation between this method and conventional OSCC screening, and cytology. In addition, FND-EHA was also able to determine samples that had been ambiguously determined by liquid cytology. This indicates that FND-EHA may be useful as an OSCC screening system.