Urinary albumin and transferrin as early diagnostic markers of chronic kidney disease.

Feline renal diseases are increasingly noted in veterinary practice. It is important to diagnose and identify the pathological basis of renal dysfunction accurately at an early stage, but there are only a few reports on this area in clinical veterinary medicine. We investigated the efficacy of measurement of urinary albumin (u-Alb) and urinary transferrin (u-Tf) for early diagnosis using 5-µl urine samples collected noninvasively by catheterization from normal (IRIS stage I) cats and cats with stage I chronic kidney disease (CKD). The u-Alb levels in normal and stage I CKD cats were 6.0 ± 4.5 and 11.2 ± 8.4 mg/dl, respectively, and the u-Tf levels were 0.09 ± 0.42 and 0.52 ± 0.79 mg/dl, respectively. Based on ROC curve analysis, the sensitivity and specificity of u-Alb and u-Tf were higher than those of the currently used biomarker, the plasma creatinine level. The sensitivity of u-Alb was higher than that of u-Tf, whereas the specificity of u-Tf was higher than that of u-Alb. The validity of the threshold albumin level (20 mg/dl) was confirmed by measurements using SDS-PAGE. Since leakage of u-Tf in urine precedes leakage of u-Alb, inclusion of u-Tf in biochemistry tests may be appropriate for IRIS staging as a diagnostic marker of early diagnosis of renal disorder in cats.

Effective methods for the measurement of CsI cluster ions using MALDI-MS with suitable solvent combinations and additives.

A method to measure CsI cluster ions ((CsI)(n)Cs(+), (CsI)(n)I(-)) from CsI samples in matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) was developed with a 2-[(2E)-3-(4-tert-butylphenyl)-2-methylprop-2-enylidene] malononitrile (DCTB) matrix and additives. Solvent combinations in which the CsI and DCTB solutions were miscible were effective in detecting CsI cluster ions at a mass range of over m/z 2000 and are associated with a characteristic spread of DCTB within the CsI/DCTB mixture. The addition of saccharides or sugar alcohols to the CsI/DCTB mixture improved the DCTB distribution and widened the mass distribution of CsI cluster ions up to m/z 10,000 in the linear mode.

WITHDRAWN: Solvent effect on PCR from a viewpoint of a change of microscopic environment of Mg(2+) in a solution.

This article has been withdrawn at the request of the editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.

Nature of the chemical bond formed with the structural metal ion at the A9/G10.1 motif derived from hammerhead ribozymes.

We have studied the interaction between metal ions and the metal ion-binding motif in hammerhead ribozymes, as well as the functions of the metal ion at the motif, with heteronuclear NMR spectroscopy. In this study, we employed model RNA systems which mimic the metal ion-binding motif and the altered motif. In Co(NH3)6(III) titrations, we observed large 1H and 31P chemical shift perturbations for the motif and found that outer-sphere complexation of Co(NH3)6(III) is possible for this motif. From the reinvestigation of our previous 15N chemical shift data for Cd(II) binding, in comparison with those of organometallic compounds, we conclude that Cd(II) can form an inner-sphere complex with the nucleobase in the motif. Therefore, the A9/G10.1 site was found to accept both inner-sphere and outer-sphere complexations. The Mg(II) titration for a slightly different motif from the A9/G10.1 site (G10.1-C11.1 to A10.1-U11.1) revealed that its affinity to Mg(II) was drastically reduced, although the ribozyme with this altered motif is known to retain enzymatic activities. This observation suggests that the metal ion at these motifs is not a catalytic center of hammerhead ribozymes.

Microscopic environment of metal ion controlled by the balance between preferential solvation and coordination.

The preferential solvation and the coordination characterizing metal ions (Mg2+ and Zn2+) in solution, which control the microscopic environments around the metal ions, were directly observed through the mass spectrometric analysis of clusters isolated from liquid droplets.