Highly selective fluorometric determination of polyamines based on intramolecular excimer-forming derivatization with a pyrene-labeling reagent

We introduce a novel approach in highly selective and sensitive fluorescence derivatization of polyamines. This method is based on an intramolecular excimer-forming fluorescence derivatization with a pyrene reagent, 4-(1-pyrene)butyric acid N-hydroxysuccinimide ester (PSE), followed by reversed-phase high-performance liquid chromatography (HPLC). Polyamines, having two to four amino moieties in a molecule, were converted to the corresponding dipyrene- to tetrapyrene-labeled derivatives by reaction (100 degrees C, 20 min) with PSE. The derivatives afforded intramolecular excimer fluorescence (450-520 nm), which can clearly be discriminated from the monomer (normal) fluorescence (360-420 nm) emitted from PSE, its hydrolysate and monopyrene-labeled derivatives of monoamines. The structures of the derivatives were confirmed by HPLC with mass spectrometry, and the emission of excimer fluorescence could be proved by spectrofluorometry and time-resolved fluorometry. The PSE derivatives of four polyamines [putrescine (Put), cadaverine (Cad), spermidine (Spd), and spermine (Spm)] could be separated by reversed-phase HPLC on a C8 column with linear gradient elution. The detection limits (signal-to-noise ratio of 3) for the polyamines were 1 (Put), 1 (Cad), 5 (Spd), and 8 (Spm) fmol on the column. Furthermore, the present method was so selective that biogenic monoamines gave no peak in the chromatogram.     

Characterization of natural resin shellac by reactive pyrolysis-gas chromatography in the presence of organic alkali

A method to determine chemical composition of natural resin shellac was developed on the basis of reactive pyrolysis-gas chromatography (Py-GC) in the presence of an organic alkali, tetramethylammonium hydroxide ((CH(3))(4)NOH, TMAH). Py-GC using 25% TMAH aqueous solution enabled the highly sensitive determination of terpenic acids, aleuritic acid, several minor fatty acids, and the wax components of shellac as their methyl derivatives on the resulting pyrograms with less than 2.0% relative standard deviations without using any cumbersome pretreatment. The observed average distributions of each resin acid component determined by reactive Py-GC for shellac samples from India and Thailand showed that the average ratios among terpenic acids, aleuritic acid, and the other fatty acids were about 53:34:14 for Indian shellac and 51:35:14 for Thailand shellac, respectively, suggesting a slightly significant difference. However, clearer discrimination of the shellac samples from the two different growing places was attainable by applying principal component analysis for the mole percent distributions of all the acidic components determined by reactive Py-GC.     

17O Knight shift study in the superconducting state of Sr2RuO4

¹⁷O Knight shift measurements in Sr₂RuO₄ were performed over the wide range of magnetic field 3.2-11.4kOe parallel to the basal RuO₂ planes. The spin susceptibility is totally unchanged through its T_c, evidencing that the spin-triplet superconducting state is realized in Sr₂RuO₄. The result indicates that the Cooper pairs consist of the parallel spin pairs | > and | > with their quantization axis perpendicular to the c-axis direction. The in-plane 2D nearly ferromagnetic spin fluctuations may play a role for the stabilization of this state among various representations of spin-triplet order parameter.     

Fas-independent and nonapoptotic cytotoxicity mediated by a human CD4(+) T-cell clone directed against an acute myelogenous leukemia-associated DEK-CAN fusion peptide

The mechanism underlying the cytotoxicity mediated by a human CD4(+) cytotoxic T-lymphocyte (CTL) clone directed against a peptide derived from the acute myelogenous leukemia-associated fusion protein, DEK-CAN, was investigated. A DEK-CAN fusion peptide-specific CD4(+) Th0 CTL clone, designated HO-1, was established from the peripheral blood lymphocytes of a healthy individual. HO-1 exerted direct but not "innocent bystander" cytotoxicity within 2 hours. The cytotoxicity mediated by HO-1 was completely Ca2+-dependent. Because HO-1 lysed peptide-loaded Fas-deficient target cells derived from a patient with a homozygous Fas gene mutation, its cytotoxicity appeared to be mediated by a Fas-independent pathway. In addition, its cytotoxicity was only partially inhibited by treatment with concanamycin A and strontium ions, which are inhibitors of the perforin-based cytotoxic pathway. Although membrane-bound type of tumor necrosis factor-alpha (TNF-alpha) was expressed on HO-1, an anti-TNF-alpha antibody had no effect on HO-1-mediated cytotoxicity. HO-1 expressed mRNA for apoptosis-inducing mediators, including perforin, granzyme B, Fas ligand, TNF-alpha, and lymphotoxin; however, no DNA fragmentation was detected in target cells incubated with HO-1 by 5-[125I]Iodo-2'-deoxyuridine release assay and agarose gel electrophoresis of DNA. Although it has been suggested that the Fas/Fas ligand system is the main pathway by which CD4(+) CTL-mediated cytotoxicity is exerted in murine systems, HO-1 produced peptide-specific and HLA-restricted cytotoxicity via a Fas-independent and nonapoptotic pathway. The present study thus describes a novel mechanism of cytotoxicity mediated by CD4(+) CTL.     

Regulation of selectin binding activity by cyclization of sialic acid moiety of carbohydrate ligands on human leukocytes

We provide here evidence that supports the occurrence of a biologically dormant form of selectin ligand carbohydrate, the sialyl 6-sulfo Lewis X containing modified sialic acid, in human leukocytes. The modification of sialic acid involves first de-N-acetylation of sialic acid moiety through ubiquitous de-N-acetylation/re-N-acetylation cycle, followed by the dehydrative cyclization of de-N-acetyl sialic acid to form "cyclic sialic acid." The enzyme involved in the dehydration of de-N-acetyl sialic acid is a calcium-dependent enzyme having neutral-alkaline pH optimum. De-N-acetyl sialyl 6-sulfo Lewis X retained selectin binding activity as well as parental sialyl 6-sulfo Lewis X, but cyclic sialyl 6-sulfo Lewis X was devoid of selectin binding activity. Sialyl 6-sulfo Lewis X carrying the cyclic sialic acid is specifically recognized by the newly generated mAb, G159. The determinant was distributed widely among normal human leukocytes, especially on monocytes and subsets of lymphocytes including NK cells, helper memory T cells, Tcr-gammadelta T cells, and a part of B cells. The determinant was detected also on several cultured lymphocytic leukemia cell lines and O-tetradecanoylphorbol 13-acetate-activated lymphoid cells. Cyclic sialyl 6-sulfo Lewis X is efficiently formed by the action of the partly membrane-bound calcium-dependent enzyme, tentatively called "sialic acid cyclase," and a possible physiological significance of this reaction could be a rapid inactivation of selectin binding activity at the cell surface. Conversely, the accumulated intracellular cyclic sialyl 6-sulfo Lewis X determinant may function as a dormant pool of selectin ligands, which, on appropriate stimulation, is hydrolyzed and becomes active in selectin-dependent cell adhesion.     

Effects of age and sex on sodium chloride sensitivity: association with plasma renin activity

The mechanism by which excessive NaCl intake raises blood pressure has not been fully clarified. The present study was undertaken in 87 Japanese inpatients with essential hypertension to investigate the interrelation among effects of age, sex and the renin-angiotensin system on NaCl sensitivity. After ingesting a regular NaCl diet (170 mmol/day) for one week, subjects were placed sequentially on a week of low NaCl diet (50 mmol/day) and a week of high NaCl diet (340 mmol/day). NaCl sensitivity defined as the difference in mean blood pressure between the low and high NaCl diets did not differ between genders. NaCl sensitivity was positively correlated with age and the change in PRA. The fall in PRA after NaCl loading was significantly smaller in women than in men. By multiple regression analysis, age and the change in PRA independently contributed to the change in mean blood pressure. Furthermore, the interaction between sex and the change in PRA was selected as a statistically significant variable. In conclusion, NaCl sensitivity of blood pressure is independently associated with age and the inadequate suppression of the renin-angiotensin system. Because the contribution of the change in PRA to NaCl sensitivity was greater in women than in men, the mechanism of blood pressure elevation after NaCl loading may differ between genders.     

Real-time output voltage control method of quasi-ZCS seriesresonant HF-linked DC-AC converter

This paper proposes a real-time control method of a series resonant high-frequency linked DC-AC converter employing quasi-zero current switching (quasi-ZCS) and a feedback control method for sinusoidal output voltage. An approximate analysis of the converter is performed, and then simplified equations and an equivalent circuit similar to the conventional PWM inverters/inverters are obtained. A real-time feedback control of the converter is realized using the equivalent circuit without detecting HF link current. The usefulness of the proposed control algorithm is confirmed by experimental results     

GaAs field-effect transistors by selective sulphur-ion implantation

GaAs field-effect transistors without a mesa structure have been fabricated by selective sulphur-ion implantation into Cr-doped GaAs substrates. The transconductance was 16 mS and the maximum oscillation frequency was 30 GHz.     

Formation of silica and silicates during the solidification of Fe−Si−O alloys

Small (35 g) ingots of Fe?Si?O alloys have been solidified under controlled conditions and the form, distribution, and compositions of the oxides in the cast structures are reported. The composition of a melt was established by charging a preselected amount of silicon, 0.002 < pct Si < 0.7, and equilibrating the melt with the silica crucible at 1550° or 1650°C. The concentration of FeO and the morphology of the oxide particles in the cast structures indicate that during solidification the process of segregation causes the oxygen potential in the interdendritic liquid to increase over that in equilibrium with the silicon content of the liquid. It appears that nuclei for the formation of the oxide phases, which probably are in the liquid prior to the onset of solidification, become surrounded by solid iron and are made ineffective. It is possible at lower silicon levels that homogeneous nucleation may result by the melt becoming sufficiently supersaturated in oxygen to form FeO or an iron-silicate liquid phase.