A peroxyoxalate chemiluminescence detection in high-performance liquid chromatography was examined for determination of long chain aliphatic aldehydes using 4-(N,N-dimethylaminosulphonyl)-7-hydrazino-2,1,3-benzoxadiazole (DBD-H) as a fluorogenic labelling reagent. A mixture of 0.6 mM bis(2,4,6-trichlorophenyl)oxalate and 100 mM H2O2 in acetonitrile was used as a post-column chemiluminescent reagent. DBD-hydrazones of five long chain aliphatic aldehydes (C11-C18:1) were eluted and well separated within 16 min with a mobile phase of 5 mM imidazole in acetonitrile. The detection limits were 14-18 fmol at a signal-to-noise ratio (S/N) of 2. DBD-hydrazones of four aliphatic aldehydes (C6-C11) were also eluted and well separated within 30 min with a mobile phase of 5mM imidazole buffer (pH 7.5)-acetonitrile (20/80, v/v). The detection limits were 86-152 fmol (S/N = 2).

Aldehydes↗

PubMedDate not supplied

Flow-injection analysis of cobalt(II) utilizing enhanced lophine chemiluminescence with hydroxylammonium chloride.

A rapid and convenient flow-injection method is described for the determination of Co(II). The method utilized the phenomenon that the lophine-Co(II)-H2O2 chemiluminescence (CL) reaction is enhanced in alkaline media by the addition of hydroxylammonium chloride. The calibration curve was linear over the range 2 x 10(-7) to 2 x 10(-5) M with a detection limit of 4.5 x 10(-8) M (0.9 pmol) at a signal-to-noise ratio of 2. The method was applied to the assay of cyanocobalamin in commercially available eye lotions.

Chromatography, Liquid↗

PubMedDate not supplied

Chemiluminescent derivatization of adenyl compounds with glyoxal derivatives in the presence of heteropoly acids and its application to the simple and sensitive determination of DNA.

A chemiluminescence (CL) determination of adenyl compounds is described. CL derivatization of adenyl compounds with methylglyoxal dimethyl acetal was performed in the presence of tungstosilicic acid and propan-2-ol. CL from adenyl compounds was produced by hydrogen peroxide and L-cysteine ethyl ester in DMF and water. The proposed method is highly sensitive and specific to compounds containing adenine. Adenine was determined in the range 1.0 x 10(-3)-5.0 x 10(-8) M with the detection limit of 3.0 x 10(-8) M (150 fmol per assay). The method was applied to the determination of DNA and detection limits of a few nanograms of DNA achieved.

Acetone↗

PubMedDate not supplied

Photographic detection of fluorescent-labelled oligodeoxynucleotide in the blotting format by peroxyoxalate chemiluminescence.

The preparation of a fluorescent labelled oligonucleotide and its photographic detection by peroxyoxalate chemiluminescence (PO-CL) are described. Fluorescent labelling of an oligonucleotide (15-mer) was performed with naphthalene-2,3-dicarboxaldehyde to give an N-substituted 1-cyanobenz[f]isoindole (CBI) derivative (CBI-15-mer). For the photographic detection of CBI-15-mer, the bis(2,6-difluorophenyl) oxalate (DFPO)-dimethyl phthalate (DMP) system was selected to obtain a long-lived CL emission. After optimizing the conditions for the CL reaction, the system was applied to the photographic detection, and as little as 250 fmol per spot of CBI-15-mer on a membrane were detected as a visible spot with an instant photographic film.

Base Sequence↗

PubMedDate not supplied

New phenylboronic acid derivatives as enhancers of the luminol-H(2)O(2)-horseradish peroxidase chemiluminescence reaction.

The preparation of three new types of phenylboronic acid derivatives and their evaluation as enhancers on the luminol-H(2)O(2)-horseradish peroxidase (HRP) chemiluminescence (CL) reaction are described. After optimizing the CL reaction conditions, the CL system was applied to the HRP determination. Among the three phenylboronic acid derivatives, i.e. 4-(4, 5-diphenyl-1H-imidazol-2-yl)phenylboronic acid (DPA), 4-[4(or 5)-(4-dimethylaminophenyl)-5(or 4)-phenyl-1H-imidazol-2-yl]phenylboronic acid (DAPA) and 4-[4, 5-di(2-pyridyl)-1H-imidazol-2-yl]phenylboronic acid (DPPA), DPPA was found to be the most potent enhancer. The sensitivity obtained with DPPA was about 180 times higher than that without an enhancer. The detection limit of HRP obtained with DPPA was 0.15 ng/assay (ca. 3.5 fmol), which is comparable to that with 4-iodophenol under the conditions examined. All the phenylboronic acid derivatives examined had the effect of prolonging light emission compared to 4-iodophenol.

Boronic Acids↗

PubMedDate not supplied

Peroxyoxalate chemiluminescent assay for oxidase activities based on detecting enzymatically formed hydrogen peroxide.

A sensitive peroxyoxalate chemiluminescent (PO-CL) assay for activities of oxidases (uricase, choline oxidase, cholesterol oxidase and xanthine oxidase) which catalyse a formation of hydrogen peroxide was developed using 4,4'-oxalyl- bis[(trifluoromethyl-sulphonyl)imino]trimethylene-bis(4- methylmorpholinium)trifluoromethanesulphonate as a chemiluminogenic reagent and 2,4,6,8-tetramorpholinopyrimido[5,4- d]pyrimidine as fluorophore. The standard curve for hydrogen peroxide was linear over the range 1 x 10(-7)-1 x 10(-4) mol/L. Relative standard deviations for oxidase assays were 5.1-12.7% (n = 10). Detection limits were 1 x 10(-3) U/mL for uricase, 5 x 10(-4) U/mL for choline oxidase, 5 x 10(-3) U/mL for cholesterol oxidase and 5 x 10(-4) U/mL xanthine oxidase (sample to blank ratio, 3).

Alcohol Oxidoreductases↗

PubMedDate not supplied

Application of an enhanced luminol chemiluminescence reaction using 4-[4,5-di(2-pyridyl)-1H-imidazol-2-yl]phenylboronic acid to photographic detection of horseradish peroxidase on a membrane.

Photographic detection of horseradish peroxidase (HRP) on a membrane by the luminol-hydrogen peroxide-HRP chemiluminescence reaction using 4-[4,5-di(2-pyridyl)-1H-imidazol-2-yl]phenylboronic acid (DPPA) as an enhancer is described. The method is based on the long-lived chemiluminescence emission obtained by using DPPA. Under the optimum conditions, as little as 0.10 ng (ca. 2.3 fmol) and 0.20 ng (ca. 4.6 fmol) per spot of HRP on a membrane were detected as visible spots with exposure time of 60 and 10 min, respectively, by using an instant photographic film and a camera luminometer. The proposed method was highly sensitive and was successfully applied to the detection of HRP conjugates as an alternative to the colorimetric method using a chromogenic substrate in a commercially available assay kit of Western blotting.

Blotting, Western↗

PubMedDate not supplied

High performance liquid chromatographic analyses of compounds related to nucleic acids by 3-(1-naphthoylamino)- and 3-(1-anthroylamino)-propyl-modified silica gel packing materials.

The development of column packing materials was tried as a contribution to studies related to the further modification of 3-aminopropyl-modified silica gel. Consequently, we prepared two types of gel, 3-(1-naphthoylamino)- and 3-(1-anthroylamino)-propyl-modified silica gels (NAPS and AAPS). Since NAPS and AAPS, each having an aromatic ring, were expected to behave in a separation mode by a pi-pi or hydrophobic interaction, their application to the analyses of the compounds related to nucleic acids was carried out. Adenosine nucleotides were consequently separated by the gels in a single analysis by a single buffer elution with UV detection. NAPS could be also successfully used for the measurement of ATPase activity in fish meat (Sillago japonica). The mobile phase consistently used in the analysis was an aqueous phosphate buffer.

1-Naphthylamine↗

PubMedDate not supplied

High-performance liquid chromatographic separation of penicillamine enantiomers labelled with N-[4-(6-dimethylamino-2-benzofuranyl)phenyl] maleimide on a chiral stationary phase.

Penicillamine enantiomers derivatized with N-[4-(6-dimethylamino-2-benzofuranyl)phenyl]maleimide (DBPM) were separated and determined by high-performance liquid chromatography. A fluorogenic reagent, DBPM easily reacted with D- or L-penicillamine to give each two kinds of strong fluorescent derivatives (D1-, D2-, L1- and L2-DBPM), which could be separated on a Pirkle-type chiral stationary phase using an eluent of 75% aqueous methanol solution containing 0.15 M CH3COONH4 and 0.05 M tetra-n-butylammonium bromide. Two of the peaks (D1- or L1-DBPM), having a shorter retention time than the others, had almost the same retention times (25 min for D1-DBPM and 25.7 min for L1-DBPM). The retention times of the peaks eluted later were 28 min and 31.6 min for D2- and L2-DBPM respectively. Linear calibration curves over the range of 2-50 pmol per injection were obtained for D- and L-penicillamines with a detection limit of 290 and 350 fmol at respectively at a signal-to-noise ratio of 3. Using the proposed method, the absence of contamination of L-penicillamine in a commercially available D-penicillamine preparation (capsule) was confirmed.

Benzofurans↗

PubMedDate not supplied

Importance of serum cholesterol level in development of diabetic autonomic neuropathy.

Changes of cardiac beat-to-beat variation (BBV)--an index of autonomic nerve function--were examined on 2 occasions in an interval of 3.7-5.9 years in 33 diabetics. A total of 204 healthy people, ranging from 11 to 88 years of age, were also subjected to the BBV measurement as controls. In diabetics, a lower rate of change of BBV than the control value was considered as deterioration. Glycohemoglobin was measured every month. The mean of serum cholesterol on the 2 occasions nearest to each time BBV examination was taken for the evaluation of cholesterol level. No significant difference was observed in the control state of blood glucose in terms of glycohemoglobin between the groups with and without BBV deterioration. In 20 subjects with normal cholesterol level, 11 showed a BBV deterioration, and in 13 subjects with abnormal cholesterol level 12 deteriorated in BBV. Therefore, diabetics with normal serum cholesterol level did not seem to exacerbate autonomic neuropathy in terms of BBV.

Adolescent↗

PubMedDate not supplied

Pulmonary adenocarcinoma with micropapillary component: an immunohistochemical study. Case report.

Micropapillary carcinoma has been described in various organs, including the breast, urinary bladder, ovary and lung. We here present a case of pulmonary micropapillary carcinoma in a 72-year-old Japanese man who died of respiratory failure and septic shock, following which autopsy was performed. A mass measuring 2.5 x 2.5 x 2.5 cm was observed in the left lower lobe of the lung. The tumor showed moderately differentiated papillary adenocarcinoma with a focal micropapillary component. Carcinomatous lymphangiosis was also observed in the left lung and metastatic lesions were observed in the bilateral lung, liver, vertebra, muscle layer of the urinary bladder, right adrenal gland, spleen and lymph nodes. The micropapillary component was predominant at some metastatic sites. Immunohistochemically, both the adenocarcinoma and micropapillary components were positive for cytokeratin (CK) 7, CK19, TTF (thyroid transcription factor)-1, carcinoembryonic antigen (CEA) and surfactant apoprotein A (SP-A), and negative for CK20, estrogen receptor, progesterone receptor, uroplakin III, and CA125. The invasive area of the conventional adenocarcinoma component contained a large number of myofibroblasts, whereas the stroma of the micropapillary component contained a small number of myofibroblasts. However, no myofibroblasts were observed in the stroma of the central core of the non-invasive micropapillary carcinoma. Several lymphatic invasions by neoplastic cells were identified in the peripheral area of the micropapillary component using D2-40 antibody. The immunohistochemical profile may be helpful in determining the primary location of the neoplasm containing micropapillary features. Myofibroblasts are present in the stroma of the invasive neoplastic nests in the micropapillary component as well as the conventional adenocarcinoma component, and D2-40 monoclonal antibody may be useful for evaluating the lymphatic invasion of pulmonary micropapillary carcinoma.

Adenocarcinoma, Papillary↗

PubMedDate not supplied

The pupillary light reflex in borderline diabetics.

Computerized IR videopupilography, using an open-loop photic stimulator, was used to compare the reflexes of borderline diabetics with overt non-insulin-dependent diabetics and with age- and sex-matched non-diabetic healthy controls. The patients were all male, aged 41-59 years. Overt non-insulin-dependent diabetics in their 40s had a smaller pupillary area and pupillary diameter prior to photic stimulus compared with borderline diabetics and healthy controls. Overt non-insulin-dependent diabetics in their 50s had a lower pupillary area and pupillary diameter prior to photic stimulus, maximum velocity and acceleration of constriction and maximum velocity of dilation than did borderline diabetics or controls. The only abnormality observed among borderline diabetics compared to controls was a smaller amplitude of constriction in response to light in patients in their 50s. Amplitude of constriction in response to light and velocity of constriction measurements of borderline diabetics, however, were considerably more frequently abnormal than were those of controls. The frequency of abnormal pupillary diameter prior to photic stimulus, maximum velocity of dilation and acceleration of constriction was higher in overt non-insulin-dependent diabetics than in borderline diabetics. The results indicate that borderline diabetics have autonomic neuropathy before the disease becomes overt. The importance of the pupillary light reflex examination is discussed.

Adult↗

PubMedDate not supplied

Myofibroblasts at the tumor border of invasive gastric carcinomas: with special reference to histological type and tumor depth.

In order to elucidate the role of alpha-smooth muscle actin (ASMA)-positive stromal cells, namely myofibroblasts (MFs), in invasive growth of gastric carcinomas (GCs), we examined the number of MFs at the deep border (DB) of GCs. In total 78 invasive GCs (48 intestinal type GCs and 30 diffuse type GCs) were examined immunohistochemically, and analyzed from the view point of tumor depth and histological type. In the intestinal type GCs examined, both the GCs confined to the submucosa and muscularis propria had none or a small number of MFs at the tumor border facing the submucosa (smDB) and muscularis propria (mpDB), respectively; whereas the GCs invading the subserosa had a moderate or large number of MFs at the tumor border facing the subserosa (ssDB) (p<0. 05). Regardless of tumor depth, the diffuse type GCs examined generally had none or a small number of MFs at the smDB, mpDB and ssDB, respectively. There is a possibility that intestinal type GCs invading the subserosa showing tubular and papillary structure may induce myofibroblastic transformation of gastric subserosal stromal fibroblastic cells.

Actins↗