Improvement of long-term prognosis in patients with ovarian cancers by adjuvant sizofiran immunotherapy: a prospective randomized controlled study

The effect of immunotherapy using sizofiran (SPG) on the prognosis of patients with ovarian cancers was prospectively studied in a total of 68 patients, who were randomly assigned to either a cisplatin, adriamycin and cyclophosphamide (PAC) therapy group or a PAC plus SPG combination therapy group.The survival rate was significantly higher in patients with stage Ic, II or III cancers treated with the PAC plus SPG combination, compared with the patients treated with PAC alone. In the SPG-receiving patients with stage Ic or more advanced cancers who were treated with four cycles or more of PAC, the outcome was improved (Cox-Mantel, p=0.074; generalized Kruskal-Wallis, p=0.032). Similar improvement was also observed in the patients with non-serous adenocarcinomas (Cox-Mantel, p-0.076; generalized Krukal-Wallis, p=0.045). No side effects attributable to SPG were recorded.The present results suggest that the use of SPG in combination with long-term chemotherapy improves the postoperative prognosis in ovarian cancer patients.Abbreviations SPG sizofiran     

Characterization of four monosialo and a novel disialo Asn N-glycosides from the urine of a patient with aspartylglycosaminuria

We previously reported for the first time two Japanese patients with aspartylglycosaminuria (AGU). A novel disialo Asn N-glycoside (AG-5) has been isolated from the urine of one of the patients in addition to four known monosialo Asn N-glycosides (AG-1 to AG-4) by gel filtration and anion exchange chromatography in this study. Final purification of AG-5 was achieved by an electrochemical chromatographic method, high performance liquid chromatography with pulsed amperometric detector (HPLC-PAD). The yield of AG-5 was approximately 1 mg l^–1 urine. The chemical structures of AG-1 to AG-5 were characterized by gas-liquid chromatography, a permethylation study, fast atom bombardment-mass spectrometry (FAB-MS), and nuclear magnetic resonance (NMR). Based on the structural analysis, AG-5 had the following novel structure: NeuAc2 8NeuAc2 3Gal1 4GlCNAc1 Asn.     

Involvement of blood-group-B-active trisaccharides in Ca2+-dependent cell-cell adhesion in the Xenopus blastula

 Despite their wide distribution in various organisms, no physiological roles have been proposed for the human blood-group-ABO (ABH)-active trisaccharides. Here we show that monoclonal antibodies against human blood-group-B-active trisaccharides (B-substance) completely block the Ca²⁺-dependent cell-cell adhesion system of frog (Xenopus laevis) embryonic cells. Synthetic B-substance or B-active glycopeptides also disrupt the Ca²⁺ -dependent cell-cell adhesion. These results suggest that blood-group-B-active substances play a role in cell-cell adhesion. Blood-group-B-active substances were found as glycoproteins and as glycosphingolipids. In order to identify B-active glycoproteins active in cell-cell adhesion, we purified B-active membrane glycoproteins by two-dimensional electrophoresis and found that they are 45- to 58-kDa proteins with pI(s) ranging from 4.0 to 5.3. They are glycosylphosphatidyl inositol (GPI) anchored. Amino acid sequence analysis showed that the purified B-active GPI-anchored proteins are homologues of soluble Xenopus cortical granule lectins (CGL). The results suggest that the B-active membrane glycoproteins are GPI-anchored forms of the lectin and are directly involved in frog Ca ²⁺-dependent cell-cell adhesion. Received: 16 September 1997 / Accepted 19 November 1997     

Selective determination of haloperidol in human serum: surface ionization mass spectrometry and gas chromatography with surface ionization detection

Surface ionization organic mass spectrometry (SIOMS) has been performed on the clinically important drug haloperidol using quadrupole mass spectrometry in which the thermal ion source has a rhenium oxide emitter. The surface ionization (SI) mass spectrum is presented, interpreted in a purely empirical way by means of evidence from previous investigations, and then compared to results from conventional electron impact (EI) ionization. An approach to detection of this drug in serum by gas chromatography (GC) with a surface ionization detector (SID) and GC-SIOMS is described. This approach demonstrates that (a) haloperidol is efficiently surface-ionized, giving a unique SI mass spectrum, (b) experimental results rationalize the combined sensitivity and selectivity of the GC-SID for the examined drug, (c) the detection limit for haloperidol in serum is 1.1 ng/ml (S/N = 3) by GC-SID (the coefficients of variation of the assay are generally low, i.e. below 8.5%) and (d) the GC-SIOMS coupling can be used for sensitive and selective detection of haloperidol in serum.     

Peripheral Tγ lymphocyte population in head and neck cancer

Summary Peripheral T lymphocytes were measured in head and neck cancer patients and controls. The percentage was significantly higher in the 59 cancer patients than in the 46 normal controls (P<0.001). The 12 patients with recurrent disease had elevated percentages of T lymphocytes compared with the untreated group (n=31; P<0.05) and the treated, disease-free group (n=16; P<0.05). Moreover, the percentage of T lymphocytes was significantly higher in the 31 patients with regional lymph node metastasis than in the node-negative group (n=28; P<0.05). In a total of 37 patients with squamous cell carcinoma histologically graded I, II, and III, the absolute counts and percentages of T lymphocytes in the grade I group (n=13) showed significant decreases compared with those in the grade III group (P<0.05; n=6). Moreover, postoperative serial determinations of the percentage of T lymphocytes in the 14 treated, disease-free patients revealed a gradual decrease of T lymphocytes, whereas the five patients with recurrent disease had a tendency to increases in the percentage of T lymphocytes. Abbreviations used in this paper: SMF, sodium metrizoate-Ficoll; PBS, phosphate-buffered saline; SRBC, sheep erythrocytes; FCS, fetal calf serum; Hepes, hydroxyethylpiperazin elthene-sulfonic acid; E, erythrocyte; EAC, erythrocyte-antibody-complement; K, antibody-dependent killer; NK, natural killer     

The induction of reversible and irreversible chromosome decondensation by protein synthesis inhibition during meiotic maturation of mouse oocytes

We investigated the effects of puromycin on mouse oocyte chromosomes during meiotic maturation in vitro. Puromycin treatment for 6 hr at 100 μg/ml almost completely, but reversibly, suppressed [³⁵S]methionine incorporation into oocyte protein at all stages of maturation tested. Nevertheless, oocytes treated at the germinal vesicle stage underwent germinal vesicle breakdown (GVBD) and chromosome condensation. These oocytes completed nuclear maturation to metaphase II (MII) if the inhibitor was withdrawn. Prolonged (24-hr) treatment, however, caused the chromsomes to degenerate. The chromosomes of oocytes treated shortly after GVBD for 6 hr remained condensed, but the oocytes failed to form a polar body. However, 24-hr treatment caused the chromosomes to decondense to form an interphase nucleus. Oocytes treated near MI for 6 hr gave off a polar body during the treatment, and their chromosomes decondensed to form a nucleus, which remained as long as the treatment was continued. However, if the puromycin was withdrawn, the chromosomes recondensed to a state morphologically similar to that at MII. Thus, the chromosome decondensation induced by protein synthesis inhibition at MI was reversible. Oocytes treated at MII, several hours after first polar body formation, also underwent chromosome decondensation to form a nucleus. In the continuous presence of puromycin, the chromosomes remained decondensed, but neither DNA synthesis nor mitosis occurred. However, following puromycin withdrawal, these occytes synthesised DNA and underwent mitosis. Thus, protein synthesis inhibition at MII, by parthenogenetically activating the oocytes, caused irreversible chromosome decondensation. Based on these observations, we discussed the roles of protein synthesis in the regulation of oocyte chromosome behaviour during meiotic maturation.     

The relationship of structure of glucoamylase and glucose oxidase to antigenicity

Glucoamylase and glucose oxidase fromAspergillus niger have been purified to homogeneity by chromatography on DEAE-cellulose and the purified enzymes have been used to investigate structural and antigenicity relationships. In structure, glucoamylase and glucose oxidase are glycoproteins containing 14% and 16% carbohydrate. Earlier methylation and reductive -elimination results have shown that glucoamylase has an unusual arrangement of carbohydrate residues, with 20 single mannose units and 25 di-, tri-, or tetrasaccharide chains of mannose, glucose, and galactose, all attached O-glycosidically to serine and threonine residues of the protein moiety. The antigenicity of the glucoamylase has now been found to reside predominantly in the types and arrangement of the carbohydrate chains. Glucose oxidase contains mannose, galactose, and glucosamine in the N-acetyl form in the native enzyme, but the complete structure of the carbohydrate chains has not yet been determined. The antigenicity of this enzyme does not reside in the carbohydrate units, but rather in the polypeptide chains of the two subunits of the enzyme. Glucose oxidase can be dissociated into subunits by mercaptoethanol and sodium dodecyl sulfate treatment, while glucoamylase cannot be dissociated, but undergoes only an unfolding of the polypeptide chain under these conditions. The subunits of glucose oxidase do not react with the anti-glucose oxidase antibodies, but the unfolded molecule and peptide fragments produced from glucoamylase by cyanogen bromide cleavage do react with antiglucoamylase antibodies.     

Amino acids and peptides. X. Synthesis of two dotriacontapeptides corresponding to C-terminal sequence 30–61 (fragment α) of human liver metallothionein II and their heavy metal binding properties

Two kinds of dotriacontapeptides corresponding to C-terminal sequence 30–61 of human liver metallothionein, both containing 11 cysteine residues, were synthesized by the conventional fragment condensation method employing the HF deprotection or MSA deprotection method at the final step. Their heavy metal (Zn, Cd, or Cu) binding activity was examined, and it was found that their heavy metal binding properties were quite similar to those of native human liver thionein.