Specific tandem GG to TT base substitutions induced by acetaldehyde are due to intra-strand crosslinks between adjacent guanine bases

Acetaldehyde is present in tobacco smoke and automotive exhaust gases, is produced by the oxidation of ethanol, and causes respiratory organ cancers in animals. We show both the types and spectra of acetaldehyde-induced mutations in supF genes in double- and single-stranded shuttle vector plasmids replicated in human cells. Of the 101 mutants obtained from the double-stranded plasmids, 63% had tandem base substitutions, of which the predominant type is GG to TT transversions. Of the 44 mutants obtained from the single-stranded plasmids, 39% had tandem mutations that are of a different type than the double-stranded ones. The GG to TT tandem substitutions could arise from intra-strand crosslinks. Our data indicate that acetaldehyde forms intra- as well as inter-strand crosslinks between adjacent two-guanine bases. Based upon the following observations: XP-A protein binds to acetaldehyde-treated DNA, DNA excision repair-deficient xeroderma pigmentosum (XP) cells were more sensitive to acetaldehyde than the repair-proficient normal cells, and a higher frequency of acetaldehyde-induced mutations of the shuttle vectors was found in XP cells than in normal cells, we propose that the DNA damage caused by acetaldehyde is removed by the nucleotide excision repair pathway. Since treatment with acetaldehyde yields very specific GG to TT tandem base substitutions in DNA, such changes can be used as a probe to identify acetaldehyde as the causal agent in human tumors.     

Immunochemical and biochemical identification of the rice seed protein encoded by cDNA clone A3-12

Previously isolated cDNA clone A3-12 that was expressed in E. coli as the fusion protein with Trp E showed immunoreactivity with the mouse antibody raised against isolated alpha-globulin from rice seed. The N-terminal amino acid sequences determined for the purified alpha-globulin and its tryptic peptides were identical with the deduced amino acid sequence reported, except for two residues at the protein N terminus. An error in the reported sequence was confirmed by re-sequencing the cDNA, the nucleotide sequence for the two N-terminal residues being shown to be CAGCTG and not CACGTG. Thus, the protein encoded by cDNA clone A3-12 was identified to be the major rice seed globulin, alpha-globulin, with an apparent molecular mass of 26kDa.     

Efficacy of microwave coagulation therapy (MCT) in patients with liver tumors

We evaluated the efficacy of microwave coagulation therapy (MCT) in 84 patients with hepatocellular carcinomas (HCC) and 40 with metastatic liver tumors (MLT). The response rates calculated with diagnostic imaging were 92% in HCC and 80% in MLT. The regional recurrence rates were relatively higher in patients with MLT (33%) than in HCC (14%). The average surgical margin in operative MCT group was 11 mm. The cumulative survival rates at three and five years were 63% and 38% in HCC and 43% and 33% in MLT, respectively. The complications were similarly encountered in HCC and MLT (12% versus 13%). When these observations are taken together, MCT is a radical and safe locoregional therapy which can keep an adequate surgical margin and assure long survival.     

High levels of transforming growth factor beta 1 in patients with colorectal cancer: association with disease progression

BACKGROUND & AIMS: Contribution of transforming growth factor beta 1 (TGF-beta 1) to tumor progression has been suggested. However, little is known about the role of TGF-beta 1 in colorectal cancer. Plasma TGF-beta 1 levels and its expression were analyzed in patients with colorectal cancer. METHODS: Plasma TGF-beta 1 levels were measured in 22 patients with colorectal cancer using a TGF-beta 1 enzyme-linked immunosorbent assay. Expression of TGF-beta 1 messenger RNA and immunohistochemical distribution of the protein in colorectal cancer tissues were examined. RESULTS: Plasma TGF-beta 1 levels in patients with colorectal cancer (14.8 +/- 8.4 ng/mL) were significantly higher than in normal controls (1.9 +/- 1.4; n = 22) (P < 0.001). After curative surgical resection, plasma TGF-beta 1 levels decreased in examined patients from 11.9 +/- 6.7 to 3.8 +/- 1.2 ng/mL (P < 0.01). TGF-beta 1 messenger RNA was about 2 1/2 times more abundant in colorectal cancer tissues than in control (P < 0.01). TGF-beta 1 was detected in the cytoplasm of colorectal cancer cells immunohistochemically. Both TGF-beta 1 messenger RNA expression in colorectal adenocarcinoma tissues and its plasma levels were associated with tumor stage of Dukes' classification (P < 0.05). CONCLUSIONS: These results suggest that plasma TGF-beta 1 levels may reflect overexpression of the gene in colon cancer tissues and are associated with disease progression.     

Characterization of phenolic glycolipid-I (PGL-I) like antigen in renal cell carcinoma

BACKGROUND: We investigated whether phenolic glycolipid-I (PGL-I) produced by Mycobacterium leprae (M. leprae), can be used as a marker of renal tumors. METHODS: Using a preparted anti-PGL-I monoclonal antibody (SF-I), anti-PGL-I group antibodies and PGL-I group antigens were detected and PGL-I immunohistologic staining were performed. RESULTS: The titer of anti-PGL-I antibody in patients with renal cell carcinoma was 0.283 +/- 0.103, showing a trend of rising titer of anti-PGL-I group antibody with higher cytologic atypia. When compared by the extent of tumor infiltration, a high antibody titer was observed in stage 4 infiltration group. Regarding the PGL-I group antigen that was detected using SF-1 in the urine of patients with renal cell carcinoma, no relationships between the positive rte of the antigens with common antigenicity to PGL-I and the grade of carcinoma were observed. However, in terms of the extent of tumor infiltration, a trend that the positive rate for PGL-I group antigen increased with the progression of cancer stage was seen. The Western blot assay of the urine of patients with renal cell carcinoma following electrophoresis revealed a change around 40 KD. The immunohistologic stains with SF-1 disclosed a predominance in the proximal kidney tubule. CONCLUSION: Using PGL-I group antigen and PGL-I group antibody as markers of renal cell carcinoma, a rapid and precise measurement may be provided.     

Specific isoprenyl group linked to transducin gamma-subunit is a determinant of its unique signaling properties among G-proteins

Among 11 subtypes of heterotrimeric G-protein gamma-subunit, gamma1 (rod), gamma8 (cone) and gamma11 are modified with farnesyl while the others are modified with geranylgeranyl at the C-terminus. To understand the role of specific isoprenylation (farnesylation) of retinal transducin, we examined how and to what extent the type of isoprenyl group affects transducin-beta gamma (beta1 gamma1) functions such as interactions with membranes, Galpha/receptor, and effectors. To this end, the C-terminal farnesylation signal sequence (CVIS) of gamma1 was replaced by a geranylgeranylation signal (CVIL), and the resultant mutant (S74L) or wild-type (WT) gamma1 was coexpressed with beta1 in the baculovirus-Tn5 insect cell system. Both gamma1WT and gamma1S74L expressed as a beta gamma complex were mixtures modified with farnesyl and geranylgeranyl groups. The ratio of farnesyl to geranylgeranyl in preparations of beta1 gamma1WT and beta1 gamma1S74L purified from the Tn5 cell membrane fraction was about 1:2 and 1:6, respectively. These two forms of recombinant beta1 gamma1 and retinal beta1 gamma1 were different in their abilities to associate with rod outer segment membranes with the following rank order: beta1 gamma1S74L > beta1 gamma1WT > retinal beta1 gamma1. Functionally, beta1 gamma1S74L was the most potent to promote pertussis toxin-catalyzed ADP ribosylation of transducin-alpha (Talpha), to stimulate metarhodopsin II-catalyzed GTPgammaS-binding reaction to Talpha and to modulate adenylyl cyclase and phospholipase C activities. All of the beta1 gamma1 functions absolutely required the isoprenylation of the gamma-subunit. As for the interaction with Goalpha and adenylyl cyclase, predominantly geranylgeranylated beta1 gamma1S74L was less effective than geranylgeranylated beta1 gamma2 purified from bovine brain. These results demonstrate that the properties of Gbeta gamma are strongly affected by the type of functionally indispensable isoprenylation in addition to the amino acid sequence of Ggamma. The relative contribution of the two factors depends on proteins with which Gbeta gamma interacts.     

Application of a novel polypropylene to the insulation of an electric power cable

Cross‐linked polyethylene (XLPE) has been widely adopted as insulating material for high‐voltage power cables up to 500 kV. Further improvement of electrical and thermal properties on insulating material is required in order to increase cable operation efficiency. Therefore, the development of novel insulating material possessing high thermal properties will be necessary. Recent progress of catalysis technology contributes to obtain new polymeric materials which may be applied to electrical insulation. The authors investigated the basic properties of newly developed stereoregular syndiotactic polypropylene (s‐PP) which is synthesized with homogeneous metallocene catalyst. Though recycling of cross‐linked polymers such as conventionally used XLPE may be difficult because of their poor heat deformation, the s‐PP which is not cross‐linked must be suitable for recycling. A series of experiments on its physical and electrical properties gave the following results.

• (1) s‐PP has sufficient flexibility compared with isotactic polypropylene (i‐PP ).
• (2) Both AC and lightning impulse breakdown strength of s‐PP in spite of no cross‐linking are superior to those of XLPE in the temperature range from 25 to 90 °C.
• (3) Degradation by copper of s‐PP is less than that of i‐PP.
• (4) s‐PP/VLDPE blend shows sufficient brittleness temperature for use.

These results suggested that s‐PP should serve as insulating material for power cables at higher‐temperature operation. © 2003 Wiley Periodicals, Inc. Electr Eng Jpn, 146(1): 18–26, 2004; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002//eej.10210