Polymorphisms of glutathione S-transferases (GST) and thymidylate synthase (TS)--novel predictors for response and survival in gastric cancer patients.

To evaluate the predictive value of a panel of gene polymorphisms involved in metabolism of 5-FU and cisplatin on clinical outcome in advanced gastric cancer patients. A total of 52 patients were enrolled in this study. DNA was extracted from paraffin-embedded tumour specimen. Genotypes were determined using PCR-RFLP. Median survival time was 6.0 months (95% CI 3.9;8.1). Overall response rate was 26%. Patients possessing the glutathione S-transferase P1-105 Valine/Valine (GSTP1-105VV) genotype showed a response rate of 67% compared to 21% in patients harbouring at least one GSTP1-105 Isoleucine (GSTP1-105I) allele (P=0.038). GSTP1-105VV patients demonstrated a significant superior median survival time of 15.0 months (95% CI 7.8;22.0) compared to 6.0 months (95% CI 5.1;7.0) in patients with at least one GSTP1-105I allele (P=0.037). Patients possessing a favourable thymidylate synthase (TS) genotype (2R/2R, 2R/3RC, 3RC/3RC) experienced a superior survival time of 10.2 months (95% CI 5.1;15.3) compared to 6.0 months (95% CI 5.0;7.0) in patients with unfavourable TS genotypes (P=0.099). Patients harbouring the GSTP1-105II genotype and one of the unfavourable TS genotypes showed an inferior median survival time of 6.0 months (95% CI 3.9;8.1) compared to 11 months (95% CI 6,23;15,77) in patients with either GSTP1-105VV or a favourable TS genotype (P=0.044). Testing for TS and GSTP1 polymorphisms may allow identification of gastric cancer patients who will benefit from 5-FU/cisplatin chemotherapy, sparing others the side effects of this chemotherapy.

[Growth behaviour of human mononuclear cells derived from bone marrow and cord blood on a collagen carrier for osteogenic regeneration]. / Wachstumsverhalten humaner mononukleärer Zellen aus dem Knochenmark und Nabelschnurblut auf einem Kollagenträger zur osteogenen Regeneration.

UNLABELLED: We investigated the biocompatibility and osteogenetic potency of a porcine collagen I/III carrier in a human bone marrow and cord blood cell culture system. METHODS: Human mesenchymal mononuclear cells were isolated from cord blood and iliac crest bone marrow and cultivated in various cell densities on a semipermeable porcine collagen I/III carrier. After 14 days of in vitro cultivation both cultures were subjected to osteogenic stimulation by dexamethasone, ascorbic acid and beta-glycerol phosphate (DAG) until day 40. Semiquantitative immunochemical evaluation based on osteoblastic and progenitor cell markers was then done. RESULTS: With regard to the minimal local cell density required for growth and osteogenic differentiation, cord blood derived progenitor cells showed lower tolerance in comparison with bone marrow derived cells. For both cell culture systems three-dimensional growth and calcification within the collagen fibres were seen after osteogenic stimulation. CONCLUSION: Human cord blood and bone marrow derived mesenchymal stem cell are capable of differentiating into osteoblasts after incubation with a collagen I/III biomaterial.

Solution structure of an oligonucleotide containing an abasic site: evidence for an unusual deoxyribose conformation.

The antitumor antibiotic bleomycin causes two major lesions in the deoxyribose backbone of DNA: formation of 4'-keto abasic sites and formation of strand breaks with 3'-phosphoglycolate and 5'-phosphate ends. As a model for the 4'-keto abasic site, we have characterized an abasic site (X) in d(CCAAAGXACTGGG).d(CCCAGTACTTTGG) by two-dimensional NMR spectroscopy. A total of 475 NOEs and 101 dihedral angles provided the restraints for molecular modeling. Four unusual NOEs were observed between each anomer of the abasic site and the neighboring bases. In addition, four coupling constants for adjacent protons of the deoxyribose of both the alpha and beta anomers of the abasic site were observed. The modeling suggests that for both anomers the abasic site is extrahelical, without significant distortion of the backbone opposite the lesion. The coupling constants further allowed assignment of an unusual sugar pucker for each anomer. The unique position of the abasic site in our structural model for each anomer is discussed in terms of repair of such lesions in vivo.

Solution structure of Co(III)-bleomycin-OOH bound to a phosphoglycolate lesion containing oligonucleotide: implications for bleomycin-induced double-strand DNA cleavage.

Bleomycin (BLM) is an antitumor antibiotic that is used clinically. Its major cause of cytotoxicity is thought to be related to BLM's ability to cause double-strand (ds) DNA cleavage. A single molecule of BLM appears to cleave both strands of DNA in the presence of its required cofactors Fe(2+) and oxygen without dissociating from the helix. A mechanism for this process has been proposed based on a model structure of the hydroperoxide of Co(III)-BLM (CoBLM) bound sequence-specifically to an intact duplex containing a GTAC site, a hot spot for ds cleavage [Vanderwall, D. E., Lui, S. M., Wu, W., Turner, C. J., Kozarich, J. W., and Stubbe, J. (1997) Chem. Biol. 4, 373-387]. In this paper, we present a structural model for the second cleavage event. Two-dimensional NMR spectroscopy and molecular modeling were carried out to study CoBLM bound to d(CCAAAGXACTGGG).d(CCCAGTACTTTGG), where X represents a 3'-phosphoglycolate lesion next to a 5'-phosphate. Assignments of 729 NOEs, including 51 between the drug and the DNA and 126 within the BLM molecule, have been made. These NOEs in addition to 96 dihedral angle constraints have been used to obtain a well-defined structural model for this complex. The model reveals that the bithiazole tail is partially intercalated between the T19 and the A20 of the duplex and that the metal binding domain is poised for abstraction of the T19 H4' in the minor groove. The modeling further reveals that the predominant conformation of the bithiazole protons is trans. Two cis conformations of these protons are also observed, and ROESY experiments provide evidence for interconversion of all of these forms. The relationship of these observations to the model for ds cleavage is presented.

Thromboembolic disease: comparison of combined CT pulmonary angiography and venography with bilateral leg sonography in 70 patients.

OBJECTIVE: The purpose of this study was to compare combined CT pulmonary angiography and venography with leg sonography for accuracy and relative efficacy in diagnosis of deep venous thrombosis from the popliteal vein to the common femoral vein. SUBJECTS AND METHODS: Seventy consecutive patients with clinically suspected pulmonary embolism underwent both combined CT pulmonary angiography and venography and bilateral leg sonography within 24 hr. CT venograms were analyzed independently in a blinded fashion for quality of venous opacification and patency by two observers. CT venography was compared with sonography for femoropopliteal vein thrombosis, and the final assessment based on multiple subjective and objective clinical and imaging criteria was recorded in three categories: 1, CT venography better than sonography; 2, CT venography equivalent to sonography; and 3, sonography better than CT venography. RESULTS: Sixty-eight patients (97%) had a satisfactory or good quality CT venography examination. Two CT venography studies had false-positive findings due to flow artifacts. Both CT venography and sonography had positive findings for deep venous thrombosis in five patients, and both had negative findings in 63 patients (100% sensitivity, 97% specificity, 100% negative predictive value, and 71% positive predictive value). CT venography was better and more efficacious than sonography (category 1) in 25 patients (36%). CT venography was equivalent to sonography (category 2) in 26 patients (37%), and sonography was better than CT venography (category 3) in 19 patients (27%). CONCLUSION: Compared with sonography, CT venography in addition to CT pulmonary angiography is a relatively accurate method for evaluation of femoropopliteal venous thrombosis. Combined CT pulmonary angiography and CT venography may be more efficacious than sonography or two separate examinations in selected patients.

Functional heterogeneity of isopycnic fractions of rat alveolar macrophages.

Rat alveolar macrophages separated into four isopycnic fractions on Percoll gradients were functionally heterogeneous in bacterial phagocytosis, intracellular killing, superoxide anion release, and lysosomal enzyme activity but not in hydrogen peroxide release. With increasing alveolar macrophage density, phagocytosis, intracellular killing, superoxide anion release, and lysozyme activity increase, and acid hydrolase activity decreases.

Effects of chronic ascorbic acid deficiency on guinea pig lysosomal hydrolase activities.

Previously we have observed increased specific activities of several lysosomal hydrolases in scorbutic guinea pigs and thus the specificity of this effect was examined in guinea pigs marginally deficient in ascorbic acid (AA). Guinea pigs were fed an AA-deficient diet for 2 weeks to deplete body AA pools and then fed a stock diet containing 0.5 mg AA/g diet or the deficient diet plus oral administration of 10 mg AA/day, 1 mg AA/100 g body weight or 0.5 mg AA/100 g body weight each day. Animal were periodically killed during the 12-week experiment and lysosomes isolated from individual livers and analyzed. Serum and brain AA declined when AA was withheld, returned to normal when the stock diet or 10 mg AA were fed but remained at low levels on administation of 1.0 mg or 0.5 mg AA/100 g body weight. Brain norepinephrine followed a similar pattern to brain AA and was opposite to the pattern observed for dopamine. In guinea pigs receiving 1 mg AA/100 g body weight, amine concentrations slowly returned to normal after 8 weeks. Serum hexosaminidase and lysosomal cathepsins A and B were unchanged during the experiment, whereas lysosomal hexosaminidase and acid phosphatase were significantly higher when the experiment was terminated.

Handedness of handedness researchers.

This study tested the hypothesis that laterality and handedness researchers are more likely than other researchers to be left-handed. A questionnaire assessing hand preference was sent to authors of articles which either dealt with handedness (N = 50) or other aspects of laterality (N = 100) or did not concern laterality (N = 50). The proportion of left-handers was highest in the handedness group (12.9%) and lowest in the non-laterality group (5.0%), but this difference did not reach significance.

L-Ascorbic acid and lysosomal acid hydrolase activities of guinea pig liver and brain.

The effects of L-ascorbic acid deficiency on guinea pig hepatic and brain lysosomal hydrolases were examined. In general, hepatic beta-N-acetylhexosaminidase, beta-D-glucoronidase, alpha-D-galactosidase, alpha-D-mannosidase, and acid phosphatase were elevated in scorbutic animals. This appears to be independent of the starved state. Brain beta-D-glucoronidase and acid phosphatase followed a similar pattern to that observed with the liver enzymes, but brain beta-N-acetylhexosaminidase was not affected by L-ascorbic acid decreased the activity of hepatic beta-N-acetylhexosaminiadase was unaffected by dietary treatments although the activity of beta-N-acetylhexosaminidase A tended to increase in the scorbutic animals. Subcellular fractions were obtained from the three groups of animals and the recoveries of protein, beta-N-acetylhexosaminidase, and glucose-6-phosphatase estimated.