In order to elucidate the mechanisms of mutagenic activation of nitroarenes, we studied the relationships between the mutagenic potency and chemical structure of 2-nitro- and 2,7-dinitro-arenes including nitrated fluorene (Fl), dihydrophenanthrene (DHPh), phenanthrene (Ph), tetrahydropyrene (THPy), dihydropyrene (DHPy) and pyrene (Py) together with 9-NO2-Ph, 1-NO2-Py and 1.3-diNO2-Py. The mutagenicity tests were carried out on Salmonella typhimurium TA98, TA98NR and TA98/1,8-DNP6 in the absence of S9 mix. The order of mutability of mononitro- and dinitro-arenes in TA98 is as given below: 2-NO2-THPy less than 2-NO2-Fl less than 2-NO2-DHPh less than 9-NO2-Ph less than 2-NO2-Ph less than 2-NO2-DHPy less than 1-NO2-Py less than 2-NO2-Py, and 2,7-diNO2-DHPh less than 2,7-diNO2-Fl less than 2,7-diNO2-THPy less than 2,7-diNO2-Ph less than 2,7-diNO2-DHPy less than 2,7-diNO2-Py less than 1,3-diNO2-Py. 9-NO2-Ph and 1-NO2-Py, which have been detected in environmental samples, are not as potent mutagens as 2-nitrated phenanthrene and pyrene, respectively. 2-NO2THPy (37.7 rev/nmole) was a weak mutagen, but 2,7-diNO2-THPy (3197 rev/nmole) was as potent a mutagen as 2,7-diNO2 (3925 rev/nmole). Tetrahydropyrene has a twisted form in its structure. 1,3-diNO2-Py (99660 rev/nmole) was more mutagenic than 2,7-diNO2-Py (37960.0 rev/nmole), and their mutagenicities were correlated with the behavior of the K-band in their UV spectra by the introduction of nitro groups on pyrene.

Biotransformation↗

PubMedDate not supplied

Polymorphism of HSP70 gene is not associated with type 1 (insulin-dependent) diabetes mellitus in Japanese.

Heat shock protein (HSP) 70 is one of the stress-induced proteins and a candidate for islet autoantigen of Type 1 (insulin-dependent) diabetes mellitus. Association of PstI-8.5 kb allele of HSP70 gene with Type 1 diabetes was previously reported in the Caucasian population. Since HSP70 gene is located in the class III region of HLA, the association may be due to the linkage disequilibrium between class II HLA and HSP70 genes. To study whether HSP70 gene is associated with Type 1 diabetes independent of class II HLA genes, we analyzed both HSP70 gene and class II HLA genes in 32 Japanese patients with Type 1 diabetes and 31 control subjects. By Southern blot hybridization with restriction enzyme PstI and HSP70 genomic probe, two allelic bands, 9.0 kb and 8.5 kb, were detected as reported in the Caucasian population. The allele frequencies of 8.5 kb in Type 1 diabetic patients and normal controls were 0.39 and 0.44, respectively. There was no significant difference between the two groups. On the other hand, by PCR-RFLP analysis, DQA1*0301, DQB1*0303 and DQB1*0401 alleles were positively associated with Type 1 diabetes and DQA1*01 was negatively associated with Type 1 diabetes. These data suggest that the polymorphism of HSP70 gene was not associated with Type 1 diabetes in the Japanese population, and that association of PstI-8.5 kb allele with Type 1 diabetes observed in Caucasian population appears to be due to the linkage disequilibrium between this allele and HLA-DR3.

Alleles↗

PubMedDate not supplied

Renal metabolism of 3'-iodohippuryl N(epsilon)-maleoyl-L-lysine (HML)-conjugated Fab fragments.

Renal localization of radiolabeled antibody fragments constitutes a problem in targeted imaging and radiotherapy. Recently, we reported use of a novel radioiodination reagent, 3'-[131I]iodohippuryl N(epsilon)-maleoyl-L-lysine (HML), that liberates m-iodohippuric acid before antibody fragments are incorporated into renal cells. In mice, HML-conjugated Fab demonstrated low renal radioactivity levels from early postinjection times. In this study, renal metabolism of HML-conjugated Fab fragments prepared by different thiolation chemistries and by direct radioiodination were investigated to determine the mechanisms responsible for the low renal radioactivity levels. Fab fragments were thiolated by 2-iminothiolane modification or by reduction of disulfide bonds in the Fab fragments, followed by conjugation with radioiodinated HML to prepare [131I]HML-IT-Fab and [125I]HML-Fab, respectively. In biodistribution studies in mice, both [131I]HML-IT-Fab and [125I]HML-Fab demonstrated significantly lower renal radioactivity levels than those of [125I]Fab. In subcellular distribution studies, [125I]Fab showed migration of radioactivity from the membrane to the lysosomal fraction of the renal cells from 10 to 30 min postinjection. On the other hand, the majority of the radioactivity was detected only in the membrane fraction at the same time points after injection of both [131I]HML-IT-Fab and [125I]HML-Fab. In metabolic studies, while [125I]Fab remained intact at 10 min postinjection, both HML-conjugated Fab fragments generated m-iodohippuric acid as a radiometabolite at the same postinjection time. [131I]HML-IT-Fab registered two radiometabolites (intact [131I]HML-IT-Fab and m-iodohippuric acid), whereas additional radiometabolites were observed with [125I]HML-Fab. This suggested that metabolism of both HML-conjugated Fab fragments would occur in the membrane fractions of the renal cells. The findings of this study reinforced our previous hypothesis that radiochemical design of antibody fragments that liberate radiometabolites that are excreted into the urine by the action of brush border enzymes would constitute a useful strategy to reduce renal radioactivity levels from early postinjection times.

Animals↗

PubMedDate not supplied

Renal metabolism of 111In-DTPA-D-Phe1-octreotide in vivo.

The persistent localization of radioactivity in the kidney after administration of 111In-DTPA-D-Phe1-octreotide impairs the diagnostic accuracy of this radiopharmaceutical. To better understand the mechanisms responsible for the renal radioactivity levels of 111In-DTPA-D-Phe1-octreotide, the renal metabolism of this compound was compared with 111In-DTPA-L-Phe1-octreotide, where the N-terminal D-phenylalanine was replaced with L-phenylalanine to facilitate metabolism. DTPA-D-Phe1-octreotide and DTPA-L-Phe1-octreotide were synthesized by solid-phase methods. Both 111In-DTPA-conjugated octreotide analogues were prepared with radiochemical yields of over 96%, and both remained stable after a 3 h incubation in murine serum at 37 degreesC. When injected into mice, the two 111In-DTPA-conjugated octreotide analogues showed similar radioactivity elimination rates from the blood and accumulation in the kidney with about 60% injected radioactivity being excreted in the urine by 24 h postinjection. Over 85% of the radioactivity in the urine existed as intact peptides for both analogues. Despite the similar renal radioactivity levels, significant differences were observed in the radiolabeled species remaining in the kidney between the two; while 111In-DTPA-L-Phe1-octreotide was rapidly metabolized to the final radiometabolite, 111In-DTPA-L-Phe, the metabolic rate of 111In-DTPA-D-Phe1-octreotide was so slow that various intermediate radiolabeled species were observed. However, both 111In-DTPA-D-Phe and 111In-DTPA-L-Phe remained in the lysosomal compartment of the renal cells as the final radiometabolites for long periods. These findings indicated that although the metabolic stability of 111In-DTPA-D-Phe1-octreotide in the renal cells may be partially involved, the slow elimination rate of the radiometabolite derived from 111In-DTPA-D-Phe1-octreotide from the lysosomal compartment of renal cells would be predominantly attributable to the persistent renal radioactivity levels of 111In-DTPA-D-Phe1-octreotide.

Animals↗

PubMedDate not supplied

Intracellular metabolic fate of radioactivity after injection of technetium-99m-labeled hydrazino nicotinamide derivatized proteins.

Hydrazino nicotinate (HYNIC) has been shown to produce technetium-99m (99mTc)-labeled proteins and peptides of high stability with high specific activities. However, persistent localization of radioactivity was observed in nontarget tissues such as the liver and kidney after administration of [99mTc]HYNIC-labeled proteins and peptides, which compromises the diagnostic accuracy of the radiopharmaceuticals. Since lysosomes are the principal sites of intracellular catabolism of proteins and peptides, 99mTc-HYNIC-labeled galactosyl-neoglycoalbumin (NGA) was prepared using tricine as a co-ligand to investigate the fate of the radiolabel after lysosomal proteolysis in hepatocytes. When injected into mice, over 90% of the injected radioactivity was accumulated in the liver after 10 min injection. At 24 h postinjection, ca. 40% of the injected radioactivity still remained in liver lysosomes. Size-exclusion HPLC analyses of liver homogenates at 24 h postinjection showed a broad radioactivity peak ranging from molecular masses of 0.5-50 kDa. RP-HPLC analyses of liver homogenates suggested the presence of multiple radiolabeled species. However, most of the radioactivity migrated to lower molecular weight fractions on size-exclusion HPLC after treatment of the liver homogenates with sodium triphenylphosphine-3-monosulfonate (TPPMS). The TPPMS-treated liver homogenates showed a major peak at a retention time similar to that of [[99mTc](HYNIC-lysine)(tricine)(TPPMS)] on RP-HPLC. Similar results were obtained with urine and fecal samples. These findings suggested that the chemical bonding between 99mTc and HYNIC remains stable in the lysosomes and following excretion from the body. The persistent localization of radioactivity in the liver could be attributed to the slow elimination rate of the final radiometabolite, [[99mTc](HYNIC-lysine)(tricine)2], from lysosomes, and subsequent dissociation of one of the tricine co-ligands in the low pH environment of the lysosomes in the absence of excess co-ligands, followed by binding proteins present in the organelles. The findings in this study also suggested that the development of appropriate co-ligands capable of preserving stable bonding with the Tc center is essential to reduce the residence time of radioactivity in nontarget tissues after administration of [99mTc]HYNIC-labeled proteins and peptides.

Albumins↗

PubMedDate not supplied

Gelsolin gene therapy by retrovirus producer cells for human bladder cancer in nude mice.

Gelsolin, a regulator of the actin cytoskeleton, has been shown previously to act as a tumor suppressor in vitro and in vivo when introduced into certain cancer cell lines. To investigate the in vivo efficacy of gene therapy with the gelsolin gene, we inoculated nude mice with human urinary bladder cancer cells (UMUC-2 or DAB-1) and tested the effects of adding either retroviral DNA constructs containing gelsolin cDNA or retrovirus producer cells that produce the same retroviral constructs at high levels. The addition of retroviral gelsolin cDNA constructs did not inhibit tumor growth; however, this form of gene therapy, in which retrovirus producer cells were introduced, resulted in marked and reproducible tumor growth inhibition and prolonged survival time in the majority of animals tested. These findings demonstrate the potential for treating human urinary bladder carcinomas with the gelsolin gene.

3T3 Cells↗

PubMedDate not supplied

Assessing prosocial message effectiveness: effects of message quality, production quality, and persuasiveness.

The purpose of this study was to determine whether the effectiveness of prosocial messages is compromised by poor design. A receiver-oriented content analysis (N = 246) was used to assess college students' perceptions of the message quality, production quality, and persuasiveness of advertisements and prosocial advertisements regarding alcohol. After providing background information, respondents rated a series of video clips on a variety of criteria guided by the Message Interpretation Process (MIP) model. Results indicated that prosocial advertisements were rated as higher in quality than were commercial advertisements overall and on logic-based criteria, but prosocial advertisements nevertheless had weaker relationships to viewers' beliefs and reported behaviors relevant to drinking alcohol. Heavier drinkers rated commercial advertisements more positively than did lighter/nondrinkers. They were less skeptical of persuasive messages and rated prosocial advertisements lower in effectiveness and commercial advertisements higher in effectiveness.

Adult↗

PubMedDate not supplied

A rat model of dilated cardiomyopathy to investigate partial left ventriculectomy.

BACKGROUND: This study is the first to assess a small animal model of dilated cardiomyopathy (DCM) for evaluation of partial left ventriculectomy. METHOD: Eighteen Dahl salt-sensitive (DS) rats were divided into three groups. Six rats were fed an 8% high-salt diet from the age of 7 weeks (Group 1), and similarly six rats from 8 weeks (Group 2) and six from 9 weeks (Group 3). Blood pressure (BP) was measured by the tail-cuff method and left ventricular (LV) dimensions by echocardiography. RESULTS: In Groups 1 and 2, systolic BP rose and reached 200 mmHg by the 10th to 11th week, when all rats died within a week without signs of heart failure. However, in Group 3, systolic BP gradually rose to 196+/-15 mmHg (mean +/- SD) at the age of 14 weeks, when LV end-diastolic diameter (EDD) was 6.2+/-0.4 mm (control 5.1+/-0.7 mm) and LV fractional shortening (FS) was 77+/-3% (control 68+/-3%). At the age of 25 to 30 weeks, all rats in Group 3 showed signs of congestive heart failure, systolic BP remained high, EDD markedly increased (8.7+/-0.6 mm), and LVFS decreased (38.9+/-8.1%). From this stage, rats survived for 13.7+/-5.9 days. We employed the Group 3 model for our pilot PLV study. Eight rats had PLV with a beating heart by plicating the LV area between the papillary muscle bases. Two rats died perioperatively but the rest survived (60% survival 3 weeks after PLV). Postoperatively, the rats' LVEDD decreased and FS improved significantly. CONCLUSIONS: Using DS rats, we developed a DCM model for investigating PLV. The model may contribute to scientific investigation of PLV.

Animals↗