Biomedical subjects

J Zhao

Publications and source records attributed to J Zhao.

At least 19 recordsLinked to original sources

PubMed2001

Temperature-induced decoupling of phycobilisomes from reaction centers.

Temperature-induced decoupling of phycobilisomes (PBSs) from the reaction centers in the PBS-thylakoid membrane complexes was observed at 0 degrees C. The fluorescence yields of photosystem (PS) I and PSII decreased and that of PBSs increased with selective excitation of PBSs at 0 degrees C, while the yield of PBSs decreased and those of the two photosystems increased with selective excitation of chlorophyll a at room temperature (RT). It indicated that the decoupling of PBSs from the two photosystems led to changes of energy transfer efficiencies, which can be explained by partial detachment of PBSs from thylakoid membrane. The temperature-dependent processes were reversible, i.e. with temperature going up to RT, the complexes could restore to the functionally coupled state with a time constant about 30 s. Based on these results, it could be deduced that PBSs should be in parallel connection with the two photosystems.

Betaine↗

PubMed2001

In vitro degradation and release profiles for poly-dl-lactide-poly(ethylene glycol) microspheres containing human serum albumin.

Poly-dl-lactide-poly(ethylene glycol) (PELA) block copolymers containing same the content (10%) of polyethylene glycol (PEG) were synthesized with five different molecular weight of PEG by ring-opening polymerization. PELA microspheres containing human serum albumin (HSA) were elaborated by solvent extraction method based on the formation of double w/o/w emulsion. In vitro matrix degradation and protein release of these microspheres were performed in phosphate-buffered saline (PBS) (154 mM, pH 7.43). The degradation profiles were characterized by measuring the loss of microspheres mass, the decrease of polymer intrinsic viscosity, the decrease of pH value of degradation medium, the reduction of polymer number-average molecular weight (M(n)) and the change of molecular weight polydispersity (M(w)/M(n)). The release profiles were investigated from the measurement of protein presented in the release medium at various intervals. It showed that the matrix degradation and protein release profiles were highly polymer-dependent. The extent of burst release in the initial protein release increased with the decrease of molecular weight of PELA copolymer. It is suggested that these matrix polymers may be optimized as carriers in protein (antigen) delivery system for different purposes.

Biocompatible Materials↗

PubMed2001

Actin bound to the heterogeneous nuclear ribonucleoprotein hrp36 is associated with Balbiani ring mRNA from the gene to polysomes.

In the salivary glands of the dipteran Chironomus tentans, a specific messenger ribonucleoprotein (mRNP) particle, the Balbiani ring (BR) granule, can be visualized during its assembly on the gene and during its nucleocytoplasmic transport. We now show with immunoelectron microscopy that actin becomes associated with the BR particle concomitantly with transcription and is present in the particle in the nucleoplasm. DNase I affinity chromatography experiments with extracts from tissue culture cells indicate that both nuclear and cytoplasmic actin are bound to the heterogeneous RNP (hnRNP) protein hrp36, but not to the hnRNP proteins hrp23 and hrp45. The interaction is likely to be direct as purified actin binds to recombinant hrp36 in vitro. Furthermore, it is demonstrated by cross linking that nuclear as well as cytoplasmic actin are bound to hrp36 in vivo. It is known that hrp36 is added cotranscriptionally along the BR mRNA molecule and accompanies the RNA through the nuclear pores and into polysomes. We conclude that actin is likely to be bound to the BR transcript via hrp36 during the transfer of the mRNA from the gene all the way into polysomes.

Actins↗

PubMed2001

Pulmonary hypoplasia in mice lacking tumor necrosis factor-alpha converting enzyme indicates an indispensable role for cell surface protein shedding during embryonic lung branching morphogenesis.

Many membrane-bound protein precursors, including cytokines and growth factors, are proteolytically shed to yield soluble intercellular regulatory ligands. The responsible protease, tumor necrosis factor-alpha converting enzyme (TACE/ADAM-17), is a transmembrane metalloprotease-disintegrin that cleaves multiple cell surface proteins, although it was initially identified for the enzymatic release of tumor necrosis factor-alpha (TNF-alpha). Mammalian lung growth and development are tightly controlled by cytokines and peptide growth factors. However, the biological function of the cell shedding mechanism during lung organogenesis is not understood. We therefore evaluated the role of TACE as a "sheddase" during lung morphogenesis by analyzing the developmental phenotypes of lungs in mice with an inactive TACE gene in both in vivo and ex vivo organ explant culture. Neonatal TACE-deficient mice had visible respiratory distress and their lungs failed to form normal saccular structures. These newborn mutant lungs had fewer peripheral epithelial sacs with deficient septation and thick-walled mesenchyme, resulting in reduced surface for gas exchange. At the canalicular stage of E16.5, the lungs of TACE mutant mice were impaired in branching morphogenesis, inhibited in epithelial cell proliferation and differentiation, and delayed in vasculogenesis. Embryonic TACE knockout mouse lungs (E12) branched poorly compared to wild-type lungs, when placed into serumless organ culture. Gene expression of both surfactant protein-C and aquaporin-5 were inhibited in cultured TACE-mutant embryonic lungs, indicating defects in both branching and peripheral epithelial cytodifferentiation in the absence of TACE protein. Furthermore, both the hypoplastic phenotype and the delayed cytodifferentiation in TACE-deficient lungs were rescued by exogenous addition of soluble stimulatory factors including either TNF-alpha or epidermal growth factor in embryonic lung culture. Thus, the impaired lung branching and maturation without TACE suggest a broad role for TACE in the processing of multiple membrane-anchored proteins, one or more of which is essential for normal lung morphogenesis. Taken together, our data indicate that the TACE-mediated proteolytic mechanism which enzymatically releases membrane-tethered proteins plays an indispensable role in lung morphogenesis, and its inactivation leads to abnormal lung development.

ADAM Proteins↗

PubMed2001

Two phases of chromatin decondensation during dedifferentiation of plant cells: distinction between competence for cell fate switch and a commitment for S phase.

Cellular dedifferentiation is the major process underlying totipotency, regeneration, and formation of new stem cell lineages in multicellular organisms. In animals it is often associated with carcinogenesis. Here, we used tobacco protoplasts (plant cells devoid of cell wall) to study changes in chromatin structure in the course of dedifferentiation of mesophyll cells. Using flow cytometry and micrococcal nuclease analyses, we identified two phases of chromatin decondensation prior to entry of cells into S phase. The first phase takes place in the course of protoplast isolation, following treatment with cell wall degrading enzymes, whereas the second occurs only after protoplasts are induced with phytohormones to re-enter the cell cycle. In the absence of hormonal application, protoplasts undergo cycles of chromatin condensation/decondensation and die. The ubiquitin proteolytic system was found indispensable for protoplast progression into S phase, being required for the second but not the first phase of chromatin decondensation. The emerging model suggests that cellular dedifferentiation proceeds by two functionally distinct phases of chromatin decondensation: the first is a transitory phase that confers competence for cell fate switch, which is followed, under appropriate conditions, by a second proteasome-dependent phase representing a commitment for the mitotic cycle. These findings might have implications for a wide range of dedifferentiation-driven cellular processes in higher eukaryotes.

Base Sequence↗

PubMed2001

Adenovirus-mediated transfer of inducible caspases: a novel "death switch" gene therapeutic approach to prostate cancer.

In patients with localized prostate cancer, radical prostatectomy and radiation therapy, although effective in controlling localized disease, are often associated with significant side effects attributable to injury of adjacent tissues. Moreover, patients with metastatic disease eventually fail systemic hormonal or chemotherapy because of the development of progressive, refractory disease. In this study, we evaluated the safety and efficacy of a novel suicide gene therapy that could potentially spare normal tissue while bypassing molecular mechanisms of apoptosis resistance by using chemically inducible effector caspases to trigger apoptosis in prostate cancer cells. Initially, we compared the ability of a panel of inducible Fas signaling intermediates to kill human and murine prostate cancer cell lines. On the basis of the superior killing by downstream caspase-1 and caspase-3, replication-deficient adenoviral vectors expressing conditional caspase-1 (Ad-G/iCasp1) or caspase-3 (Ad-G/iCasp3), regulated by nontoxic, lipid-permeable, chemical inducers of dimerization (CID), were constructed. Upon vector transduction followed by CID administration, aggregation and activation of these recombinant caspases occur, leading to rapid apoptosis. In vitro, both human (LNCaP and PC-3) and murine (TRAMP-C2 and TRAMP-C2G) prostate cancer cell lines were efficiently transduced and killed in a CID-dependent fashion. In vivo, direct injection of Ad-G/iCasp1 into s.c. TRAMP-C2 tumors caused focal but extensive apoptosis without evidence for a bystander effect at the maximal viral dose (i.e., 2.5 x 10(10) viral particles/25 microl) in host animals that also received CID compared with control animals. Treatment with Ad-G/iCasp1 plus CID resulted in a transient, yet significant, reduction both in tumor growth and volume compared with tumors treated with vector but not CID (P < 0.035) or vector-diluent plus CID (P < 0.022), both of which grew more rapidly. These results demonstrate that CID-regulated, caspase-based suicide gene therapy is safe and can inhibit the growth of experimental prostate cancer in vitro and in vivo through potent induction of apoptosis, providing a rationale for further development.

Adenocarcinoma↗

PubMed2001

Functional characterization of rat gp600/megalin promoter: combination of proximal Sp1 site and JCV repeat is important in rat gp600/megalin promoter activation.

Gp600/megalin is an endocytic receptor belonging to the low-density lipoprotein receptor family. Up or down regulation of this protein were observed in certain disease states. To understand the mechanisms that control gp600/megalin gene expression, we cloned and functionally characterized a 738-bp fragment of the 5'-flanking region of rat gp600/megalin gene. A transcription start site was mapped to 33 bp downstream of TAGAAA sequence (TATA-like box). Multiple transcription factor binding sites were identified. Serial 5' deletions and transient transfection assays showed that the deletion fragment containing the Sp1 site proximal to the TATA-like box and a JCV repeat retained 80% of the promoter activity. Individual mutations of the proximal Sp1 site and JCV repeat reduced the promoter activity by 60 and 34% respectively. Double mutations of the proximal Sp1 site and JCV repeat produced a dramatic 80% reduction in the promoter activity. However, deletions and mutations or double mutations of other transcription factor binding sites in the promoter region had a minor effect on the promoter activity. These results indicate that the combination of proximal Sp1 site and the JCV repeat are necessary for activation of gp600/megalin expression. Moreover, Sp1 and Sp3 proteins interacted with the proximal and the distal Sp1 sites in the nuclear extracts of gp600/megalin expressing cell lines. TCF site seems to be involved in negative regulation of this promoter but no nuclear protein(s) were found to bind to this site. In addition, Ap2 site responsible for 28% promoter activity is able to bind two dominant unknown nuclear proteins. This functional characterization of the regulation of gp600/megalin gene is likely to advance the knowledge of the regulation of this gene in health and disease.

Animals↗

PubMed2001

Novel structures and properties of gold nanowires.

The structures of free-standing gold nanowires are studied by using molecular-dynamics-based genetic algorithm simulations. Helical and multiwalled cylindrical structures are found for the thinner nanowires, while bulk-like fcc structures eventually form in the thicker nanowires up to 3 nm in diameter. This noncrystalline-crystalline transition starts from the core region of nanowires. The vibrational, electronic, and transport properties of nanowires are investigated based on the optimal structures. Bulklike behaviors are found for the vibrational and electronic properties of the nanowires with fcc crystalline structure. The conductance of nanowires generally increases with wire diameter and depends on the wire structure.

Journal Article↗

PubMed2001

Focal adhesion kinase activates Stat1 in integrin-mediated cell migration and adhesion.

Recent studies suggest that focal adhesion kinase (FAK) is important for cell migration. We now suggest a mechanism by which FAK activates the signal transducer and activator of transcription (STAT) pathway, regulating cell adhesion and migration. In particular, we observe that FAK is capable of activating Stat1, but not Stat3. Co-immunoprecipitation and in vitro binding assays demonstrate that Stat1 is transiently and directly associated with FAK during cell adhesion, and Stat1 is activated in this process. FAK with a C-terminal deletion (FAKDeltaC14) completely abolishes this interaction, indicating this association is dependent on the C-terminal domain of FAK, which is required for FAK localization at focal contacts. Moreover, Stat1 activation during cell adhesion is diminished in FAK-deficient cells, correlating with decreased migration in these cells. Finally, we show that depletion of Stat1 results in an enhancement of cell adhesion and a decrease in cell migration. Thus, our results have demonstrated, for the first time, a critical signaling pathway from integrin/FAK to Stat1 that reduces cell adhesion and promotes cell migration.

Blotting, Western↗

PubMed2001

Down-regulation of promoter 1.3 activity of the human aromatase gene in breast tissue by zinc-finger protein, snail (SnaH).

Aromatase (estrogen synthetase) is expressed in breast cancer tissue, and in situ expression of the enzyme stimulates breast cancer growth. Promoter I.3 is one of the major promoters that control the expression of aromatase in breast cancer tissue. Using the yeast one-hybrid approach to screen a human breast tissue hybrid cDNA expression library, we found that the zinc-finger transcriptional factor Snail (SnaH) interacted with a regulatory region near promoter I.3 of the human aromatase gene. DNA mobility shift assays and mutation analyses using recombinant SnaH protein expressed in Escherichia coli have revealed that this protein interacts with a segment, 5'-CTGATGAAGT-3', which is between 66 and 76 bp upstream from the transcriptional start site of promoter I.3. Using mammalian cell transfection experiments, SnaH was found to act as a repressor of promoter I.3 activity. Site-directed mutagenesis experiments have revealed that the NH2-terminal SNAG domain is important for the repressor activity of SnaH. To demonstrate the inhibitory activity against aromatase expression, a stable SnaH-expressing MDA-MB-231 breast cancer cell line was generated, and the aromatase RNA messages in the SnaH-transfected cell line were found to be 30% of those in the vector-transfected cell line. Reverse transcription-PCR analysis on RNAs isolated from 12 cell lines has confirmed that SnaH is expressed at a higher level in normal breast epithelial cell and stromal fibroblast cell lines than in breast cancer cell lines. In addition, SnaH mRNA was detected in only 16 of 55 breast cancer specimens. On the other hand, aromatase mRNA was detected in 54 of the 55 specimens. Our results indicate that SnaH acts as a repressor that down-regulates the expression of aromatase in normal breast tissue by suppressing the function of promoter I.3. A reduction of the expression of SnaH in breast cancer tissue further suggests a cancer-protective role for this protein in normal breast tissue.

Amino Acid Sequence↗

PubMed2001

Quantitative changes in spinal canal dimensions using interbody distraction for spondylolisthesis.

STUDY DESIGN: An experimental study was performed using cadaveric lumbar spines to evaluate the effect of anteriorly or laterally placed interbody distraction implants on the alteration of spinal canal and neuroforaminal dimensions. OBJECTIVES: To quantify changes in the spinal canal and neuroforaminal dimensions using interbody fusion devices inserted at various configurations in cadaveric lumbar spines exhibiting degenerative spondylolisthesis. SUMMARY OF BACKGROUND DATA: Although several clinical studies have demonstrated successful treatment of degenerative spondylolisthesis with anterior interbody fusion, no study has shown the role of interbody distraction in improving lumbar spinal canal and foraminal stenosis. METHODS: Five fresh cadaver lumbar spines exhibiting a degenerative spondylolisthesis or retrospondylolisthesis were used for the study. Computed tomography scans of each specimen and a silicon mold of the left intervertebral foramens were repeated in a consistent manner after pure compressive load (150 lb) was applied to simulate physiologic load (intact case), after two BAK (Sulzer SpineTech Inc., Minneapolis, MN) distraction plugs were anteriorly inserted into the intervertebral disc space (anterior distraction cases), and after one long BAK cage was laterally inserted from the left side (lateral distraction case). The cross-sectional area of the spinal canal was measured from computed tomography images using National Institutes of Health image software (Bethesda, MD). The spinal canal volume was calculated using the cross-sectional area and total scan thickness. Left intervertebral foraminal volumes were calculated from the weight of the silicon mold injected into the foramen. Descriptive statistics and a Student's t test were used to detect statistical differences in the spinal canal and neuroforaminal volumes before and after interbody distraction. RESULTS: The cross-sectional canal area was significantly increased after anterior distraction (35.11%) and lateral distraction (33.14%). The spinal canal volume was markedly increased with anterior distraction (19.92%) and lateral distraction (21.96%). Left foraminal volume was also enhanced by 40.25% for anterior distraction and 41.03% for lateral distraction. CONCLUSIONS: Interbody distraction either by anteriorly inserted plugs or laterally inserted threaded cagescan immediately improve the narrowed canal area and increase spinal canal, as well as foraminal volume for lumbar degenerative spondylolisthesis or retro- spondylolisthesis.

Anthropometry↗

PubMed2001

Insulin-like growth factor-I (IGF-I) stimulates the development of cultured rat pre-antral follicles.

The aim of the present study is to investigate the effect of insulin-like growth factor (IGF-I) on the development of cultured rat pre-antral follicles. Pre-antral follicles with a diameter between 140 and 160 microm are mechanically isolated from ovaries of 10-day-old rats and cultured in groups for 6 days in FSH and insulin-containing serum-free medium in the absence or presence of IGF-I at concentrations of 1, 10, and 100 ng/ml, respectively. DNA content of the follicles before and after culture is measured to evaluate whether a possible growth is due to proliferation of follicular cells and the ultrastructure of the cultured follicles is studied with transmission electron microscopy to evaluate the quality of follicles cultured under different conditions. Furthermore, reverse transcriptase polymerase chain reaction (RT-PCR) is used to assess the gene expression of IGF-I and type I IGF receptor in pre-antral follicles. When follicles were cultured in medium containing IGF-I at a concentration of 1 and 10 ng/ml, the increase in follicle diameter after a 6-day culture differs significantly from that in the absence of IGF-I. The ultrastructure of follicles cultured in IGF-I-containing medium is better sustained when compared to that of follicles cultured in the absence of IGF-I. Especially, theca cells, which are undergoing degeneration under controlled culture condition, demonstrate normal cellular ultrastructure with some characteristics of steroid-secreting cells in the presence of IGF-I. Moreover, in oocytes of follicles cultured in the presence of IGF-I, cortical granules are observed distributed along the ooplasma membrane whereas cortical granules are hardly present in follicles cultured without IGF-I. With RT-PCR, the presence of the mRNA of IGF-I and type I IGF receptor is demonstrated in both the somatic follicular cells and the oocytes. Taken together, these results suggest that IGF-I, in the presence of follicle-stimulating hormone (FSH), enhances rat pre-antral follicle development in vitro and supports the morphology of cultured pre-antral follicles. The stimulatory effect of exogenous IGF-I on the development of pre-antral follicle together with the gene expression of both IGF-I and type I IGF receptor in pre-antral follicles suggests the involvement of IGF-I in early folliculogenesis and its actions are likely mediated by the putative type I IGF receptor, via both endocrine and paracrine/autocrine pathways.

Animals↗

PubMed2001

Determination of synthesized alpha-vitamin E by micellar electrokinetic chromatography.

We developed a micellar electrokinetic chromatography method (MEKC) for the direct determination of the content of synthesized alpha-vitamin E. It was found that under the optimum separation conditions 7 mM borate + 14 mM phosphate + 15 mM sodium dodecyl sulfate (SDS) + 10 mM sodium cholate (NaCh) + 8% acetonitrile (pH 9.2) with UV detection wavelength at 214 nm, 16 kV constant voltage, and 26 degrees C constant temperature, alpha-vitamin E and its isomers can be baseline separated and alpha-vitamin E was quantitatively analyzed. In addition, the sample recovery, the limit of detection and the repeatability of the method were investigated. The influence of various parameters on the separation such as SDS concentration, NaCh concentration, buffer pH and acetonitrile percentage were also discussed.

Acetonitriles↗

PubMed2001

Gains of 12q13-14 and overexpression of mdm2 are frequent findings in intimal sarcomas of the pulmonary artery.

The characterization of clinical, histopathological, immunohistochemical, and genetic features of intimal sarcomas arising in the pulmonary artery is presented in this study. Four resected lungs, one endarterectomy specimen and three biopsies from eight patients (four males and four females; median age 41 years) suffering from intimal sarcomas of the pulmonary artery using conventional stains, immunohistochemistry, and comparative genomic hybridization (CGH) were analyzed. The predominant clinical presentation was dyspnea (all eight patients) and febrile pulmonary disease (six of eight). Signs of embolic lung disease were present in all patients. One patient died postoperatively, six patients died of disease 8-35 months after presentation, and one patient was alive 6 months after surgery. Histopathological examination of the submitted material showed spindle cell, partially myxoid and pleomorphic sarcomas. Metastases were histologically confirmed in three patients (lung, pleura, and skull). Immunohistochemically, vimentin was strongly expressed in all tumors. Focal positivity was observed for alpha smooth muscle actin, CD117, CD68, p53, and bcl2. No reaction could be obtained for endothelial markers. The proliferation index Ki-67 was between 5% and 80%. Six examined tumors were positive for mdm2. In the CGH analysis, gains and amplifications in the 12q13-14 region were found in six of eight tumors (75%). Other, less consistent alterations, were losses on 3p, 3q, 4q, 9p, 11q, 13q, Xp, and Xq, gains on 7p, 17p, and 17q, and amplifications on 4q, 5p, 6p, and 11q. Intimal sarcomas of the pulmonary artery are tumors with an unfavorable prognosis and poorly differentiated morphology. A majority of tumors show a consistent genetic alteration (gains and amplifications in the 12q13-14 region) and overexpression of mdm2, implicating the mdm2/p53 pathway as a possible mechanism in the tumor pathogenesis.

Actins↗

PubMed2001

Application of concentrated TiO2 sols for gamma-ray radiation dosimetry.

Upon exposure to gamma-radiation, a concentrated TiO2 sol changes from colorless to deep blue with an absorption maximum at 540 nm. The absorption has been assigned to trapped electrons or Ti3+ states in the solid matrix based on its spectroscopic similarity to the samples irradiated with UV light. Unlike the conduction-band electrons generated from direct excitation by UV radiation, the origin of the trapped electrons during gamma-ray irradiation may be traced to a series of reducing species produced by the high energy electrons, which in turn, are the direct result of gamma-irradiation. As the absorption intensity is linearly related to the duration of exposure to gamma-radiation, it may have an application in gamma-ray dosimetry. The sensitivity of its dosage response has been found to be influenced by the semiconductor particle concentration and the dispersing solvent.

Journal Article↗

PubMed2001

Spectroscopy study on the photochromism of Schiff bases N,N'-bis(salicylidene)-1,2-diaminoethane and N,N'-bis(salicylidene)-1,6-hexanediamine.

The photochromism of Schiff bases N,N'-bis(salicylidene)-1,2-diaminoethane (BSE) and N,N'-bis(salicylidene)-1,6-cyclohexanediamine (BSH) was studied by steady-state and time-dependent fluorescence, UV Vis absorption spectroscopy and theoretical chemistry calculations. The experimental results show that BSH can perform the photochromism easier than BSE, may be due to the molecular topology difference.

Ethylenediamines↗

PubMed2001

Individual-specific risk factors for anorexia nervosa: a pilot study using a discordant sister-pair design.

BACKGROUND: The aim of this pilot study was to examine which unique factors (genetic and environmental) increase the risk for developing anorexia nervosa by using a case-control design of discordant sister pairs. METHODS: Forty-five sister-pairs, one of whom had anorexia nervosa and the other did not, were recruited. Both sisters completed the Oxford Risk Factor Interview for Eating Disorders and measures for eating disorder traits, and sib-pair differences. Blood or cheek cell samples were taken for genetic analysis. Statistical power of the genetic analysis of discordant same-sex siblings was calculated using a specially written program, DISCORD. RESULTS: The sisters with anorexia nervosa differed from their healthy sisters in terms of personal vulnerability traits and exposure to high parental expectations and sexual abuse. Factors within the dieting risk domain did not differ. However, there was evidence of poor feeding in childhood. No difference in the distribution of genotypes or alleles of the DRD4, COMT, the 5HT2A and 5HT2C receptor genes was detected. These results are preliminary because our calculations indicate that there is insufficient power to detect the expected effect on risk with this sample size. CONCLUSIONS: A combination of intrinsic and extrinsic factors increases the risk of developing anorexia nervosa. It would, therefore, be informative to undertake a larger study to examine in more detail the unique genetic and environmental factors that are associated with various forms of eating disorders.

Adolescent↗

PubMed2001

Detection and quantification of depurinated benzo[a]pyrene-adducted DNA bases in the urine of cigarette smokers and women exposed to household coal smoke.

Polycyclic aromatic hydrocarbons (PAH) are metabolized to electrophiles that can bind to DNA bases and destabilize the N-glycosyl bond, causing rapid depurination of the adducted bases. Recent studies support depurination of DNA as a mechanism central to the genesis of H-ras mutations in PAH-treated mouse skin. Depurinating adducts account for 71% of all DNA adducts formed in mouse skin treated with benzo[a]pyrene (BP). This study analyzed urine of cigarette smokers, coal smoke-exposed women, and nonexposed controls for the presence and quantities of the depurinated BP-adducted DNA bases, 7-(benzo[a]pyren-6-yl)guanine (BP-6-N7Gua) and 7-(benzo[a]pyren-6-yl)adenine (BP-6-N7Ade). Since these adducted bases originate from reaction of the BP radical cation with double-stranded DNA and not with RNA or denatured DNA, their presence in urine is indicative of DNA damage. Urine samples were fractionated by a combination of SepPak extraction and reverse-phase HPLC, and then analyzed by tandem mass spectrometry and capillary electrophoresis with laser-induced fluorescence. BP-adducted bases were detected in the urine from three of seven cigarette smokers and three of seven women exposed to coal smoke, but were not detected in urine from the 13 control subjects. Concentrations were estimated to be 60-340 and 0.1-0.6 fmol/mg of creatinine equivalent of urine for coal smoke-exposed women (maximum possible BP intake of ca. 23 000 ng/day) and cigarette smokers (BP intake of ca. 800 ng/day), respectively, exhibiting a sensitive response to BP exposures. BP-6-N7Gua was present at ca. 20-300 times the concentration of BP-6-N7Ade in the urine of coal smoke-exposed women, but was not detected in the urine of cigarette smokers. This difference may be due to the remarkably different BP exposures experienced by the two groups of PAH-exposed individuals. These results justify more extensive studies of depurinated BP-adducted DNA bases as potential biomarkers of PAH-associated cancer risk.

Adult↗