Biomedical subjects

L Li

Publications and source records attributed to L Li.

At least 613 records · Page 34Linked to original sources

PubMed2000

The A5.1 allele of the major histocompatibility complex class I chain-related gene A is associated with psoriasis vulgaris in Chinese.

BACKGROUND: Recent studies have demonstrated an association of a polymorphic (GCT)n triplet repeat in the transmembrane (TM) region of the major histocompatibility complex (MHC) class I chain-related gene A (MICA), one of the MHC class I chain-related (MIC) family members, with some autoimmune diseases, including Behçet's disease, acute anterior uveitis, Takayasu's arteritis and others. OBJECTIVES: The aim of this study was to examine whether the MICA gene is associated with psoriasis vulgaris (PS) in Chinese. PATIENTS AND METHODS: The (GCT)n polymorphism of the MICA gene was investigated in 200 healthy Chinese of Han origin and 300 patients with PS by polymerase chain reaction amplification and denaturing polyacrylamide gel electrophoresis. RESULTS: Five alleles, namely A4, A5, A6, A9 and A5.1 were found in both groups. Comparison of the data from both groups revealed that the A5.1 allele was present at a significantly higher frequency in the patient group (41.5%) than in the control group (23.0%) (Pc < 0.0001, Pc means the probability of a comparison with the control group). The frequency of A5.1-positive cases was also significantly increased in the patient group (68.0%) as compared with the controls (38.0%) (Pc < 0.0001). Furthermore, the carrier frequency of A5.1-positive was significantly increased in psoriatic patients with a positive family history and with early onset as compared with sporadic cases (Pc = 0.0005) and with late onset PS (Pc = 0.002). CONCLUSIONS: These results suggest that the MICA gene may be associated with the development of PS in Chinese.

Adolescent↗

PubMed2000

Proteolytic components of serum IgG preparations.

Chemical catalysis, an effector mechanism utilized by fully assembled antibodies, can also be mediated by the isolated antibody subunits. Because trace amounts of free light chains (L chains) are present in IgG preparations, a detailed study was undertaken to identify the constituents responsible for the polyreactive proteolytic activity of IgG purified from human sera, determined as the extent of cleavage of the model peptide substrate Pro-Phe-Arg-methylcoumarinamide. Two proteolytic species with approximate mass of 50 kD and 150 kD were separated by repetitive gel filtration in a denaturing solvent (6 M guanidine hydrochloride). The activity of the renatured 50-kD fraction (in fluorescence units/microg protein) was more than 45-fold greater than of the 150-kD fraction. Both fractions lost the activity following immunoadsorption on immobilized anti-IgG antibody. Fab fragments prepared from the 150-kD IgG fraction retained the activity. Reducing and non-reducing SDS-electrophoresis suggested the 50-kD fraction isolated from the IgG preparations to be a mixture of heavy chain (H chain) monomers and disulphide bonded L chain dimers. Electrophoretically homogeneous monomers of 50-kD H chains and 25-kD L chains were prepared by gel filtration of reduced and alkylated IgG from seven human subjects. Each of the alkylated L chain preparations displayed the proteolytic activity. The activity in alkylated H chains was undetectable or only marginally greater than the background values. L chain dimers appear to be the major species responsible for the polyreactive proteolytic activity of serum IgG preparations, with a smaller contribution furnished by tetrameric IgG.

Antibodies, Catalytic↗

PubMed2000

Accumulation of amyloid-beta protein in exocrine glands of transgenic mice overexpressing a carboxyl terminal portion of amyloid protein precursor.

Amyloid-beta protein (Abeta) and its precursor (betaPP) play important roles in the pathogenesis of Alzheimer disease and inclusion-body myositis. In humans, Abeta deposits are found in brain, skeletal muscle, and skin. Therefore, we have investigated possible Abeta deposits in multiple tissues of two transgenic mouse lines overexpressing the signal plus Abeta-bearing 99-amino acid carboxyl terminal sequences of betaPP under the control of a cytomegalovirus enhancer/beta-actin promoter. One of the lines developed Abeta-immunoreactive intracellular deposits consistently in the pancreas and lacrimal gland, and occasionally in gastric, DeSteno's, and lingual glands. Although the Abeta deposits increased during ageing and degenerative changes of the tissues were observed, little or no extracellular Abeta deposits were observed up to the age of 25 months. These lines of transgenic mice are useful for studying the molecular mechanisms of development and clearance of intracellular Abeta deposits.

Aging↗

PubMed2000

A novel large plasmid carrying multiple beta-lactam resistance genes isolated from a Klebsiella pneumoniae strain.

Klebsiella pneumoniae clinical isolates were selected according to the results of antibiotic susceptibility tests. Most of them were resistant to multiple antibiotics, including ampicillin, ceftazidime, cefotaxime and aminoglycosides. Large plasmids were observed in these Kl. pneumoniae strains by pulsed-field gel electrophoresis with S1 nuclease digestion. The Kl. pneumoniae strains investigated produced one to two extrachromosomal bands with a mobility corresponding to 97 approximately 145 kbp linear DNA molecules. A 100 kbp plasmid, designated pK1, was observed in the multiply resistant strain K250. pK1 had sequences homologous to both the TEM-1 and the aphD probe which were associated with beta-lactam and aminoglycoside resistance. pK1 was transformed into Escherichia coli strain DH5alpha and was found to confer resistance to ampicillin, ceftazidime, cefotaxime and kanamycin. A 8 kbp BamHI DNA fragment of pK1 that carried the ampicillin resistance gene (minimum inhibitory concentration > 1000 microgram ml-1) was cloned into the BamHI site of pACYC184. Sequence determination showed that this cloned fragment carried a TEM-1 gene. These findings suggest that pK1 is novel in that it appears to carry genes for resistance to ampicillin, cefotaxime and ceftazidime, as well as kanamycin.

Aminoglycosides↗

PubMed2000

Effect of interleukin-7 on the in vitro development and maturation of monocyte derived human dendritic cells.

We have compared the cell phenotype and functional properties of monocyte/macrophage derived dendritic cells (DCs) obtained by culture of human adherent peripheral blood mononuclear cells (PBMCs) in medium containing granulocyte macrophage colony stimulating factor (GM-CSF) either alone (GM-CSF-DCs), or in combination with interleukin (IL)-4 (IL4-DCs) or IL-7 (IL7-DCs). The cell surface phenotype of GM-CSF-DCs and IL-7-DCs was characterized by a high expression of major histocompatibility complex (MHC) class I and II, CD80, CD86 and CD40. In contrast to 'classical' IL-4-DCs, these two types of DCs expressed CD14 and a CD21-like molecule detected by two out of four CD21-specific monoclonal antibodies (MoAb) tested. The same pattern of reactivity with CD21 specific antibodies was observed in freshly isolated adherent PBMCs but not in B lymphocytes. This reactivity was upregulated by IL-7 in a dose dependent manner. Lipopolysaccharide (LPS) treatment induced the upregulation of CD40, CD80, CD86 and the T-cell stimulatory capacity in IL-4-DCs and, to a lesser extent, in the IL-7-DCs whereas GM-CSF-DCs responded very poorly to such treatment. Our data indicate that, together with GM-CSF, the IL-7 drives macrophage precursors to a differentiation stage that is close to but distinct from the phenotype of IL-4-DCs. Comparison of DC development in the presence of IL-7 or IL-4 may help in dissecting signalling pathways that regulate the expression of functionally relevant DC markers.

Antigens, CD↗

PubMed2000

A novel transmembrane serine/threonine protein kinase gene from a rifamycin SV-producing amycolatopsis mediterranei U32.

Genomic DNA sequencing in the vicinity of methylmalonyl-CoA mutase gene (mutAB) from a rifamycin SV-producing Amycolatopsis mediterranei U32 allowed us to clone, sequence, and identify a gene encoding a novel serine/threonine protein kinase (amk). The sequence contains a complete ORF of 1821 base pairs encoding a predicted protein of 606 amino acids in length. The N-terminal domain of the protein shows significant homology to the catalytic domain of other protein kinases from both prokaryotic and eukaryotic sources. It also contains all the structural features that are highly conserved in active protein kinases, including the Gly-X-Gly-X-X-Gly motif of ATP-binding and the essential amino acids known to be important for the recognition of the correct hydroxyamino acid in serine/threonine protein kinase. This protein kinase gene was expressed in Escherichia coli and was shown to have the ability of autophosphorylation. The autophosphorylated site was found to be the threonine at position 164 by labeled phosphoamino acid analysis and site-directed mutagenesis. The C-terminal half of protein kinase was found to contain strong transmembrane structures by PhoA fusion protein analysis, suggesting that Amk protein kinase is a transmembrane protein. A Southern hybridization experiment showed that this type of protein kinase is distributed ubiquitously and might play significant physiological roles in the various species of streptomycetes. However, overexpression of amk gene in Streptomyces cinnamonensis showed no effect on methylmalonyl-CoA mutase activity, monensin production and the hyphae morphology. Although its biological role is still unknown, Amk protein kinase is the first transmembrane serine/threonine protein kinase described for genus Amycolatopsis.

Actinomycetales↗

PubMed2000

Frequency and significance of antibodies to chromatin in autoimmune hepatitis type I.

AIMS AND METHODS: To assess the frequency and clinical significance of antibodies to chromatin (ACA) in autoimmune hepatitis (AIH), 36 Japanese patients with AIH type I were studied for serum reactivity with chromatin by using an ELISA. RESULTS: Antibodies to chromatin were detected in 19 of 36 patients with AIH type I. There was a significantly higher frequency of ACA in patients with AIH type I than in patients with primary biliary cirrhosis, chronic hepatitis C and B (52.8 vs 13.2, 5.4 and 6.7%, respectively; P<0.01). None of the 19 healthy subjects had positive reactions. Sixteen of 19 patients with seropositive sera (44.4%) had reactivities with other nuclear antigens (recombinant nucleoproteins U1RNP-A, U1RNP-70; recombinant ribonucleoprotein complexes SSA/Ro 52K, SSA/Ro 60K; recombinant centromere Cenp-B; dsDNA and histones). Adsorption with double-stranded DNA (dsDNA) and histones could not remove the majority of antichromatin reactivity as 81.9% of the antibody reactivity still remained. In five sera samples from AIH type I patients positive for anti-dsDNA and antihistones, the antibody activities for dsDNA and histones were inhibited after the absorbtion of sera with chromatin. The patterns of antinuclear antibodies (ANA) detected by using indirect immunofluorescence were similar between patients with and without ACA. Patients with ACA had significantly high serum levels of gamma-globulin and immunoglobulin G. The ACA titers dropped significantly after corticosteroid treatment (P< 0.01). CONCLUSIONS: Antibodies to chromatin are frequently present in patients with AIH type I and they are one of the dominant autoantibodies associated with ANA reactivity in AIH type I. Antibodies to chromatin cannot be used to characterize distinct clinical subgroups of AIH type I.

Adrenal Cortex Hormones↗

PubMed2000

Rotavirus infection in children in Japan.

Currently, a high morbidity of rotavirus diarrhea has been seen in children in developed and developing countries. Improvement of the vaccines is necessary in order to reduce the burden of diarrhea caused by rotavirus. A survey of rotavirus infection from diarrheal stool specimens in children of seven regions in Japan was conducted from 1984 to 1999. The present study discusses the survey results and reviews the national and international data of more than 23 papers and congress proceedings about rotavirus infection in Japan. We analyze the prevalence of rotavirus infection in acute diarrheal in- and outpatients, the distribution of rotavirus G-serotypes and surveillance data for seasonality and age groups in Japan. The data indicated that rotavirus is the most important cause of diarrhea in Japan among young children, with the prevalence ranging from approximately 9.7 to 88%. The most common rotavirus strains belonged to serotype G1, specifically since 1993. Serotypes G2, G3 and G4 had also been documented to be predominantly based in the area and year before 1992. However, untypeable rotavirus strains had been found each year, with a prevalence up to 56.7% which suggests that rare serotypes (except G1-4) or new serotypes might exist. Unexpectedly, in Tokyo and Sapporo from 1998 to 1999, G9 was found to be the first most prevailing serotype with a high prevalence of 52.9 and 71.4%, respectively. Despite these data from different geographic areas, the year under investigation was relatively clear in respect to seasonality, with a peak of rotavirus activity in late winter (February) through early spring (March). Age distribution had also characterized that the infection was predominant among children aged 1-2 years of age, although it was also common in children of 2-3 years. In addition, mixed infection with bacteria was documented.

Adolescent↗

PubMed2000

TT virus infections among blood donors in Iceland: prevalence, genotypes, and lack of relationship to serum ALT levels.

BACKGROUND: The TT virus (TTV) is a newly identified blood-borne virus. Its association with disease is still unknown, and screening of blood donors has not been implemented. Several genotypes of the TTV have been identified. STUDY DESIGN AND METHODS: Three hundred seventy healthy blood donors were randomly selected and tested for TTV by the PCR method. Sequencing of a part of the genome was performed to identify various genotypes of the virus. ALT levels were determined in both infected and uninfected individuals. RESULTS: The TT virus (TTV), was detected in the sera of 23 (6.2%) of 370 healthy Icelandic blood donors; this prevalence is lower than that reported in Japan but higher than that in Scotland. The virus was found in all groups over the age of 19. Sequencing and phylogenetic analysis of 202 bp from open reading frame 1 demonstrated genotypes 1b and 2b 2c and genotype 4 isolates, with the latter bearing 89-percent nucleotide homology with other genotype 4 sequences deposited at GenBank. One sample showed a mixed genotype 1b/2c infection. Serum ALT levels were within normal limits in all infected individuals. CONCLUSION: The TTV carrier state does not cause significant liver injury.

Adolescent↗

PubMed2000

Posterior sagittal approach: megasigmoid resection and anal reconstruction for severe constipation and fecal incontinence after anoplasty.

PURPOSE: The aim of this study was to present the technique of megasigmoid resection and anal reconstruction by complete posterior sagittal approach for the children with severe constipation and fecal incontinence after anoplasty. METHODS: Six patients (age, 2 to 18 years) born with imperforate anus and originally treated with perineal anoplasty suffered from intractable constipation and fecal incontinence. Contrast enema showed massive dilated and aperistaltic rectosigmoid colon with fecal impaction. Resection of the dilated bowel and anal reconstruction were completely performed by posterior sagittal approach. RESULTS: The mean operating time was 205 minutes (range, 125 to 265 minutes) and the average length of resected colon was 23.3 cm (range, 10 to 40 cm). There were no intraoperative or postoperative complications. By 2 to 4 months after the operation, all patients obtained voluntary bowel movement. On follow-up at 6 to 24 months postoperative, no patient had constipation or required use of the laxatives again. Four of 6 patients suffered from grade 1 soiling, and the other 2 had grade greater than 1 soiling. None had urinary retention or incontinence after the procedure. CONCLUSION: Resection of dilated rectosigmoid colon and anal reconstruction for the patients with severe constipation and fecal incontinence after anoplasty can be performed successfully using a posterior sagittal approach.

Adolescent↗

PubMed2000

Congenital biliary dilatation.

Congenital biliary dilatation is commonly associated with pancreaticobiliary malunion. Its etiology remains unknown. With the advent of accurate cholangiography, combined abnormalities of the intrahepatic duct, common channel, and pancreatic duct are being identified more frequently in patients with congenital biliary dilatation. Early diagnosis followed by cyst excision is the treatment of choice, even in asymptomatic cases. The treatment of intrahepatic and intrapancreatic ductal diseases such as intrahepatic duct dilatation and stone debris in the intrahepatic duct and common channel are also discussed. The value of intraoperative endoscopy as an adjunct to cyst excision for the prevention of postoperative complications is explained.

Bile Ducts, Intrahepatic↗

PubMed2000

Antiplatelet and antithrombotic effects of the diterpene spiramine Q from Spiraea japonica var. incisa.

Spiramine Q, a diterpene, was isolated from a Chinese herbal plant Spiraea japonica var. incisa Yu. Born's and Wan HY's methods were used to investigate effects of spiramine Q on rabbit platelet aggregation and serotonin release, respectively. Its antithrombotic effect in mice was also evaluated by Myers' method. Spiramine Q selectively inhibited arachidonic acid-induced platelet aggregation in vitro or ex vivo, and decreased serotonin secretion from rabbit platelets. Spiramine Q (5 mg/kg) decreased the mouse mortality caused by injection of 80 mg/kg arachidonic acid in the tail vein. The results suggested that spiramine Q showed potent antiplatelet and antithrombotic activites.

Animals↗

PubMed2000

Chemical constituents of two Chinese Magnoliaceae plants, Tsoongiodendron odorum and Manglietiastrum sinicum, and their inhibition of platelet aggregation.

Phytochemical investigations of Tsoongiodendron odorum and Manglietiastrum sinicum, both Magnoliaceae, led to the isolation of twenty compounds in total. Among them, one was a new sesquiterpene, 11-O-oleoyl-beta-eudesmol (2), and another, 1-(3,4-dimethoxypheny)-4-(3,4-methylenedioxyphenyl)-2,3-dimethy lbutane (12) was isolated as a natural product for the first time. Moreover, 13C-NMR spectral data of isoguaiacin (16) are reported here for the first time. Structure elucidations for compounds reported here were mainly based on their spectral data. The ethanolic extracts of T. odorum and M. sinicum, and six pure compounds, 4(15)-eudesmen-11-ol (beta-eudesmol) (1), 1 beta-hydroxy-4(15),11(13)-eudesmadien-12,6 alpha-olide (reynosin) (3), 3,11(13)-eudesmadien-12,6 alpha-olide (alpha-cyclocostunolide) (5), erythro-1-(4-hydroxy-3-methoxyphenyl)-4-(3,4-methylenedioxyphenyl)-2,3- dimethylbutane (11), nectandrin-B (18), and syringaresinol (19), displayed considerable inhibition against platelet aggregation induced by AA, by ADP, or by PAF.

Humans↗

PubMed2000

Protection against ozone-induced pulmonary inflammation and cell death by endotoxin pretreatment in mice: role of HO-1.

Ozone is a ubiquitous air pollutant that can cause acute pulmonary inflammation and cell injury and may contribute to the exacerbation of chronic pulmonary diseases. The molecular mechanisms of ozone-induced cell injury, as well as protective mechanisms against ozone-injury, are not well understood. Since ozone is a reactive oxidant, and heme oxygenase-1 (HO-1) is an antioxidant enzyme induced by many oxidative stimuli, we hypothesized that HO-1 is one of the protective mechanisms against ozone-induced cell injury, as well as pulmonary inflammation. In the current study, C57Bl/6 mice were pretreated with a low level of endotoxin (lipopolysaccharide, LPS) (0.5 mg/kg) to induce HO-1, and 16 h later were exposed to 1 ppm ozone for 3 h. Endotoxin pretreatment caused a significant protection against ozone-induced pulmonary inflammation and cell injury in bronchoalveolar lavage (BAL) cells. The protection by endotoxin pretreatment against ozone-induced inflammation and necrosis in BAL cells was abolished by the cotreatment with a heme oxygenase inhibitor, tin protoporphyrin IX dichloride (SnPP), suggesting that HO-1 is responsible for the protection against ozone-induced pulmonary inflammation and BAL cell necrosis. Therefore, since HO-1 is induced following ozone exposure, HO-1 may contribute to the development of cellular adaptation to chronic ozone exposure.

Animals↗

PubMed2000

XPD/ERCC2 polymorphisms and risk of head and neck cancer: a case-control analysis.

DNA repair capacity is central in maintaining normal cellular functions. Variants of several DNA repair genes,including the nucleotide excision repair gene XPD, have been described recently. Because we previously reported that patients with squamous cell carcinoma of the head and neck (SCCHN) had lower DNA repair capacity than healthy controls, we hypothesized that inherited polymorphisms of XPD may contribute to genetic susceptibility to SCCHN, a tobacco-related cancer. To test this hypothesis, we conducted a hospital-based case-control study of 189 SCCHN patients and 496 cancer-free controls who were frequency-matched on age, gender and smoking status. All subjects were non-Hispanic whites. Two XPD polymorphisms (C22541A and A35931C) were typed using the restriction enzymes TfiI and PstI, respectively. Multivariate logistic regression analysis was performed to calculate adjusted odds ratios (ORs) and 95% confidence intervals (CIs). In the controls, the frequencies of the variant 22541A and 35931C alleles were 44.7% and 33.8%, respectively. The frequency of the 22541A homozygous genotype (22541AA) was lower in cases (15.9%) than in controls (20.4%) but was not associated with risk (adjusted OR = 0.90; 95% CI = 0.52-1. 56) for SCCHN. The frequency of the 35931C homozygous genotype (35931CC) was higher in cases (16.4%) than in controls (11.5%) and associated with a borderline increased risk (adjusted OR = 1.55; 95% CI = 0.96-2.52) for SCCHN. The risk was higher in older subjects (OR = 2.22; 95% CI = 1.03-4.80), current smokers (OR = 1.83; 95% CI = 0.79-4.27) and current drinkers (OR = 2.59; 95% CI = 1.25-5.34) in the stratification analysis. These results suggest a gene-environment interaction, but this did not reach statistical significance. The findings are limited due to the relatively small numbers in the subgroups and need to be verified by further investigations.

Adult↗

PubMed2000

Effect of a tyrosine 155 to phenylalanine mutation of protein kinase cdelta on the proliferative and tumorigenic properties of NIH 3T3 fibroblasts.

Tyrosine phosphorylation has emerged as an important mechanism in the regulation of enzyme function. In this paper, we describe a mutant of PKCdelta altered at a single tyrosine residue which has the opposite effect compared with wild-type PKCdelta on the growth characteristics of NIH 3T3 cells. Overexpression of wild-type PKCdelta results in a decreased growth rate and a lower cell density at confluency. On the other hand, overexpression of PKCdelta with a mutation from tyrosine to phenylalanine at position 155 results in a significantly higher rate of growth and a higher density at confluency compared with vector controls. Moreover, these cells are able to grow in soft agar and to form tumors in nude mice. In contrast to kinase negative PKC constructs, this mutant maintains in vitro kinase activity and shows a subcellular localization and a translocation pattern that are similar to those of the wild-type PKCdelta. Whether the altered biological effect is due to the missing phosphorylation on tyrosine or the mutation from tyrosine to phenylalanine per se remains under investigation.

3T3 Cells↗

PubMed2000

A prospective study of limiting longstanding illness in early adulthood.

BACKGROUND: Chronic illness and disability are of increasing public health importance but little is known about the lifetime influences involved in their onset and progression. We aim to (i) establish whether an individual's rating of limiting illness is stable over a 10-year period from age 23 to 33; (ii) assess the relationship between childhood and adult disability; and (iii) identify lifecourse influences on limiting illness in early adulthood. METHODS: Data were from the 1958 British birth cohort, including the original birth survey and follow-ups at ages 7, 11, 16, 23 and 33 years. Limiting longstanding illness was the outcome at both ages 23 and 33. Potential predictors included childhood health and physical development, socioeconomic conditions in early life and adulthood, and behavioural factors. We estimated the effect of potential explanatory factors using logistic regression, in both univariate and multivariate analyses, separately for limiting illness at 23 and 33 years. RESULTS: Prevalence of limiting illness increased from 5.1% (men) and 4.1% (women) at age 23 to 6% for both sexes at age 33. Risk of limiting illness at age 33 was greater for those reporting an illness at age 23 (29.4%, compared with 4.7% of those without illness), though the majority (66%) of 33-year limiting illnesses had no previous record at age 23 or for childhood. Multivariate analysis of limiting illness at age 23 confirmed the high risk for those with childhood disability and also established two further major predictors, namely, injury (adjusted odds ratio [OR] = 1.42, 95% CI: 1.09-1.86) and intermediate socio-emotional status (adjusted OR = 1.73, 95% CI: 1.29-2.31). Additional risks were identified for limiting illness at age 33, including: (i) injury in the preceding 10 years (adjusted OR = 1.55, 95% CI : 1.18-2.04); (ii) body mass index (BMI), for which the relationship was non-linear, with elevated risks for the underweight (adjusted OR = 1.53, 95% CI: 1.03-2.26) and overweight (OR = 1.28, 95% CI: 0.87-1.89); (iii) childhood disadvantage at either or both ages 7 and 11 (adjusted OR = 1.53, 95% CI : 1.07-2.17); and (iv) height at age 7, with a significant non-linear relationship (the adjusted OR for height less than 15th percentile was 1.43 and for height more than the 85th percentile, 1.30). CONCLUSIONS: Both childhood and adult factors predict limiting illness in early adulthood. Childhood is important because some adult illnesses originate in early life, and also because childhood environment influences the risk of adult limiting illness several years later. Our findings suggest that studies seeking to understand the causes of limiting illness, that currently tend to focus exclusively on contemporary factors, need also to consider the contribution of environment in early life.

Adaptation, Psychological↗

PubMed2000

Toxicogenomics-based discrimination of toxic mechanism in HepG2 human hepatoma cells.

The rapid discovery of sequence information from the Human Genome Project has exponentially increased the amount of data that can be retrieved from biomedical experiments. Gene expression profiling, through the use of microarray technology, is rapidly contributing to an improved understanding of global, coordinated cellular events in a variety of paradigms. In the field of toxicology, the potential application of toxicogenomics to indicate the toxicity of unknown compounds has been suggested but remains largely unsubstantiated to date. A major supposition of toxicogenomics is that global changes in the expression of individual mRNAs (i.e., the transcriptional responses of cells to toxicants) will be sufficiently distinct, robust, and reproducible to allow discrimination of toxicants from different classes. Definitive demonstration is still lacking for such specific "genetic fingerprints," as opposed to nonspecific general stress responses that may be indistinguishable between compounds and therefore not suitable as probes of toxic mechanisms. The present studies demonstrate a general application of toxicogenomics that distinguishes two mechanistically unrelated classes of toxicants (cytotoxic anti-inflammatory drugs and DNA-damaging agents) based solely upon a cluster-type analysis of genes differentially induced or repressed in cultured cells during exposure to these compounds. Initial comparisons of the expression patterns for 100 toxic compounds, using all approximately 250 genes on a DNA microarray ( approximately 2.5 million data points), failed to discriminate between toxicant classes. A major obstacle encountered in these studies was the lack of reproducible gene responses, presumably due to biological variability and technological limitations. Thus multiple replicate observations for the prototypical DNA damaging agent, cisplatin, and the non-steroidal anti-inflammatory drugs (NSAIDs) diflunisal and flufenamic acid were made, and a subset of genes yielding reproducible inductions/repressions was selected for comparison. Many of the "fingerprint genes" identified in these studies were consistent with previous observations reported in the literature (e. g., the well-characterized induction by cisplatin of p53-regulated transcripts such as p21(waf1/cip1) and PCNA [proliferating cell nuclear antigen]). These gene subsets not only discriminated among the three compounds in the learning set but also showed predictive value for the rest of the database ( approximately 100 compounds of various toxic mechanisms). Further refinement of the clustering strategy, using a computer-based optimization algorithm, yielded even better results and demonstrated that genes that ultimately best discriminated between DNA damage and NSAIDs were involved in such diverse processes as DNA repair, xenobiotic metabolism, transcriptional activation, structural maintenance, cell cycle control, signal transduction, and apoptosis. The determination of genes whose responses appropriately group and dissociate anti-inflammatory versus DNA-damaging agents provides an initial paradigm upon which to build for future, higher throughput-based identification of toxic compounds using gene expression patterns alone.

Algorithms↗