27 results on '"Bhatnagar, D."'
Search Results
2. The efficacy of antisense-based construct for inducing resistance against Croton yellow vein mosaic virus in Nicotiana tabacum.
- Author
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Sinha, V., Sarin, N., and Bhatnagar, D.
- Abstract
Begomoviruses have increased pathogenicity because of their adaptation to a wide host range; consequently, these viruses cause a major loss to agroeconomic crops worldwide. In this study, we designed a gene construct representing an antisense coat protein gene. We also analyzed the efficacy of the induced resistance against Croton yellow vein mosaic virus (CrYVMV) affecting papaya in Nicotiana tabacum plants. Positive control plants developed typical leaf curl symptoms, whereas transgenic plants were symptomless. Moreover, the key component (i.e., short interfering RNA) of the antisense pathway was upregulated in transgenic plants. This finding demonstrates the activation of the gene silencing mechanism in transgenic plants. Thus, these results confirm that our construct is functional and effectively induces transient resistance against CrYVMV infections. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
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3. Study of betasatellite molecule from leaf curl disease of sunn hemp ( Crotalaria juncea) in India.
- Author
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Kumar, A., Kumar, J., Khan, Z., Yadav, N., Sinha, V., Bhatnagar, D., and Khan, J.
- Abstract
Leaves of sunn hemp ( Crotalaria juncea) showing geminiviral symptoms were collected from Lucknow, India during rainy season in 2008. DNA template isolated from the symptomatic leaf tissues were subjected to polymerase chain reaction (PCR) using specific primers to amplify coat protein (CP) gene of DNA-A as well as betasatellite DNA associated with the leaf curl disease. CP gene showed 97% sequence identity with that of Cotton leaf curl Burewala virus (CLCuBwV). Further, the betasatellite DNA molecule revealed sequence similarity with previously characterized betasatellite DNA of begomoviruses affecting malvaceous crops from different regions of India and Pakistan. Maximum similarity (>90%) of betasatellite DNA under study was observed with Cotton leaf curl Multan betasatellite (CLCuMB-[Pak: Mul17:08) and other betasatellite DNA from Pakistan thus confirming possible infection of C. juncea with begomovirus. A complementary sense open reading frame (ORF) βC1 is present at nucleotide position 194-550. Sequence comparison of this ORF with other members of begomoviruses further confirmed association of a begomovirus with C. juncea. The betasatellite DNA when expressed under the control of CaMV35S promoter Nicotiana tabacum, showed leaf deformities. Our results demonstrated that a malvaceous betasatellite is adapted by a nonmalvaceous host and causes similar disease symptoms. [ABSTRACT FROM AUTHOR]
- Published
- 2010
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4. Expression of E-Cadherin in breast carcinomas and its association with other biological markers - a prospective study.
- Author
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Chintamani, Rekhi, B., Bansal, A., Bhatnagar, D., and Saxena, S.
- Abstract
Context: E-cadherin (E-CD) is an important cell adhesion molecule in normal epithelial cells and has been shown to be an invasion tumor suppressor gene. Materials and methods: Various clinicopathological parameters like age, family history, tumor stage, histological grade, lymph node status and other biological markers were also analyzed. Present study reveals E-CD expression in 65 cases of breast cancer including 41 (63%) cases of pure infiltrating ductal carcinoma (IDC), 11 (16.9%) of pure infiltrating lobular carcinoma (ILC); another 10 (15.3%) of mixed ductal/lobular type, and remaining 3 (4.6%) miscellaneous types. Results: Negative E-CD expression was noticed more in advancing age groups ( P = 0.01). About 59.2% cases showing negative E-CD expression had family history of breast and/or other cancers. E-CD expression was found significantly higher in cases of pure IDC (55.5%) than in pure ILC cases (18.1%) ( P = 0.04). Eleven (68.7%) of the total 16 high-grade IDC cases, revealed negative expression. Both cases of comedo carcinoma revealed negative expression. Three (30%) out of 10 mixed cases revealed negative expression in both ductal and lobular areas, while in remaining 7 cases, positvity was seen in ductal areas only. Invasive cribriform and medullary carcinoma revealed a stronger expression, while negative staining was observed in sweat gland carcinoma. E-CD re-expression was noticed in lymph node tumor deposits. A direct association of E-CD expression with ER expression and an inverse association with that of p53 were also observed ( P = 0.001), ( P = 0.04). Conclusions: E-CD expression is useful, but limited, in differentiating IDCs from ILCS. Its negative expression correlates with certain poor prognostic parameters reflecting its use as a marker for invasive cancers. It re-expresses at metastatic sites. [ABSTRACT FROM AUTHOR]
- Published
- 2010
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5. Elucidation of veA-dependent genes associated with aflatoxin and sclerotial production in Aspergillus flavus by functional genomics.
- Author
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Cary, J. W., OBrian, G. R., Nielsen, D. M., Nierman, W., Harris-Coward, P., Yu, J., Bhatnagar, D., Cleveland, T. E., Payne, G. A., and Calvo, A. M.
- Subjects
AFLATOXINS ,FUNGI ,GENOMICS ,GENES ,DNA ,STRAINS & stresses (Mechanics) - Abstract
The aflatoxin-producing fungi, Aspergillus flavus and A. parasiticus, form structures called sclerotia that allow for survival under adverse conditions. Deletion of the veA gene in A. flavus and A. parasiticus blocks production of aflatoxin as well as sclerotial formation. We used microarray technology to identify genes differentially expressed in wild-type veA and veA mutant strains that could be involved in aflatoxin production and sclerotial development in A. flavus. The DNA microarray analysis revealed 684 genes whose expression changed significantly over time; 136 of these were differentially expressed between the two strains including 27 genes that demonstrated a significant difference in expression both between strains and over time. A group of 115 genes showed greater expression in the wild-type than in the veA mutant strain. We identified a subgroup of veA-dependent genes that exhibited time-dependent expression profiles similar to those of known aflatoxin biosynthetic genes or that were candidates for involvement in sclerotial production in the wild type. [ABSTRACT FROM AUTHOR]
- Published
- 2007
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6. Amino acid supplementation reveals differential regulation of aflatoxin biosynthesis in Aspergillus flavus NRRL 3357 and Aspergillus parasiticus SRRC 143.
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Wilkinson, J. R., Yu, J., Bland, J. M., Nierman, W. C., Bhatnagar, D., and Cleveland, T. E.
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AFLATOXINS ,METABOLITES ,ASPERGILLUS flavus ,SUCROSE ,BIOSYNTHESIS ,GENETICS - Abstract
Aflatoxins are toxic and carcinogenic secondary metabolites produced by the fungi Aspergillus flavus and Aspergillus parasiticus. To better understand the molecular mechanisms that regulate aflatoxin production, the biosynthesis of the toxin in A. flavus and A. parasticus grown in yeast extract sucrose media supplemented with 50 mM tryptophan (Trp) were examined. Aspergillus flavus grown in the presence of 50 mM tryptophan was found to have significantly reduced aflatoxin B
1 and B2 biosynthesis, while A. parasiticus cultures had significantly increased B1 and G1 biosynthesis. Microarray analysis of RNA extracted from fungi grown under these conditions revealed 77 genes that are expressed significantly different between A. flavus and A. parasiticus, including the aflatoxin biosynthetic genes aflD (nor-1), aflE (norA), and aflO (omtB). It is clear that the regulatory mechanisms of aflatoxin biosynthesis in response to Trp in A. flavus and A. parasiticus are different. These candidate genes may serve as regulatory factors of aflatoxin biosynthesis. [ABSTRACT FROM AUTHOR]- Published
- 2007
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7. CYP17 gene polymorphism and its association with high-risk north Indian breast cancer patients.
- Author
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Chakraborty, Anurupa, Murthy, N. S., Chintamani, Chintamani, Bhatnagar, D., Mohil, R. S., Sharma, P. C., and Saxena, Sunita
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GENETIC polymorphisms ,DISEASE risk factors ,BREAST cancer ,CANCER patients ,CANCER in women - Abstract
A single T > C change at the 5′ promoter region of the CYP17 gene is reported to be associated with increased risk of breast cancer. This study evaluates the influence of genetic polymorphism of CYP17 on breast cancer susceptibility. Two hundred and forty-two patients with histopathologically confirmed breast cancer and 212 age-matched controls were included in the present study. Information relating to age at onset of the disease, family history and estrogen receptor status was elicited. Investigation for CYP17 polymorphism was carried out in 106 early onset, 80 late onset and 56 familial cases. The frequencies of two CYP17 alleles were also analyzed in 116 (47.9%) cases with known estrogen receptor (ER) status confirmed immunohistochemically. A polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) method was used to detect the polymorphism, and the genotypes identified were assigned as homozygous wild type (A1A1), heterozygous variant (A1A2), and homozygous variant (A2A2). Associations between the various genotypes in patients and controls were investigated with Fisher’s exact test. All the tests were two tailed. The results showed that the frequency of heterozygous and homozygous CYP17 genotype was higher in early onset breast cancer patients (94.3%) than in controls (80.3%), and the difference was significant ( P = 0.001). A highly statistically significant increased risk in carriers of homozygous A2 allele was found in young patients ( P ≤ 0.001) in comparison with patients having late onset condition ( P = 0.260). However, no significant association between the genotype and breast cancer risk was observed among women with strong family history. Further, data had showed that patients (80.6%) with at least one A2 allele tended to exhibit ER-independent cell proliferation, although statistical significance could not be established ( P = 0.160). The present findings suggest that CYP17 A2 allele gene polymorphism might play a significant role in breast cancer development in young Indian women. [ABSTRACT FROM AUTHOR]
- Published
- 2007
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8. Regulatory elements in aflatoxin biosynthesis.
- Author
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Cary, J., Ehrlich, K., Kale, S., Calvo, A., Bhatnagar, D., and Cleveland, T.
- Abstract
This review provides a synopsis of factors involved in the regulation of aflatoxin in Aspergillus species at the molecular level. Much of the knowledge available today on the regulation of secondary metabolite production in fungi has been gleaned from studies of the aflatoxin gene cluster in A. flavus and A. parasiticus and the sterigmatocystin gene cluster in A. nidulans. Regulation of these two gene clusters is under the control of both pathway-specific transcription factors such as AflR and AflJ and global or broad-domain transcription factors such as AreA and PacC. Study of secondary metabolite (sec-) mutants in A. parasiticus first identified an association between mycotoxin production and fungal development. This linkage has been extended at the molecular level by the characterization of a G-protein/cAMP/Protein kinase A signaling pathway that regulates sporulation via the transcription factor BrlA and aflatoxin/sterigmatocystin production via AflR. Another global regulator of mycotoxin production, VeA, mediates a developmental light-response in A. nidulans and A. flavus. Though not similar to any known fungal transcriptional regulators, VeA controls aflatoxin/sterigmatocystin production via transcriptional control of AflR and it also regulates development of sexual structures such as cleistothecia in A. nidulans and sclerotia in A. flavus. [ABSTRACT FROM AUTHOR]
- Published
- 2006
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9. Gene targets for fungal and mycotoxin control.
- Author
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Kim, J., Campbell, B., Molyneux, R., Mahoney, N., Chan, K., Yu, J., Wilkinson, J., Cary, J., Bhatnagar, D., and Cleveland, T.
- Abstract
It was initially shown that gallic acid, from hydrolysable tannins in the pelliele of walnut kernels, dramatically inhibits biosynthesis of aflatoxin by Aspergillus flavus. The mechanism of this inhibition was found to take place upstream from the gene cluster, including the regulatory gene, aflR, involved in aflatoxin biosynthesis. Additional research using other antioxidant phenolics showed similar antiaflatoxigenic activity to gallic acid. Treatment of A. flavus with tert-butyl hydroperoxide resulted in an almost doubling of aflatoxin biosynthesis compared to untreated samples. Thus, antioxidative response systems are potentially useful molecular targets for control of A. flavus. A high throughput screening system was developed using yeast, Saccharomyces cerevisiae, as a model fungus. This screening provided an avenue to quickly identify fungal genes that were vulnerable to treatment by phenolic compounds. The assay also provided a means to quickly assess effects of combinations of phenolics and certain fungicides affecting mitochondrial respiration. For example, the S. cerevisiae sod2† mutant was highly sensitive to treatment by certain phenolics and strobilurins/antimycin A, fungicides which inhibit complex III of the mitochondrial respiratory chain. Verification of stress to this system in the target fungus, A. flavus, was shown through complementation analysis, wherein the mitochondrial superoxide dismutase (Mn-SOD) gene ( sodA) of A. flavus in the ortholog mutant, sod2†, of S. cerevisiae, relieved phenolic-induced stress. Mitochondrial antioxidative stress systems play an important role in fungal response to antifungals. Combined treatment of fungi with phenolics and inhibitors of mitochondrial respiration can effectively suppress growth of A. flavus in a synergistic fashion. [ABSTRACT FROM AUTHOR]
- Published
- 2006
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10. Aspergillus flavus expressed sequence tags and microarray as tools in understanding aflatoxin biosynthesis.
- Author
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Yu, J., Cleveland, T., Wilkinson, J., Campbell, B., Kim, J., Kim, H., Bhatnagar, D., Payne, G., and Nierman, W.
- Abstract
Aflatoxins are the most toxic and carcinogenic naturally occurring mycotoxins. They are produced primarily by Aspergillus flavus and A. parasiticus. In order to better understand the molecular mechanisms that control aflatoxin production, identification of genes using A. flavus expressed sequence tags (ESTs) and microarrays is currently being performed. Sequencing and annotation of A. flavus ESTs from a normalized A. flavus cDNA library identified 7,218 unique EST sequences. Genes that are putatively involved in aflatoxin biosynthesis, regulation and signal transduction, fungal virulence or pathogenicity, stress response or antioxidation, and fungal development were identified from these ESTs. Microarrays containing over 5,000 unique A. flavus gene amplicons were constructed at The Institute for Genomic Research. Gene expression profiling under aflatoxin-producing and non-producing conditions using this microarray has identified hundreds of genes that are potentially involved in aflatoxin production. Further investigations on the functions of these genes by gene knockout experiments are underway. This research is expected to provide information for developing new strategies for controlling aflatoxin contamination of agricultural commodities. [ABSTRACT FROM AUTHOR]
- Published
- 2006
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11. Marked differences in the IGF system that are associated with migration in comparable populations of Gujaratis living in Sandwell, UK, and Gujarat, India.
- Author
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Heald, A. H., Anderson, S. G., Vyas, A., Siddals, K., Patel, J., Yates, A. P., Bhatnagar, D., Prabhakaran, D., Hughes, E., Rudenski, A., Durrington, P., Gibson, J. M., and Cruickshank, J. K.
- Subjects
LIFESTYLES ,EMIGRATION & immigration ,CARDIOVASCULAR diseases ,SOMATOMEDIN ,GLUCOSE - Abstract
Aims/hypotheses: We previously reported independent links between the IGF system and the development of impaired glucose tolerance and cardiovascular risk. This study tests the hypothesis that the lifestyle change which accompanies population migration, with attendant increases in cardiovascular risk, is reflected by changes in the IGF system. Materials and methods: We compared a specific Gujarati community in Sandwell, UK (n=205), with people still resident in the same villages of origin near Navsari, India (n=246). We performed anthropometry and measured fasting plasma insulin, IGF-I, insulin-like growth factor binding protein (IGFBP)-1 and IGFBP-3. Results: Daily calorie intake, BMI and WHR were significantly higher in UK Gujaratis than in Indian Gujaratis. IGFBP-1 was significantly lower in UK migrants (mean 29.5 [95% CI 25.9–33.0] vs 56.5 [50.6–62.5]μg/l; F=48.4, p«0.001). Conversely, fasting insulin, IGFBP-3 and IGF-I were all higher in UK Gujaratis (mean IGF-I 145.9 [138.1–153.6]ng/ml in UK Gujaratis and 100.9 [94.6–107.3] ng/ml in Navsari Gujaratis; F=76.6, p«0.001). These differences were still apparent when adjustment was made for BMI by location for IGF-I (F=57.4, p«0.001) and IGFBP-3 (F=5.7, p=0.02), but were no longer apparent for IGFBP-1 and insulin. At the population level, the decrease in IGFBP-1 for a given increase in insulin was significantly smaller in UK Gujaratis, suggesting greater hepatic insulin resistance in this group. Conclusions/interpretation: Environmental factors have profound effects on circulating IGF system components and on the relationship between IGFBP-1, IGF-I and related metabolic variables. This may have long-term implications for the development of worsening glucose tolerance and cardiovascular disease. [ABSTRACT FROM AUTHOR]
- Published
- 2005
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12. Molecular genetic analysis and regulation of aflatoxin biosynthesis.
- Author
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Bhatnagar, D., Ehrlich, K. C., and Cleveland, T. E.
- Subjects
AFLATOXINS ,ASPERGILLUS ,GENES ,DNA-binding proteins ,MONOOXYGENASES ,FUNGI - Abstract
Aflatoxins, produced by some Aspergillus species, are toxic and extremely carcinogenic furanocoumarins. Recent investigations of the molecular mechanism of AFB biosynthesis showed that the genes required for biosynthesis are in a 70 kb gene cluster. They encode a DNA-binding protein functioning in aflatoxin pathway gene regulation, and other enzymes such as cytochrome P450-type monooxygenases, dehydrogenases, methyltransferases, and polyketide and fatty acid synthases. Information gained from these studies has led to a better understanding of aflatoxin biosynthesis by these fungi. The characterization of genes involved in aflatoxin formation affords the opportunity to examine the mechanism of molecular regulation of the aflatoxin biosynthetic pathway, particularly during the interaction between aflatoxin-producing fungi and plants. [ABSTRACT FROM AUTHOR]
- Published
- 2003
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13. Should Pediatric Patients with Hyperlipidemia Receive Drug Therapy?
- Author
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Bhatnagar, D.
- Subjects
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HYPERLIPIDEMIA in children , *JUVENILE diseases , *CORONARY disease - Abstract
Hyperlipidemia is now established as a major risk factor for causation of coronary heart disease (CHD) in adults; however, there is much debate on the level of coronary risk at which lipid-lowering drugs should be used. These issues of possible harm or lack of benefit from long-term use of lipid-lowering therapy, and cost effectiveness, are also pertinent in the pediatric setting. Evidence from several countries indicates that children have an increasing prevalence of obesity, hyperlipidemia and type 2 diabetes mellitus. Children who have high serum lipids ‘track’ these increased levels into adulthood. In some countries there is a trend to screen children for hypercholesterolemia. Family history itself is a poor discriminator in determining which children need to be screened and treated. Estimation of apolipoprotein B and/or apolipoprotein E genotype can improve prediction. Measuring high density lipoprotein cholesterol also helps, but obesity appears to be the best marker for screening children at high risk. These considerations should not cloud the need for case finding and treatment of children with genetic disorders. Low fat diets have been shown to be well tolerated and effective in children; however, there are no major long-term studies demonstrating harm or benefit in those on lipid-lowering drugs. Nevertheless, concerns regarding the psychological effect and the theoretical metabolic effects of long-term lipid lowering remain. Lipid-lowering drugs should be generally restricted to children with genetic disorders of lipid metabolism. Children with diabetes mellitus, hypertension or nonlipid-related inherited disorders leading to premature CHD in adults should be treated with diet, and with lipid-lowering drugs when they reach adulthood. Children with secondary hyperlipidemia should be assessed individually. A number of drugs and nutriceuticals are available for use in children, but only a few drugs are licensed for use in children. [ABSTRACT FROM AUTHOR]
- Published
- 2002
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14. Growth inhibition of a Fusarium verticillioides GUS strain in corn kernels of aflatoxin-resistant genotypes.
- Author
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Brown, R. L., Cleveland, T. E., Woloshuk, C. P., Payne, G. A., and Bhatnagar, D.
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CORN ,ASPERGILLUS flavus ,ASPERGILLOSIS ,AFLATOXINS ,FUSARIUM ,GLUCURONIDASE ,GENES - Abstract
Two corn genotypes, GT-MAS:gk and MI82, resistant to Aspergillus flavus infection/aflatoxin contamination, were tested for their ability to limit growth of Fusarium verticillioides. An F. verticillioides strain was transformed with a β-glucuronidase (GUS) reporter gene (uidA) construct to facilitate fungal growth quantification and then inoculated onto endosperm-wounded and non-wounded kernels of the above-corn lines. To serve as a control, an A. flavus strain containing the same reporter gene construct was inoculated onto non-wounded kernels of GT-MAS:gk. Results showed that, as in a previous study, non-wounded GT-MAS:gk kernels supported less growth (six- to ten-fold) of A. flavus than did kernels of a susceptible control. Also, non-wounded kernels of GT-MAS:gk and MI82 supported less growth (two- to four-fold) of F. verticillioides than did susceptible kernels. Wounding, however, increased F. verticillioides infection of MI82, but not that of GT-MAS:gk. This is in contrast to a previous study of A. flavus, where wounding increased infection of GT-MAS:gk rather than MI82 kernels. Further study is needed to explain genotypic variation in the kernel response to A. flavus and F. verticillioides kernel infections. Also, the potential for aflatoxin-resistant corn lines to likewise inhibit growth of F. verticillioides needs to be confirmed in the field. [ABSTRACT FROM AUTHOR]
- Published
- 2001
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15. Pre-termination in aflR of Aspergillus sojae inhibits aflatoxin biosynthesis.
- Author
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Matsushima, K., Chang, P.-K., Yu, J., Abe, K., Bhatnagar, D., and Cleveland, T. E.
- Subjects
ASPERGILLUS ,MONILIACEAE ,ASPERGILLUS flavus ,AFLATOXINS ,MYCOTOXINS ,BIOSYNTHESIS - Abstract
The aflR gene product is the main transcriptional regulator of aflatoxin biosynthesis in Aspergillus parasiticus and Aspergillus flavus. Although A. sojae strains do not produce aflatoxins, they do have an aflR homologue. When compared with the aflR of A. parasiticus, the A. sojae gene contains two mutations: an HAHA motif and a premature stop codon. To investigate the functionality of the A. sojae aflR gene product, we used a GAL4 one-hybrid system in yeast. The transcription-activating activity of AflR from A. sojae was 15% of that from A. parasiticus. The introduction of an additional aflR from A. sojae into an A. parasiticus strain did not affect aflatoxin productivity. A hybrid aflR comprising the amino-terminal region of A. sojae aflR and the carboxy-terminal region of A. parasiticus aflR suppressed the effect associated with pre-termination of the A. sojae AflR. We conclude that the premature stop codon of the A. sojae aflR is the key to its functionality and leads to prevention of aflatoxin biosynthesis through loss of the transcription of aflatoxin biosynthesis-related genes. [ABSTRACT FROM AUTHOR]
- Published
- 2001
- Full Text
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16. Generation of aflR disruption mutants of Aspergillus parasiticus.
- Author
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Cary, J. W., Ehrlich, K. C., Wright, M., Chang, P.-K., and Bhatnagar, D.
- Subjects
ASPERGILLUS ,DNA ,PROTEINS ,AFLATOXINS ,BIOSYNTHESIS - Abstract
The aflR gene of Aspergillus parasiticus and A. flavus encodes a binuclear zinc-finger, DNA-binding protein, AflR, responsible for activating the transcription of all known aflatoxin biosynthetic genes including itself. Studies to determine how environmental and nutritional factors affect aflR expression and hence aflatoxin production in A. parasiticus have been difficult to perform due to the lack of aflR“knockout” mutants. Transformation of an O-methylsterigmatocystin (OMST)-accumulating strain of A. parasiticus with an aflR-niaD gene disruption vector resulted in clones harboring a recombinationally inactivated aflR gene which no longer produced OMST or aflR transcript. By transformation of this aflR disruptant strain with constructs containing mutated versions of the aflR promoter, we identified three cis-acting sites that were necessary for aflR function: an AflR-binding site, a PacC-binding site, and a G + A-rich site near the transcription start site of aflR. [ABSTRACT FROM AUTHOR]
- Published
- 2000
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17. Genes encoding cytochrome P450 and monooxygenase enzymes define one end of the aflatoxin pathway gene cluster in Aspergillus parasiticus.
- Author
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Yu, J., Chang, P.-K., Bhatnagar, D., and Cleveland, T. E.
- Subjects
GENES ,CYTOCHROMES ,HEMOPROTEINS ,ENZYMES ,AFLATOXINS ,ASPERGILLUS - Abstract
The identification of overlapping cosmids resulted in the discovery of the aflatoxin biosynthetic pathway gene cluster in Aspergillus flavus and A. parasiticus. This finding led to the cloning and characterization of one regulatory and 16 structural genes involved in aflatoxin biosynthesis, including the most recent report on the gene, ordA, which has been identified to be involved in the formation of four aflatoxins (B
1 , B2 , G1 and G2 ). However, these genes do not account for all the identified chemical/biochemical steps in aflatoxin synthesis and efforts are underway to identify the genes controlling the other steps. We are also attempting to define the outer boundaries of the aflatoxin pathway gene cluster in the Aspergillus genome. For this goal, we extended sequencing in both directions from the existing (60 kb) aflatoxin pathway gene cluster, beyond the pksA gene at one end and the omtA gene at the other. Within the 25-kb genomic DNA sequence determined at the omtA end of the cluster, several new gene sequences were identified. The recently reported genes, vbs and ordA, were found within this 25-kb region. Two additional genes were also found in this region, a cytochrome P450 monooxygenase encoding gene, tentatively named cypX, and a monooxygenase encoding gene, tentatively named moxY, and these are also reported in this study. The sequence beyond these genes showed a 5-kb non-coding region of DNA followed by the presence of a cluster of genes probably involved in sugar metabolism. Northern blot analysis and reverse transcriptase-polymerase chain reaction (RT-PCR) studies demonstrated that the genes, cypX and moxY, are expressed concurrently with genes involved in aflatoxin biosynthesis. Therefore, the two putative aflatoxin pathway genes cypX and moxY followed by a 5-kb non-coding region of DNA define one end of the boundary of the aflatoxin pathway gene cluster in A. parasiticus. [ABSTRACT FROM AUTHOR]- Published
- 2000
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18. Isolation and characterization of experimentally induced, aflatoxin biosynthetic pathway deletion mutants of Aspergillus parasiticus.
- Author
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Cary, J. W., Barnaby, N., Ehrlich, K. C., and Bhatnagar, D.
- Abstract
A plasmid vector (pDEL2) was engineered for the purpose of introducing a deletion within the aflatoxin (AF) biosynthetic gene cluster of Aspergillus parasiticus. The vector was constructed by PCR amplification of a region of the AF gene cluster from an A. parasiticus isolate that had undergone an aberrant recombinational event during transformation with a norA-niaD gene disruption vector. This recombinational event resulted in the deletion of an approximately 6-kb region of the AF gene cluster and accumulation of the AF precursor averantin (AVN). Northern hybridization analysis confirmed that the deletion event resulted in no detectable transcription of the norA gene or the AF biosynthetic genes, avnA, verA, and ver-1. Transformation of A. parasiticus RHN1 with pDEL2 resulted in 16% of the transformants accumulating AVN. Southern hybridization analysis of randomly selected AVN-accumulating transformants indicated that all had undergone a double-crossover homologous, recombinational event resulting in the 6-kb norA to avnA deletion within the AF gene cluster. Aflatoxin precursor feeding studies performed on one of the AVN-accumulating, RHN1(pDEL2) transformants confirmed that the enzyme activities associated with the deleted genes were no longer present. [ABSTRACT FROM AUTHOR]
- Published
- 1999
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19. Synthesis of sterigmatocystin on a chemically defined medium by species of Aspergillus and Chaetomium.
- Author
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Barnes, S., Dola, T., Bennett, J., and Bhatnagar, D.
- Abstract
Sterigmatocystin (ST) is a secondary metabolite and a principal mycotoxin known to be produced by over 30 species of filamentous fungi. It is also one of the late intermediates in aflatoxin biosynthesis. We have tested the ability of 7 species of Aspergillus, including 4 strains of A. versicolor, one species of Bipolaris, and two species of Chaetomium, to produce ST on a sucrose-salts-phenylalanine defined medium as well as on three complex substrates. Highest ST production in our survey was by a strain of A. versicolor grown on wheat, whereas, the highest ST production on defined medium was by C. cellulolyticum. To our knowledge, this is the first report of ST production by C. cellulolyticum on any substrate. In precursor feeding studies, resting cultures of wild type A. nidulans and A. versicolor were unable to biotransform O-methylsterigmatocystin (OMST), the last known intermediate in aflatoxin biosynthesis. These results suggest that ST is the end product of polyketide metabolism in the strains tested. [ABSTRACT FROM AUTHOR]
- Published
- 1994
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20. A study of physical growth in breast-fed and bottle-fed male infants.
- Author
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Sidhu, L., Grewal, R., and Bhatnagar, D.
- Abstract
Data for ten anthropometric variables has been collected on a cross sectional sample of 54 male infants varying in age from 3 to 15 months. Of these, 29 infants were completely dependent on breast milk and 25 were those who took only bottle milk. All the anthropometric parameters studied indicated that breastfed infants were better in physical growth upto 7 months of age as compared to the bottle-fed infants of the same age group. After that bottlefed infants show better physical growth than breast-fed infants. It is recommended that in breast-fed children weaning should not be delayed beyond 7th mouth. [ABSTRACT FROM AUTHOR]
- Published
- 1981
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21. Hybridization of genes involved in aflatoxin biosynthesis to DNA of aflatoxigenic and non-aflatoxigenic aspergilli.
- Author
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Klich, M., Yu, J., Chang, P.-K., Mullaney, E., Bhatnagar, D., and Cleveland, T.
- Abstract
Southern blots of DNA from a number of aspergilli belonging to Aspergillus section Flavi, including aflatoxin-producing and non-aflatoxigenic isolates of A. flavus and A. parasiticus, were probed with the aflatoxin pathway genes aflR and omt-1. DNA of all A. flavus, A. parasiticus and A. sojae isolates examined hybridized with both genes. None of the A. oryzae isolates examined hybridized to the aflR probe and one of the three did not hybridize to the omt-1 probe. None of the A. tamarii isolates examined hybridized to either gene. Our results suggest that some isolates in this section do not produce aflatoxin because they lack at least one of the genes necessary for biosynthesis, and that non-producing A. flavus, A. parasiticus and A. sojae strains either lack a gene we did not examine or have genes that are not being expressed. [ABSTRACT FROM AUTHOR]
- Published
- 1995
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22. Characterization of the Aspergillus parasiticus niaD and niiA gene cluster.
- Author
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Chang, Perng-Kuang, Ehrlich, Kenneth C., Linz, John E., Bhatnagar, D., Cleveland, Thomas E., and Bennett, Joan W.
- Abstract
The nitrate reductase gene ( niaD) and nitrite reductase gene ( niiA) of Aspergillus parasiticus are clustered and are divergently transcribed from a 1.6-kb intergenic region ( niaD-niiA). The deduced aminoacid sequence of the A. parasiticus nitrate reductase demonstrated a high degree of homology to those of other Aspergillus species, as well as to Leptosphaeria maculans, Fusarium oxysporum, Gibberella fujikuroi and Neurospora crassa, particularly in the cofactor-binding domains for molybdenum, heme and FAD. A portion of the deduced nitrite reductase sequence was homologous to those of A. nidulans and N. crassa. The nucleotide sequences in niaD-niiA of A. parasiticus and of A. oryzae were 95% identical, indicating that these two species are closely related. Several GATA motifs, the recognition sites for the N. crassa positive-acting global regulatory protein NIT2 in nitrogen metabolism, were found in A. parasiticus niaD-niiA. Two copies of the palindrome TCCGCGGA and other partial palindromic sequences similar to the target sites for the pathway specific regulatory proteins, N. crassa NIT4 and A. nidulans NirA, in nitrate assimilation, were also identified. A recombinant protein containing the A. nidulans AreA (the NIT2 equivalent) zinc finger and an adjacent basic region was able to bind to segments of niaD-niiA encompassing the GATA motifs. These results suggest that the catalytic and regulatory mechanisms of nitrate assimilation are well conserved in Aspergillus. [ABSTRACT FROM AUTHOR]
- Published
- 1996
- Full Text
- View/download PDF
23. Lipid peroxidative damage on cadmium exposure and alterations in antioxidantsystem in rat erythrocytes: A study with relation to time.
- Author
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Sarkar, S., Yadav, P., and Bhatnagar, D.
- Abstract
Cadmium induced lipid peroxidation (LPO) and the activity of antioxidantenzymes after the administration of a single dose of CdCl 2 (0.4 mg kg body wt, ip) was studied in rat erythrocytes.Cd intoxication increased erythrocyte LPO along with a decrease insuperoxide dismutase (SOD) up to three days of Cd treatment. Thedecrease in erythrocyte catalase (CAT) activity was marked within9 h of Cd intoxication. After three days of Cd treatment, LPOdecreased towards normal, along with an increase in erythrocyteSOC and CAT activity. Blood glutathione (GSH) decreased significantlywithin 24 h of Cd treatment, followed by an increase towards normal.Erythrocyte glutathione S-transferase (GST) activity increased up to10 days of Cd intoxication, probably in an attempt to reduce Cd toxicity.Serum glutamate pyruvate transaminase (SGPT), serum alkaline phosphatase(SALP) and serum bilirubin increased up to 10 days of Cd intoxication.Blood urea increased significantly up to three days, followed by a decreasetowards normal. The results show that Cd induced LPO was associated with adecrease in antioxidant enzymes and GSH in erythrocytes; as these antioxidantsincrease in erythrocytes with recovery from Cd intoxication, the Cd inducedLPO reversed towards normal. The increase in the SGPT, SALP and serum bilirubincorrelated with LPO. The results suggest that Cd intoxication induces oxidativestress and alters the antioxidant system, resulting in oxidative damage torat erythrocytes. © Rapid Science 1998 [ABSTRACT FROM AUTHOR]
- Published
- 1998
- Full Text
- View/download PDF
24. Effect of N-Nitrosodiethylamine on Lipid Peroxidation and Antioxidant Enzymes in Rat Liver Mitochondria: Protective Role of Antioxidants.
- Author
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Bansal, A. K., Bhatnagar, D., and Soni, G. L.
- Subjects
DIMETHYLNITROSAMINE ,ANTIOXIDANTS ,PEROXIDATION ,NITROSOAMINES ,ANIMAL products ,MITOCHONDRIAL DNA ,MITOCHONDRIA ,NITROSO compounds ,MEAT - Abstract
The article examines the effect of N-nitrosodiethylamine (NDEA) on lipid peroxidation (LPO) and the antioxidant defense enzymes in an isolated rat liver mitochondria. It is said that the NDEA have been suggested to cause oxidative stress and cellular injury as it is being implicated in the aetiology of some human cancers. The study was based on the nitrosamines, the N-nitroso compounds in rat and some in food such as meat, dairy products and alcoholic beverages. After several experimentation, results indicates an increase in the LPO when it was exposed to different concentrations of NDEA. Several charts depicting the experiment results are presented for further information.
- Published
- 1997
- Full Text
- View/download PDF
25. Lipid peroxidation and activities of antioxygenic enzymes in vitro in mercuric chloride treated human erythrocytes.
- Author
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Erythrocytes, Human, Bansal, A., Bhatnagar, D., and Bhardwaj, R.
- Subjects
MERCURIC chloride ,LIPID peroxidation (Biology) ,ERYTHROCYTES ,ENZYMES ,GLUTATHIONE - Abstract
The article discusses a study which examined the in vitro toxicity of mercuric chloride on human erythrocytes, in particular its effect on lipid peroxidation (LPO) and activities of antioxygenic enzymes. Blood was collected from 30 healthy male donors and red blood cells were harvested The packed cells were pretreated with mercuric chloride at various concentrations and different intervals. The study found that mercuric chloride caused an increased in LPO and decline in glutathione content.
- Published
- 1992
- Full Text
- View/download PDF
26. Xanthogranulomatous pyelonephritis.
- Author
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Gera, Neeru, Bhatia, Arati, Bhatnagar, Dinesh, Gupta, Sanjay, Gera, N, Bhatia, A, Bhatnagar, D, and Gupta, S
- Published
- 1993
- Full Text
- View/download PDF
27. Effect of Exposure to Sun and Exercise on Heat Tolerance Coefficient in Murrah Buffalo Calves.
- Author
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BHATNAGAR, D. S. and CHAUDHARY, N. C.
- Published
- 1961
- Full Text
- View/download PDF
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