Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.6.1.44 (AGT)
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Using three antibodies (JG8, CM-1 and 1081) directed to the p53 protein, strong positivity was found in 16/47 (34.0%) of mucosal squamous cell carcinomas of the head and neck and in two squamous carcinoma cell lines (LICR-LON- HN5 and HN6Rr). The presence of the mutant p53 was confirmed in the cell lines as substitutions in exon 7 (codon 238, TGT greater than AGT) and exon 5 (codon 152, CCG greater than CTG) respectively. Positive staining was seen only in the undifferentiated cells and progressively lost as the cells keratinized, both in the tumour specimens and in the cell lines. Similar results were seen in areas of dysplasia, well removed from the site of the primary tumour. Staining of epidermal lesions showed positivity in 2/12 (16.6%) cases of Bowen's disease, 0/12 (0.0%) cases of solar keratosis, 0/10 (0.0%) basal cell carcinomas and in 3/20 (15.0%) squamous cell carcinomas. These results are discussed in relation to the multifocal origin of squamous cell carcinomas, the role of p53 mutations in squamous cell carcinomas from different sites and the significance of the 'basal' distribution of p53 as a normal growth regulator. The possible significance of the distribution of p53 in squamous epithelium as it relates to papilloma virus infection is also considered.
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PMID:Expression of p53 in premalignant and malignant squamous epithelium. 171 23

To examine the significance of mutation of the p53 tumour suppressor gene in the development of human hepatocellular carcinoma in a high-prevalence area for hepatitis B viral infection but a low-exposure area for aflatoxin B1, the spectrum of p53 gene mutations was examined in 21 tumour samples from Hong Kong Chinese patients, all of whom were HBsAg positive. DNA sequencing covering exons 5 to 9 of the p53 gene and Hae III restriction enzyme digestion for preliminary assessment of mutation at codon 249 were performed. Immunohistochemical staining with anti-p53 monoclonal antibodies was done on both tumour and nontumour liver tissues. Six tumours (28.6%) showed a p53 mutation and all were point mutations. Of the six point mutations, two (9.5%) were at codon 249 and both were G to T transversions (AGG-->ATG and AGG-->AGT transversions). The remaining point mutations were transversions scattered at codon 172 (exon 5), 214 (exon 6), 273 (exon 8) and 330 (exon 9). Mutated p53 protein was detected in five of these six cases with demonstrable point mutations by DNA sequencing, in contrast to none detected in all of the 15 cases without demonstrable point mutations. The presence of p53 mutations, including those at codon 249, did not show a significant association with tumour size, sex, age, tumour invasiveness in terms of liver invasion, microsatellites and venous permeation, cirrhosis and encapsulation, but tumours with low cellular differentiation tended to have a higher incidence (71%) of point mutations than those with high cellular differentiation (8%). In conclusion, both the overall p53 mutation rate and that a codon 249 in HCC in Hong Kong Chinese are lower than those reported in tumours from China and sub-Saharan Africa. The low mutation rate at codon 249 is compatible with a low aflatoxin exposure. A special type of p53 mutation has not been found to be associated with hepatitis B viral infection. Mutations of p53 gene tends to occur in tumours with low cellular differentiation, suggesting a late occurrence in the event of tumour progression.
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PMID:p53 gene mutation spectrum in hepatocellular carcinomas in Hong Kong Chinese. 810 45

The baculovirus Bombyx mori nucleopolyhedrovirus (BmNPV) contains five related open reading frames (ORFs). Recent sequence analyses of several other baculovirus genomes reveal that these ORFs belong to a unique multigene family called the baculovirus repeated ORFs (bro) family. Here we have characterized these five genes from BmNPV at the transcriptional and translational levels. Reverse transcription-PCR and primer extension analyses indicated that transcription of all bro genes occurs by 2 to 4 h postinfection (p.i.) and reaches maximal levels between at 8 and 12 h p.i. Transcription of all genes is initiated between 50 and 70 nucleotides upstream of the start codon, at a characteristic C(T)AGT motif. Expression of a cat reporter gene under the control of each bro promoter provides evidence that a viral factor(s) is required for the transcription of all bro genes. Immunoblot analysis indicated that a population of BRO proteins is produced vigorously between at 8 and 14 h p.i. Immunohistochemical analysis by confocal microscopy showed that BRO proteins are localized in both the nucleus and the cytoplasm at 8 h p.i. Four BmNPV mutants, in which the bro-a, bro-b, bro-c, and bro-e genes were individually inactivated, were successfully isolated. However, exhaustive efforts failed to isolate a bro-d-deficient mutant. Similarly, it was not possible to isolate a double-deletion bro-a bro-c mutant. The bro-d gene may play an irreplaceable functional role(s) during viral infection, while bro-a and bro-c may functionally complement each other.
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PMID:Characterization of baculovirus repeated open reading frames (bro) in Bombyx mori nucleopolyhedrovirus. 1055 52

An attempt was made to determine whether amino acid variation at position 631 in the chicken Mx protein definitely influences antiviral specificity, using an artificial mutation technique by which a single amino acid was reciprocally substituted between Ser (AGT) and Asn (AAT) at position 631 of the negative and positive chicken Mx, respectively. Using permanently transfected 3T3 cell lines, the antiviral potential of chicken Mx against vesicular stomatitis virus infection was analysed. The results indicated that the phenotype of antiviral activity depends on the amino acid difference at position 631; that is, the genotype coding Asn at position 631 corresponds to the positive antiviral phenotype, and the genotype coding Ser corresponds to the negative phenotype. The present study has confirmed that the antiviral specificity of chicken Mx protein is determined by an amino acid substitution at the carboxy terminus.
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PMID:Native antiviral specificity of chicken Mx protein depends on amino acid variation at position 631. 1502 71

In recent years, wine grape (Vitis vinifera) acreage in Idaho has expanded because of favorable climatic conditions for premium wine production. Nearly 95% of the 491.7 ha (1,215 acres) of wine grapes are in the Snake River Valley with Canyon County accounting for 81% of the vines. Previous studies have shown that grapevine leafroll disease (GLD) is the most widespread and economically significant virus disease in wine grapes in Washington and Oregon (1,2). However, little is known about the incidence and economic impact of GLD on wine grapes in Idaho. During the 2008 growing season, leaf samples were collected from approximately 25 individual grapevines of red-berried cultivars (Cabernet Sauvignon, Merlot, Syrah, and Petit Syrah) showing GLD symptoms and white-berried (Chardonnay) cultivars with suspected GLD symptoms growing in 10 geographically separate vineyards in Canyon County. An additional five samples were collected from a Lemberger block in Elmore County. Petiole extracts from these samples were tested by single-tube reverse transcription (RT)-PCR with primers LC 1 (5'-CGC TAG GGC TGT GGA AGT ATT-3') and LC 2 (5'-GTT GTC CCG GGT ACC AGA TAT-3') specific for the heat shock protein 70 homologue (HSP-70 gene) of Grapevine leafroll-associated virus-3 (GLRaV-3) (3). All samples, except the Petit Syrah, produced a single band of the expected size of 546 bp. ELISA with GLRaV-3-specific antibodies (BIOREBA AG, Reinach, Switzerland) confirmed the presence of the virus in samples that were positive in RT-PCR. GLRaV-3-specific amplicons were cloned in pCR2.1 plasmid (Invitrogen Corp., Carlsbad, CA) and 2 to 3 independent clones per isolate were sequenced in both orientations. A pairwise comparison of 22 sequences, six from Chardonnay (GenBank Accessions GQ344810, GQ344811, GQ344823, GQ344824, GQ344825, and GQ344826), five from Cabernet Sauvignon (GQ344807, GQ344808, GQ344809, GQ344827, and GQ344828), four each from Merlot (GQ344815, GQ344816, GQ344817, and GQ344818) and Syrah (GQ344819, GQ344820, GQ344821, and GQ344822), and three from Lemberger (GQ344812, GQ344813, and GQ344814) showed 87 to 100% identity at the nucleotide level and 92 to 100% identity at the amino acid level. A pairwise comparison of HSP-70 sequences of GLRaV-3 isolates from Idaho with corresponding sequences of GLRaV-3 isolates from GenBank showed nucleotide sequence identities between 88% (AJ748519) and 100% (DQ780885). Phylogenetic analysis of HSP-70 sequences from Idaho and GenBank showed clustering of Idaho sequences into five groups, with 12 sequences clustering with a Washington isolate (DQ780885), six sequences in a second group clustering with an isolate from Tunisia (AJ748522), two sequences in a third group clustering with an isolate from Austria (AJ748513), and one sequence each in groups four and five clustering with isolates from Italy (AJ748520) and Washington (DQ780889), respectively. The clustering was not cultivar- or vineyard-specific, suggesting separate introductions of different GLRaV-3 isolates in planting materials. To our knowledge, this is the first report of GLRaV-3 in grapevines grown in Idaho. These and previous results (1,2), indicate the wide distribution of GLRaV-3 in several grapevine cultivars in the Pacific Northwest Region. References: (1) R. R. Martin et al. Plant Dis. 89:763, 2005. (2) R. A. Naidu et al. (Abstr.) Phytopathology 96(suppl.):S83, 2006. (3) M. J. Soule et al. Plant Dis. 90:1461, 2006.
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PMID:First Report of Grapevine leafroll-associated virus-3 in Six Wine Grape Cultivars in Idaho. 3075 10

Garlic (Allium sativum L.) is one of the most important Allium spp. plants that are widely cultivated throughout the world. A significant reduction in yield and quality due to virus infection is now a serious economic problem (1). In many cases, garlic plants are infected with a variety of viruses, but elimination of these viruses is difficult because this crop is propagated through bulbs. Potyviruses, carlaviruses, and allexiviruses have been detected in diseased garlic. Onion yellow dwarf virus (OYDV) and Leek Yellow Stripe Virus (LYSV), genus Potyvirus, family Potyviridae, are two important viral pathogens of garlic. Virus diseases of garlic are widespread in the world, causing serious damage to yields and quality of the crop. The East Mediterranean Region produces 14% of the garlic production of Turkey (110,000 t). A survey was done in garlic fields in Adana, Mersin, Kahramanmaras, Hatay, and Gaziantep provinces of Turkey where virus-like symptoms were noted in samples collected during the 2007-2008 growing season. Leaf and bulb samples were taken from 202 plants with leaf yellow stripe, mosaic, enations, and deformation or dwarfism symptoms. ELISA was performed with antibodies from Agdia (Elkhart, IN). Results indicated that 57 samples (28.2%) were infected with OYDV and 43 samples (21.2%) were infected with LYSV. In addition, 23 samples were determined to be infected by both viruses. All ELISA-positive samples and 10 ELISA-negative samples were analyzed by reverse transcription-PCR with primers 1OYDV-G (5' TTA CAT TCT AAT ACC AAG CA 3') and 2OYDV-G (5' GCA GGA GAT GGG GAG GAC GC 3') for the detection of OYDV and primers 1LYSV (5' TCA CTG CAT ATG CGC ACC AT 3') and 2LYSV (5' GCA CCA TAC AGT GAA TTG AG 3') for the detection of LYSV. These primers were previously reported to be specific for the coat protein genes of OYDV and LYSV, respectively (2). Products of the expected size (774 bp for OYDV and 1,020 bp for LYSV) were amplified only from the ELISA-positive samples of the respective viruses, confirming infections by OYDV and LYSV. To our knowledge, this is the first report of OYDV and LYSV in garlic in Turkey. References: (1) L. Bos et al. Neth. J. Plant Pathol. 84:185, 1978. (2) T. V. M. Fajardo et al. Fitopatol. Bras. 26:619, 2001.
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PMID:First Report of Onion yellow dwarf virus and Leek yellow stripe virus in Garlic in Turkey. 3076 28

Viruses are a major biotic constraint on sweet potato (Ipomoea batatas (L.) Lam) production worldwide. In 2005, 10 to 60% viral disease incidence was observed in sweet potato fields. Symptoms include ring and chlorotic spots, puckering, feathering, vein clearing, and leaf curl with chlorotic specks and pink spots. Cuttings from symptomatic plants were collected from Kerala (two clones), Orrisa (eight clones), and Adrapradesh (three clones) and maintained in an insect-proof glasshouse. Leaves from symptomatic plants were mechanically inoculated to I setosa, I. nil, Nicotiana tabacum, N. benthamiana, Datura stramonium, and Chenapodium quinoa (12 seedlings each). Vein clearing, netting, and leaf distortion were observed in I. setosa and N tabacum 7 days postinoculation, chlorotic spots observed in N. benthamiana, and violet spots and violet margins on leaves observed on I. Nil. No symptoms were observed on D. stramonium and C. quinoa. When scions from the symptomatic sweet potato plants were graft inoculated onto I. setosa, vein clearing, leaf curl, and puckering-like symptoms were observed within 5 days. Mosaic and leaf curling symptoms were also observed on mechanically inoculated N. tabacum. Total nucleic acids isolated from the 33 field-collected sweet potato samples, graft inoculated I. setosa plants, and mechanically inoculated N. tabacum and I. nil plants were used for PCR and reverse transcription (RT)-PCR with geminivirus group specific (2) and potyvirus group specific primers (1). The expected 530-bp and 1.3-kb fragment were generated from the geminivirus and potyvirus primer sets, respectively. Potyvirus alone was detected in 7 of the 33 field-collected plants; geminivirus alone was detected in 7 other plants, while 19 plants contained detectible levels of potyvirus and geminivirus. To further identify the viruses, nested primers specific for the coat protein gene of Sweet potato feathery mottle virus (SPFMV) (CP1S 5'AGT GGG AAG GCA CCA TAC ATA GC 3', CP1A5' GCA GAG GAT GTC CTA TTG CAC ACC 3') (CP2S 5'TCT AGT GAA CGT ACT GAA TTC AAA GA 3', CP2A 5'ATT GCA CAC CCC TGA TTC CTA AGA 3') and Sweet potato leaf curl virus (SPLCV) (CP1- 5'ATG ACA GGG CGA ATT CGC GTT TC 3', CP2- 5'TTA ATT TTT GTG CGA ATC ATA 3') were designed. I. setosa and N. tabacum were amplified with SPFMV and SPLCV primers and the amplicons of 960 and 764 bp, respectively, obtained were subsequently cloned into pGEM-T Easy vector and sequenced. Nucleotide BLAST analysis revealed that the 960-bp fragment (GenBank Accession No. EF015398.) was 98% identical to two Egyptian isolates of SPFMV (Nos. AJ 515379 and AJ 515378). The nucleotide sequence of the 764-bp products (Nos. EF 151926 and EF15483) from the samples collected from Kerala and Orisa was 95% identical to each other. The sequence identity of EF 15483 with Sweet potato leaf curl Georgia virus (SPLCGV) isolate AF326775. was 91% and identity with China isolate DQ 512731 was 90% The isolate EF 151926 also was 91% identical to the SPLCGV with a high query and alignment score whereas identity with the China isolate was 91% with a low query coverage and alignment score. Phylogenic analysis with MEGA software program also showed the highest sequence similarity with SPLCGV, hence it is concluded that the geminivirus isolate under study is SPLCGV. To our knowledge, this is the first report of identification of SPFMV and SPLCGV occurring on sweet potato in India. Further study is required to understand the consequences of occurrence of these two viruses in India. References: (1) D. Colinet et al. Plant Dis. 28:223 1998. (2) D. D. Deng et al. Ann. Appl. Biol 125:327, 1993.
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PMID:Occurrence of Sweet potato feathery mottle virus and Sweet potato leaf curl Georgia virus on Sweet Potato in India. 3076 15

Dahlia (Dahlia variabilis Hort.) is a significant ornamental plant in New Zealand. Symptoms such as mosaic, ring spots, mottling, and veinal chlorosis, suggestive of a viral infection, are often seen in various dahlia collections. To better understand the incidence of viruses in dahlia in New Zealand, several popularly grown cultivars were evaluated for viruses that are known to infect dahlia. Viruses that were tested included Cucumber mosaic virus (CMV), Dahlia mosaic virus (DMV), Impatiens necrotic spot virus (INSV), Tobacco streak virus (TSV), and Tomato spotted wilt virus (TSWV). At least one symptomatic plant was tested from each of the following cultivars: Akito Dawn, Cincinnati Dancer, Hamari Accord, Hamari Rose, LeBatts Prime, LeVonne Splinter, Riverlea Tropicana, Spartacus, Tartan, Tui Connie, and Wandas Antartica. Except for DMV, initial testing was done by ELISA with commercially available kits for the above viruses. In the case of dahlia mosaic, samples were tested for DMV that was described previously (4) and two additional and distinct caulimoviruses (DMV-D10 and DMV-Holland) that were found to be associated with dahlia (1,2). Primer pairs, ORF6st: ATG GAA GAA ATT AAG GCG T and ORF6end: TTG TCT TCA TCC ATA AAG CAG; DenF1: CAG CAA GAA ACA GGA ATT GA and DenR: TTA CAG TCG AAG CTG CTA AA; and Kapht-F: ATG AGT AAT GCT TCA GCA A and Kapht-R: TGA CCA TGG CTT CTA ACT GT were used for the specific detection of DMV-D10, DMV-Holland, and DMV, respectively (1). None of the samples tested were ELISA positive for CMV, INSV, or TSWV. To verify the TSV infection, TSV-specific primers (5'-GTC CAG ACC ATC CAT CCA AC-3' and 5'-TTG ATT CAC CAG GAA ATC TT-3'), designed based on sequences available in GenBank, were used in reverse transcription (RT)-PCR. For DMV, the diagnostic tests used were electron microscopy and PCR followed by amplicon cloning and sequencing. Electron microscopic observation of leaf-dip preparations showed near isometric virions, approximately 50 to 60 nm in all samples tested. PCR showed that all samples tested were positive for DMV-Holland and DMV-D10. While DMV-Holland is a typical caulimovirus, DMV-D10 was found to exist as an endogenous plant pararetroviral sequence in dahlia (3). One sample each from two cultivars, Spartacus and Tui Connie, were positive for TSV by ELISA, RT-PCR, followed by the sequence analysis of the cloned amplicon. The impact of TSV-infected dahlias as a potential source of inoculum remains to be seen. Our results suggested the prevalence of dahlia mosaic-associated caulimoviruses in several dahlia cultivars and the presence of TSV in New Zealand dahlias. Dahlia mosaic continues to be prevalent in several parts of the world (1), and with the current findings in New Zealand, testing for these viruses should be conducted to ensure virus-free status of the propagating material. References: (1) V. Pahalawatta et al. Plant Dis. 91:1194, 2007. (2) V. Pahalawatta et al. Arch. Virol.153:733, 2008. (3) V. Pahalawatta et al. Virology 376:253, 2008. (4) R. D. Richins and R. J. Shepherd. Virology 124:208, 1983.
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PMID:Dahlia mosaic virus and Tobacco streak virus in Dahlia (Dahlia variabilis) in New Zealand. 3076 5

Washington State is the largest producer of juice grapes (Vitis labruscana 'Concord' and Vitis labrusca 'Niagara') and ranks second in wine grape production in the United States. Grapevine leafroll disease (GLD) is the most wide spread and economically significant virus disease in wine grapes in the state. Previous studies (2) have shown that Grapevine leafroll associated virus-3 (GLRaV-3) is the predominant virus associated with GLD. However, little is known about the incidence and economic impact of GLD on juice and table grapes. Because typical GLD symptoms may not be obvious among these cultivars, the prevalence and economic impact of GLD in Concord and Niagara, the most widely planted cultivars in Washington State, has received little attention from the grape and nursery industries. During the 2005 growing season, 32 samples from three vineyards and one nursery of 'Concord' and three samples from one nursery of 'Niagara' were collected randomly. Petiole extracts were tested by single-tube reverse transcription-polymerase chain reaction (RT-PCR; 3) with primers LC 1 (5'-CGC TAG GGC TGT GGA AGT ATT-3') and LC 2 (5'-GTT GTC CCG GGT ACC AGA TAT-3'), specific for the heat shock protein 70 homologue (Hsp70h gene) of GLRaV-3 (GenBank Accession No. AF037268). One 'Niagara' nursery sample and eleven 'Concord' samples from the three vineyards tested positive for GLRaV-3, producing a single band of the expected size of 546 bp. The 'Niagara' and six of the 'Concord' RT-PCR products were cloned in pCR2.1 (Invitrogen Corp, Carlsbad, CA) and the sequences (GenBank Accession Nos. DQ780885, DQ780886, DQ780887, DQ780888, DQ780889, DQ780890, and DQ780891) compared with the respective sequence of a New York isolate of GLRaV-3 (GenBank Accession No. AF037268). The analysis revealed that GLRaV-3 isolates from 'Concord' and 'Niagara' share nucleotide identities of 94 to 98% and amino acid identities and similarities of 97 to 98% with the Hsp70h gene homologue of the New York isolate of GLRaV-3. Additional testing by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) using antibodies specific to GLRaV-3 (BIOREBA AG, Reinach, Switzerland) further confirmed these results in the 'Niagara' and two of the 'Concord' isolates. GLRaV-3 has previously been reported in labrusca cvs. Concord and Niagara in western New York (4) and Canada (1), but to our knowledge, this is the first report of GLRaV-3 in American grapevine species in the Pacific Northwest. Because wine and juice grapes are widely grown in proximity to each other in Washington State and grape mealybug (Pseudococcus maritimus), the putative vector of GLRaV-3, is present in the state vineyards, further studies will focus on the role of American grapevine species in the epidemiology of GLD. References: (1) D. J. MacKenzie et al. Plant Dis. 80:955, 1996. (2) R. R. Martin et al. Plant Dis. 89:763, 2005. (3) A. Rowhani et al. ICGV, Extended Abstracts, 13:148, 2000. (4) W. F. Wilcox et al. Plant Dis. 82:1062, 1998.
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PMID:First Report of Grapevine leafroll associated virus-3 in American Vitis spp. Grapevines in Washington State. 3078 Sep 26