Publications by authors named "R A Fullerton"

28 Publications

Acadl-SNP based genotyping assay for long-chain acyl-CoA dehydrogenase deficient mice.

Mol Genet Metab 2012 May 15;106(1):62-7. Epub 2012 Feb 15.

Metabolic Signaling and Disease Program, Diabetes and Obesity Research Center, Sanford-Burnham Medical Research Institute at Lake Nona, Orlando, FL 32827, USA.

The long-chain acyl-CoA dehydrogenase (LCAD) (Acadl=gene; LCAD=protein) deficient mouse model has been important in evaluating the role of mitochondrial fatty acid oxidation of long-chain fatty acids in metabolic disorders. The insertion vector-based gene targeting strategy used to generate this model has made it difficult to distinguish homozygous and heterozygous genotypes containing targeted Acadl alleles in LCAD-deficient mice. Herein, we describe the design and validation of Acadl SNP genotyping methods capable of distinguishing between heterozygous and homozygous LCAD-deficient mice. The Acadl SNP genotyping assays are effective at allelic discrimination of both C57BL/6 and 129 mouse strain-based Acadl alleles under conditions including, both low purity and quantity genomic DNA templates. This makes the method practical and provides the necessary tools for genotyping the LCAD-deficient mouse model.
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http://dx.doi.org/10.1016/j.ymgme.2012.02.009DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3335976PMC
May 2012

Probing eudesmane cation-π interactions in catalysis by aristolochene synthase with non-canonical amino acids.

J Am Chem Soc 2011 Sep 11;133(35):13906-9. Epub 2011 Aug 11.

School of Chemistry, Cardiff University, Park Place, Cardiff, United Kingdom.

Stabilization of the reaction intermediate eudesmane cation (3) through interaction with Trp 334 during catalysis by aristolochene synthase from Penicillium roqueforti was investigated by site-directed incorporation of proteinogenic and non-canonical aromatic amino acids. The amount of germacrene A (2) generated by the mutant enzymes served as a measure of the stabilization of 3. 2 is a neutral intermediate, from which 3 is formed during PR-AS catalysis by protonation of the C6,C7 double bond. The replacement of Trp 334 with para-substituted phenylalanines of increasing electron-withdrawing properties led to a progressive accumulation of 2 that showed a good correlation with the interaction energies of simple cations such as Na(+) with substituted benzenes. These results provide compelling evidence for the stabilizing role played by Trp 334 in aristolochene synthase catalysis for the energetically demanding transformation of 2 to 3.
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http://dx.doi.org/10.1021/ja205927uDOI Listing
September 2011

First Record of Bacterial Crown Rot of Papaya (Carica papaya) Caused by an Erwinia papayae-Like Bacterium in the Kingdom of Tonga.

Plant Dis 2011 Jan;95(1):70

Landcare Research, Private Bag 92170, Auckland Mail Centre, Auckland 1142, New Zealand.

Symptoms resembling papaya bacterial crown rot (1,3) attributed to Erwinia papayae were first observed on 'Waimanalo' and 'Solo Sunrise' papaya on the island of Tongatapu, Kingdom of Tonga in July 2009. Spreading, dark green, water-soaked lesions formed on juvenile stem tissue and developed into a foul-smelling, wet rot that destroyed large sections of the stem. Coalescing, brown, angular, marginal, and intercostal lesions killed large areas of the lamina. Elongated lesions on petioles resulted in breakage and leaf death. Symptoms on stems typically moved toward the crown with the growing point being killed or the whole crown breaking off at a canker below. Isolations at 28°C on King's medium B (KB) yielded slow-growing, raised, white, mucoid colonies that produced a conspicuous, nondiffusable blue pigment in the medium. Two-day-old suspensions (1 × 10 CFU/ml) of two cultures were injected into juvenile stem tissue, petioles, and laminae of four healthy papaya seedlings of 'Solo Sunrise' with a sterile 1-ml insulin syringe. Sterile water was used as a negative control. Typical water-soaked lesions appeared at all bacterial inoculation sites on all plants within 5 days but not on controls. Pigment-producing colonies similar to those used for inoculation were reisolated from four different stem lesions. Bacteria isolated from diseased tissues were gram negative, facultative anaerobes, oxidase negative, nonfluorescent on KB, induced a hypersensitive reaction on tobacco leaves, but could not cause soft rot on potato slices. Those characteristics, combined with blue pigment production, are consistent with the bacterium E. papayae. A partial sequence of the 16S rDNA gene of ~804 bp was amplified from four Tongan isolates (ICMP18248-18251) using primers 27f and 1492r (4). Sequences of these strains were 100% identical to each other (GenBank Nos. HQ286366-HQ286369), 99 and 98% identical to the 16SrDNA sequences of E. mallotivora strains LMG2708 (Z96084.1) and DSM4565 (AJ233414.1) respectively, and 97% identical to the 16SrDNA sequence of E. papayae strain NCPPB 4294 (AY131237.1). E. mallotivora and E. papayae cause different diseases, a leaf spot on Mallotus japonicus (2) and bacterial canker on papaya, respectively. They are closely related and in the laboratory are distinguished by only very few biochemical characteristics (1). E. papayae produces a blue pigment on KB and can utilize arabinose but not mannitol. E. mallotivora does not produce a blue pigment and can utilize mannitol but not arabinose. The four Tongan strains produced a blue pigment and could utilize mannitol and arabinose. Symptoms caused by the strains isolated from Tonga are typical of those caused by E. papayae and the strains identified share most of the characteristics of E. papayae. Because the Tongan strains were able to utilize mannitol as well as arabinose and their 16S rDNA was only 97% similar to E. papayae, these strains are referred to as an E. papayae-like bacterium. The taxonomic position of these isolates will be resolved with techniques such as Multilocus Sequence Typing analysis. To our knowledge, this is the first report of this highly destructive papaya disease in the Kingdom of Tonga and of a pathogen closely related to E. papaya in the country. References: (1) L. Gardan et al. Int. J. Syst. Bacteriol. 54:107, 2004. (2) M. Goto. Int. J. Syst. Bacteriol. 26:467, 1976. (3) N. H. Maktar et al. New Dis. Rep. 17:4, 2008. (4) F. Martin-Laurent et al. Appl. Environ. Microbiol. 67:2354, 2001.
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http://dx.doi.org/10.1094/PDIS-06-10-0455DOI Listing
January 2011

Breast cancer patients' clinical outcome measures are associated with Src kinase family member expression.

Br J Cancer 2010 Sep 17;103(6):899-909. Epub 2010 Aug 17.

Western Infirmary Glasgow, Section of Surgery, Division of Cancer Sciences and Molecular Pathology, Faculty of Medicine, Level 2, McGregor Building, Dumbarton Road, Glasgow G11 6NT, UK.

Background: This study determined mRNA expression levels for Src kinase family (SFK) members in breast tissue specimens and assessed protein expression levels of prominent SFK members in invasive breast cancer to establish associations with clinical outcome. Ki67 was investigated to determine association between SFK members and proliferation.

Methods: The mRNA expression levels were assessed for eight SFK members by quantitative real-time PCR. Immunohistochemistry was performed for c-Src, Lyn, Lck and Ki67.

Results: mRNA expression was quantified in all tissue samples. SRC and LYN were the most highly expressed in malignant tissue. LCK was more highly expressed in oestrogen receptor (ER)-negative, compared with ER-positive tumours. High cytoplasmic Src kinase protein expression was significantly associated with decreased disease-specific survival. Lyn was not associated with survival at any cellular location. High membrane Lck expression was significantly associated with improved survival. Ki67 expression correlated with tumour grade and nuclear c-Src, but was not associated with survival.

Conclusions: All eight SFK members were expressed in different breast tissues. Src kinase was highest expressed in breast cancer and had a negative impact on disease-specific survival. Membrane expression of Lck was associated with improved clinical outcome. High expression of Src kinase correlated with high proliferation.
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http://dx.doi.org/10.1038/sj.bjc.6605829DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2966624PMC
September 2010

First Report of Verticillium Wilt of Gold Kiwifruit, Actinidia chinensis Cv. Hort 16A, Caused by Verticillium albo-atrum in Chile.

Plant Dis 2009 May;93(5):553

Departamento de Sanidad Vegetal, Facultad de Ciencias Agronómicas, Universidad de Chile, Santiago, Chile.

Gold kiwifruit, Actinidia chinensis Planch cv. Hort 16A, was first planted in Chile in 2003 and vines started dying within 2 years. By the end of the 2007-2008 growing season, as much as 80% of the plants in several orchards had died. The disease was characterized by a conspicuous reddish brown discoloration of the xylem and the sudden wilting and dieback of plants any time during the growing season. In the spring, entire plants or parts of plants failed to break buds. In others, the buds broke, but juvenile leaf clusters then wilted and died. On severely affected plants, scion watershoots wilted and died. The disease was often accompanied by shallow cracking of the bark and slight sponginess of the underlying cortex. The disease was apparently most severe in sites that had been planted to Gold kiwifruit immediately after removal of apple, pear, citrus, or grape. Orchards planted following long-term maize, wheat, or grass culture were almost disease free. A fungus was consistently isolated from symptomatic vascular tissue disinfected in 1% sodium hypochlorite and plated on potato dextrose agar. Conidiogenous cells were arranged in verticels; conidia were hyaline, elliptical, single celled, and measured 3.5 to 8.5 × 1.8 to 4.3 μm (average 5.5 × 2.5 μm). Dark, resting mycelium developed after 1 to 2 weeks of incubation. On the basis of these morphological characteristics, the fungus was identified as Verticillium albo-atrum Reinke & Berthier. Identification was confirmed by sequencing part of the internal transcribed spacer (ITS) region with primers ITS1 and ITS4. The sequence of a representative isolate showed high homology (98% identity over a length of 494 bp) with a DNA fragment (NCBI Accession No. 108476) of V. albo-atrum from alfalfa. To complete pathogenicity tests, 20 healthy, 1-year-old Hort 16A kiwi vines grafted on Hayward kiwifruit (A. deliciosa Chevalier) seedlings were inoculated by injection of 20 μl of 10 conidia/ml into stems of the scion. Twenty control plants were injected with an equal volume of sterile distilled water. Plants were held in a controlled environment facility at 24°C with 16 h of light per day. Eight weeks after inoculation, typical wilting and dieback symptoms developed on 90% of the plants. Control plants injected with water remained healthy. Verticillium wilt has never been reported on kiwifruit (A. deliciosa) in Chile. V. albo-atrum has a rather narrow host range and is mainly reported as a pathogen on alfalfa, hop, soybean, tomato, and potato (1). To our knowledge, this is the first report of V. albo-atrum causing wilt and dieback on Gold kiwifruit (A. chinensis) cv. Hort 16A. The fungal isolates have been deposited in the Plant Pathology Laboratory of the Sanidad Vegetal Department of Agricultural Sciences Faculty of University of Chile under the name Actinidia chinensis/V. albo-atrum No. 1 to 8. Reference: (1) E. K. Ligoxigakis et al. Phytoparasitica 30:511, 2002.
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http://dx.doi.org/10.1094/PDIS-93-5-0553BDOI Listing
May 2009