The albumin-bilirubin grade uncovers the prognostic relationship between hepatic reserve and immune dysfunction in HIV-associated hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is a leading cause of liver-related mortality in people living with HIV, where co-infection with hepatotropic viruses accelerates the course of chronic liver disease. AIM: To evaluate whether the albumin-bilirubin (ALBI) grade, a more accurate marker of liver dysfunction in HCC, might identify patients with progressive liver dysfunction in the context of HIV/hepatitis co-infection. METHODS: Using uni- and multi-variable analyses, we studied the albumin-bilirubin grade as a predictor of overall survival (OS) in a large, multi-center cohort of patients with HIV-associated HCC recruited from 44 centres in 9 countries within the Liver Cancer in HIV study group. Patients who underwent liver transplantation were excluded. RESULTS: A total of 387 patients, predominantly HCV co-infected (78%) with balanced representation of all Barcelona Clinic Liver Cancer (BCLC) stages (A = 33%, B = 18%, C = 37%, D = 12%) were recruited. At HCC diagnosis, 84% had been on anti-retrovirals for a median duration of 8.8 years. The albumin-bilirubin grade identified significant differences in median survival of 97 months for grade 1 (95% CI 13-180 months), 17 months for grade 2 (95% CI 11-22 months) and 6 months for grade 3 (95% CI 4-9 months, P < .001). A more advanced albumin-bilirubin grade correlated with lower CD4 counts (464/373/288 cells/mm3 for grades 1/2/3) and higher HIV viraemia (3.337/8.701/61.845 copies/mL for grades 1/2/3, P < .001). CONCLUSIONS: In this large, multi-center retrospective study, the albumin-bilirubin grade highlights the interplay between liver reserve and immune dysfunction as prognostic determinants in HIV-associated HCC.

A TOMM40 poly-T variant modulates gene expression and is associated with vocabulary ability and decline in nonpathologic aging.

The Translocase of Outer Mitochondrial Membrane 40 Homolog and Apolipoprotein E (TOMM40-APOE) locus has been associated with a number of age-related phenotypes in humans including nonpathologic cognitive aging, late-onset Alzheimer's disease, and longevity. Here, we investigate the influence of the TOMM40 intron 6 poly-T variant (rs10524523) on TOMM40 gene expression and cognitive abilities and decline in a cohort of 1613 community-dwelling elderly volunteers who had been followed for changes in cognitive functioning over a period of 14 years (range = 12-18 years). We showed that the shorter length poly-T variants were found to act as a repressor of luciferase gene expression in reporter gene constructs. Expression was reduced to approximately half of that observed for the very long variant. We further observed that the shorter poly-T variant was significantly associated with reduced vocabulary ability and a slower rate of vocabulary decline with age compared to the very long poly-T variants. No significant associations were observed for memory, fluid intelligence or processing speed, although the direction of effect, where the short variant was correlated with reduced ability and slower rate of decline was observed for all tests. Our results indicate that the poly-T variant has the ability to interact with transcription machinery and differentially modulate reporter gene expression and influence vocabulary ability and decline with age.

Decreased defense gene expression in tolerance versus resistance to Verticillium dahliae in potato.

Verticillium dahliae Kleb., a soil-borne fungus that colonizes vascular tissues, induces wilting, chlorosis and early senescence in potato. Difference in senescence timing found in two diploid potato clones, 07506-01 and 12120-03, was studied and genetic variation in response to V. dahliae infection was identified as a causal factor. The clone, 07506-01, was infected with V. dahliae but did not develop symptoms, indicating tolerance to the pathogen. The other diploid clone, 12120-03 had low levels of pathogen with infection and moderate symptoms indicating partial resistance. 07506-01 was found to carry two susceptible alleles of the Ve2 gene and 12120-03 carried one Ve2 resistant and one susceptible allele. Infected leaves of the two clones were compared using gene expression profiling with the Potato Oligonucleotide Chip Initiative (POCI) microrarray. The results provide further evidence for differences in response of the two clones to infection with V. dahliae. Chlorophyll biosynthesis was higher in the tolerant 07506-01 compared to partially resistant 12120-03. On the other hand, expression of fungal defense genes, Ve resistance genes and defense phytohormone biosynthetic enzyme genes was decreased in 07506-01 compared to 12120-03 suggesting defense responses were suppressed in tolerance compared to resistance. Transcription factor gene expression differences pointed to the WRKY family as potential regulators of V. dahliae responses in potato.

Molecular phylogeny of Boliniales (Sordariomycetes) with an assessment of the systematics of Apiorhynchostoma, Endoxyla and Pseudovalsaria.

The systematics of the ascomycete genera Apiorhynchostoma, Endoxyla and Pseudovalsaria are reevaluated based on the comparison of cultural characteristics, teleomorph morphology and DNA sequence data. Analyses of sequences of the internal transcribed spacer (ITS) of the ribosomal DNA operon and the large subunit (LSU) of the nuclear ribosomal DNA gene resolve Boliniales as a robustly supported lineage comprising Apiorhynchostoma, Camarops, Camaropella, Cornipulvina, Endoxyla and Pseudovalsaria. Within Boliniales, species of Endoxyla form a strongly supported lineage. Apiorhynchostoma curreyi and Pseudovalsaria ferruginea group with Cornipulvina ellipsoides. Species of Camarops are paraphyletic and comprise two clades, one of which includes Camaropella. Boliniaceae is emended, Endoxyla mallochii is described as new and Apiorhynchostoma trabicola is considered a synonym of Apiorhynchostoma altipetum. We also propose the combinations Endoxyla occulta, Endoxylina luteobasis and Jobellisia peckii. Keys to genera included in the Boliniaceae and to species of Apiocamarops, Apiorhynchostoma and Endoxyla are provided.

Genetic Composition of Phytophthora infestans in Canada Reveals Migration and Increased Diversity.

A dramatic increase in the incidence of late blight and changes within populations of Phytophthora infestans have been observed in various regions of Canada. In this study, the occurrence of several new genotypes of the pathogen was documented with associated phenotypes that dominated pathogen populations. Genotype US-23, previously detected only among isolates from the United States, dominated in the western Canadian provinces of British Columbia, Alberta (AB), Saskatchewan, and Manitoba (MB). Although isolates of US-23 infect both potato and tomato, these isolates were the only genotype recovered from commercial garden centers in Canada. Isolates of genotype US-8, previously dominant throughout Canada, represented the only genotype detected from the eastern Canadian provinces of New Brunswick and Prince Edward Island. Isolates of other genotypes detected in Canada included US-11 in AB, US-24 in MB, and US-22 in Ontario (ON). An additional genotype was detected in ON which appears to be a derivative of US-22 that may have arisen through sexual reproduction. However, evidence of clonal reproduction dominated among the isolates collected, and opportunities for sexual reproduction were probably limited because of a surprising geographic separation of the A1 and A2 mating types in Canada. Sensitivity of the US-22, US-23, and US-24 isolates to the fungicide metalaxyl, movement of potato seed and transplants, and weather conditions may have contributed to reduced opportunities for contact between the mating types in fields in Canada. All P. infestans isolates were readily distinguished from other related oomycetes with RG57 restriction fragment length polymorphism analysis. Long-distance movement in seed tubers and garden center transplants may have contributed to the rapid spread of the P. infestans genotypes across Canada. Tracking pathogen movement and population composition should improve the ability to predict the genotypes expected each year in different regions of Canada.

The protein tyrosine phosphatase N22 gene is associated with juvenile and adult idiopathic inflammatory myopathy independent of the HLA 8.1 haplotype in British Caucasian patients.

OBJECTIVE: To examine single-nucleotide polymorphisms (SNPs) of the protein tyrosine phosphatase N22 gene (PTPN22) and to study the relationship between PTPN22 and the HLA region in patients with idiopathic inflammatory myopathies (IIMs). METHODS: PTPN22 SNPs were assessed in a large, cross-sectional, case-control study from the UK involving patients with adult or juvenile IIM, comprising patients with polymyositis (PM) (n=114), dermatomyositis (DM) (n=102), myositis associated with another connective tissue disease (myositis-CTD overlap syndrome) (n=64), or juvenile DM (n=101), in comparison with 748 control subjects. Seventeen PTPN22 SNPs were genotyped using the Sequenom MassArray iPLEX platform. Serotyping for myositis-specific/myositis-associated autoantibodies (MSAs/MAAs) was performed by radioimmunoprecipitation. RESULTS: A significant association was noted between the R620W variant (rs2476601) and IIM (corrected P [Pcorr]=0.0009 versus controls), and specifically with the clinical subgroup of PM (Pcorr=0.003 versus controls). A weaker association was noted with juvenile DM (Pcorr=0.009 versus controls). No significant associations were noted after stratification by serologic subgroups. The association with the R620W variant was independent of alleles forming the HLA 8.1 haplotype. No other PTPN22 SNPs were associated with IIM. The PTPN22 haplotype containing the R620W T allele was the only haplotype significantly associated with IIM. CONCLUSION: The R620W variant is a significant risk factor for IIM, independent of the HLA 8.1 haplotype. Unlike that in the HLA region, risk is not increased in individuals possessing MSAs/MAAs. These results are further evidence that the PTPN22 gene confers autoimmune susceptibility.

EBV Immortalization of human B lymphocytes separated from small volumes of cryo-preserved whole blood.

EBV immortalized B lymphocyte cell lines have been extensively used as a source of biological material for functional and molecular studies and represent a potentially limitless source of genomic DNA. Current technologies for EBV transformation are costly and use relatively large volumes of peripheral blood. Alternative methods were examined to determine whether smaller volumes of cryo-preserved whole blood could be subsequently transformed and which could provide a more cost-effective strategy for large population-based studies such as UK Biobank. A successful method was established where viable B cells were positively selected from 0.5 ml cryo-preserved whole blood samples. These were EBV transformed in microtitre plates and subsequently expanded in culture. A pilot study within UK Biobank was performed, which confirmed its potential usefulness for this study.

First Report of Fludioxonil-Resistant Isolates of Fusarium spp. Causing Potato Seed-Piece Decay.

Potato (Solanum tuberosum L.) diseases incited by Fusarium spp. include postharvest dry rot and seed-piece decay. Fusarium seed-piece decay is commonly controlled by preplant applications of chemical seed treatments. However, isolates of Fusarium spp. resistant to benzimidazole fungicides have been reported (2,4). In the spring of 2007, samples of cut seed tubers (cvs. Shepody and Russet Burbank) showing extensive symptoms of decay were received from three seedlots in Prince Edward Island (PE) and one seedlot in Saskatchewan (SK), Canada. All seed tubers had been treated with fludioxonil (Maxim Potato Seed Protectant [PSP], 0.5% fludioxonil) following cutting and then stored for 10 to 14 days prior to planting. Using standard isolation protocols (4), the 19 potato tuber pieces examined from PE and 2 from SK yielded 21 Fusarium isolates for further study. Five isolates (including both isolates from SK) were identified as Fusarium sambucinum Fuckel and the remaining 16 isolates were identified as F. coeruleum (Libert) Sacc. (3). To confirm identifications, isolates were compared with two known standards of each of F. sambucinum and F. coeruleum identified by K. Seifert (Agriculture and Agri-Food Canada, Ottawa, ON) by DNA sequencing of the partial ß-tubulin gene or the translation elongation factor 1-α ( http://fusarium.cbio.psu.edu ; [1]). These standard isolates were also used as fludioxonil-sensitive controls in amended agar assays for chemical sensitivity. Agar plugs (5 mm in diameter) taken from the margins of 7-day-old cultures of the Fusarium isolates were transferred to petri dishes containing ½-strength potato dextrose agar amended with 0, 0.1, 1.0, 10.0, or 100.0 mg/liter of fludioxonil. Fludioxonil (Maxim PSP, 0.5% a.i.) was prepared as a stock solution in sterile distilled water and added to the molten agar after autoclaving. Culture incubation and mycelial growth measurements were performed as described previously (4). Measurements from four replicate petri dishes per concentration of fludioxonil were taken. Calculated EC50 values (fludioxonil concentration inhibiting pathogen growth by 50%) were obtained. The trial was repeated three times. The two standard isolates of F. sambucinum were sensitive to fludioxonil, with mean EC50 values of 0.002 (±0.002 standard error [SE]) and 0.005 (±0.002 SE) mg/liter. The two standard isolates of F. coeruleum were also sensitive to fludioxonil, with mean EC50 values of 0.17 (±0.005 SE) and 0.19 (± 0.005 SE) mg/liter. All other tested isolates of F. sambucinum and F. coeruleum were resistant to fludioxonil and showed no growth inhibition even at 100 mg of fludioxonil per liter. To our knowledge, this is the first report of resistance to fludioxonil in isolates of Fusarium spp. causing potato seed-piece decay. Since the isolates of F. sambucinum were also resistant to thiophanate-methyl and thiabendazole (data not shown), multiclass (benzimidazole and pyrrole) resistance was also documented. References: (1) D. M. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004. (2) L. M. Kawchuk et al. Am. Potato J. 71:185, 1994. (3) P. E. Nelson et al. Fusarium Species: An Illustrated Manual for Identification. Pennsylvania State University Press, 1983. (4) R. D. Peters et al. Plant Dis. 85:1030, 2001.

In-Furrow Applications of Metalaxyl and Phosphite for Control of Pink Rot (Phytophthora erythroseptica) of Potato in New Brunswick, Canada.

The efficacy of metalaxyl-m (Ridomil Gold 480EC) and phosphite (Phostrol) applied at planting in-furrow against pink rot (Phytophthora erythroseptica) of potato (Solanum tuberosum) 'Shepody' and 'Russet Burbank' was evaluated in field trials conducted in 2005 and 2006 in Florenceville, New Brunswick, Canada. Inoculum made from a metalaxyl-m-sensitive isolate of P. erythroseptica from New Brunswick was applied either in-furrow as a vermiculite slurry at planting or as a zoospore drench in soils adjacent to potato plants in late August. After harvest, the number and weight of tubers showing pink rot symptoms were assessed and expressed as percentages of the total tuber number and total weight of tubers. Metalaxyl-m applied in-furrow was significantly more effective against pink rot than phosphite. The mean percentage of diseased tubers as a percentage of total tuber weight was 1.5% (2005) and 1.2% (2006) for metalaxyl-m-treated plots and 9.6% (2005) and 2.8% (2006) for phosphite-treated plots, a percentage similar to that obtained in inoculated control plots with no fungicide treatment. The mean percentage of diseased tubers expressed as a percentage of the total number of tubers was 1.7% (2005) and 1.3% (2006) for metalaxyl-m-treated plots and 10.1% (2005) and 3.1% (2006) for phosphite-treated plots. Disease incidence was significantly higher using the late-season inoculation technique (respective means in 2005 and 2006 were 9.9 and 3.8% diseased tubers, by weight, and 10.6 and 3.9%, by number) than with the in-furrow inoculation method (respective means in 2005 and 2006 were 3.3 and 0.7% by weight, and 3.7 and 1.3%, by number). The potato cv. Shepody was significantly more susceptible to pink rot (9.9 and 3.3% diseased tubers, by weight, in 2005 and 2006, respectively, and 10.6 and 3.9%, by number) than Russet Burbank (respective means in 2005 and 2006 were 3.4,% and 1.2%, by weight, and 3.7,% and 1.2%, by number). Our findings indicate that metalaxyl applied in-furrow at planting is a viable option for control of pink rot caused by metalaxyl-sensitive strains of P. erythroseptica, whereas phosphite was ineffective.

Polymorphisms in the tumour necrosis factor gene are not associated with severity of inflammatory polyarthritis.

BACKGROUND: Tumour necrosis factor alpha (TNFalpha) is a powerful inflammatory mediator in rheumatoid and other types of inflammatory arthritis. Polymorphisms within the TNFalpha gene have previously been investigated to determine their role in the aetiopathogenesis of rheumatoid arthritis (RA), but it is unclear whether reported associations are with susceptibility to, or severity of, disease. OBJECTIVE: To examine the association between both individual TNFalpha single nucleotide polymorphisms (SNPs) and haplotypes with the development and severity of erosions by 5 years in patients with inflammatory polyarthritis (IP). METHODS: 438 patients from the Norfolk Arthritis Register observational inception cohort of patients with IP were x rayed 5 years after disease onset. They were genotyped for nine SNPs mapping to the TNFalpha gene, using a SNaPshot primer extension assay. Haplotypes were constructed in patients with IP, who were compared for the presence and extent of erosions at 5 years. RESULTS: No association between individual TNFalpha SNPs or haplotypes in the patients who developed erosions at 5 years compared with those who remained non-erosive was found. Restricting analysis to patients who satisfied ACR criteria for RA by 5 years did not affect the conclusions. CONCLUSION: The TNFalpha gene does not seem to be associated with severity as assessed by erosive outcome at 5 years in patients with IP.

Molecular evidence that Verticillium ablo-atrum Grp 2 isolates are distinct from V. albo-atrum Grp 1 and V. tricorpus.

summary Verticillium species-specific primers were used in a polymerase chain reaction to differentiate between Verticillium albo-atrum groups and V. tricorpus. Amplification with species-specific primers identified 21 isolates from the 64 V. albo-atrum isolates tested as Grp2. Genome analysis using RAPDs and restriction fragment length polymorphism (RFLP) of the intergenic (IGS) region of the rDNA showed that V. albo-atrum Grp2 isolate were genetically distinct from either V. albo-atrum Grp1 or V. tricorpus, demonstrating a significant differentiation between these species. The sizes of the amplified IGS fragment were different, with Grp1 isolates having a smaller fragment ( approximately 2.1 kb) than either Grp2 ( approximately 2.3 kb) or V. tricorpus ( approximately 2.7 kb). Based on RAPD analysis, the average similarity coefficients between Grp1 and Grp2 were 35% and 34% between Grp2 and V. tricorpus. Multiple correspondence analysis separated the isolates into three major groups corresponding to Grp1, Grp2 and V. tricorpus. Surprisingly, isolates collected from Pisum sativa were distinct from other Grp1 V. albo-atrum isolates. The observed low levels of genetic similarity, the differences in sizes of IGS fragments, IGS-RFLP profiles and the RAPD patterns point to the possibility of Grp2 isolates comprising a different species of Verticillium than those occupied by either V. albo-atrum Grp1 isolates or V. tricorpus.

Impulsiveness, serotonin genes and repetition of deliberate self-harm (DSH).

BACKGROUND: Few studies have investigated independent associations of psychological, biological and social variables with repeated deliberate self-harm (DSH). Serotonin function has been linked to impulsive and suicidal behaviour and genetic polymorphisms have been identified within the serotonin system that could account for this link. This study tested hypotheses linking impulsiveness, genetic polymorphisms of tryptophan hydroxylase (TPH) and the 5-HT2c receptor and repeated DSH. METHODS: Individuals presenting after DSH were interviewed, completed personality questionnaires and gave venous blood samples. Genotypes were determined for TPH intron7 and 5-HT2c (cys-ser) polymorphisms. Follow-up to identify repetition of DSH was for 1 year. RESULTS: Males with the 5-HT2c serine variant were more impulsive than those with the cysteine variant (039 standardized units, P = 0.041, 95% CI 0.017 to 0.076). There was no association between impulsiveness and the TPH intron7 polymorphism overall but a weak association with the L allele in men (0.41 standardized units, P = 0.05, 95 % CI 0.001 to 0.82). Impulsiveness, although high in the group as a whole, did not distinguish those who repeated DSH. CONCLUSIONS: The personality trait of impulsiveness may in part be related to genotypes of the 5-HT2c receptor and TPH gene in men. Impulsiveness does not differ between those who do and do not repeat DSH.

Variation in Aggressiveness of Canadian Isolates of Phytophthora infestans as Indicated by Their Relative Abilities to Cause Potato Tuber Rot.

Studies were conducted in 1995, 1996, and 1997 to determine the ability of Canadian isolates of Phytophthora infestans to cause tuber disease in a variety of potato cultivars. Most isolates of recently introduced multilocus genotypes (US-7, US-8, g11, g26, g29, and g40) were more aggressive on tuber tissue than isolates of the traditional US-1 genotype, based on surface necrosis (SN), lesion depth (LD), and compound aggressiveness index (CAI = SN × LD) components. Other multilocus genotypes (g30, g41, g42, and UN-3) were similar in aggressiveness to US-1. The g11 (US-11) genotype consistently displayed aggressiveness that was intermediate to that of the US-8 and US-1 genotypes. Variation also occurred within a multilocus genotype, and an isolate of the US-8 genotype from New Brunswick was consistently less aggressive than other US-8 isolates. Cvs. Dorita and Island Sunshine were consistently the most resistant to infection, regardless of isolate, relative to the more susceptible responses of Green Mountain, Russet Bur-bank, Kennebec, Superior, Shepody, Red Pontiac, Sebago, and Bintje. Even so, isolates of the newly introduced US-8 genotype were able to cause significantly more disease on these resistant cultivars than isolates of the US-1 genotype. The predominant genotypes currently found in potato production areas in Canada (US-8 and g11) have higher fitness on tuber tissue than displaced, preexisting strains (US-1).

Comparison of three solid phase supports for promoting three-dimensional growth and function of human liver cell lines.

An extracorporeal liver support system will require that liver cells maintain their normal differentiated function. This is more likely to be achieved utilizing a three-dimensional culture configuration rather than a simple monolayer culture. We present data on a human liver cell line attached and maintained on different three-dimensional supports, porous glass (Siran), silicon (Immobasil), and calcium-alginate beads. Albumin, fibrinogen, prothrombin, alpha1-acid glycoprotein and alpha1-antitrypsin secretions were measured. Proliferation was slower on each of the three-dimensional supports than on the monolayer culture. The protein secretion of all 5 proteins was highest in cells encapsulated in alginate; silicon beads supported greater protein secretion than glass. Cells on silicon or within alginate were rounded; those on glass grew in 2 configurations as flattened monolayers and as rounded colonies. Cells in alginate secreted as much protein as the whole liver (e.g., albumin, 14.88 g/10(12) cells/day compared to the whole liver, approximately 12 g/day). Three-dimensional culture of a human liver cell line leads to both proliferation and a high synthetic capacity, an important feature of cells suitable for an extracorporeal liver support system.

Outcome study using an administrative database: terminal salpingostomy, physician case load and live birth rates.

Information on the outcome following salpingostomy performed for infertility in everyday practice is needed to counsel patients, determine the best approach to this condition and clarify whether the outcome is superior when surgery is performed by a physician who maintains a high volume of ongoing experience. A total of 547 consecutive subjects were identified over a 5 year period using the Alberta Health Care Claims Database. Their claims history was analysed over a follow-up of 2-7 years, to identify pregnancy-related events, loss to follow-up and events which would result in sterility. Pregnancies were cross-checked with the only in-vitro fertilization programme serving the region. The overall cumulative live birth and tubal pregnancy rates were 11.7 and 7.2% respectively. Live birth rates were significantly higher when practitioners had performed >10 procedures within the study period. Live birth rates would appear to be substantially lower in everyday practice than following surgery performed by acknowledged experts. A high volume of ongoing experience appears to be associated with superior live birth rates. It is unclear whether this association relates to case selection, surgical expertise or both variables.