Risk Factors for De Novo Malignancy Following Lung Transplantation.

Risk factors for non-skin cancer de novo malignancy (DNM) after lung transplantation have yet to be identified. We queried the United Network for Organ Sharing database for all adult lung transplant patients between 1989 and 2012. Standardized incidence ratios (SIRs) were computed by comparing the data to Surveillance, Epidemiology, and End Results Program data after excluding skin squamous/basal cell carcinomas. We identified 18 093 adult lung transplant patients; median follow-up time was 1086 days (interquartile range 436-2070). DNMs occurred in 1306 patients, with incidences of 1.4%, 4.6%, and 7.9% at 1, 3, and 5 years, respectively. The overall cancer incidence was elevated compared with that of the general US population (SIR 3.26, 95% confidence interval [CI]: 2.95-3.60). The most common cancer types were lung cancer (26.2% of all malignancies, SIR 6.49, 95% CI: 5.04-8.45) and lymphoproliferative disease (20.0%, SIR 14.14, 95% CI: 9.45-22.04). Predictors of DNM following lung transplantation were age (hazard ratio [HR] 1.03, 95% CI: 1.02-1.05, p < 0.001), male gender (HR 1.20, 95% CI: 1.02-1.42, p = 0.03), disease etiology (not cystic fibrosis, idiopathic pulmonary fibrosis or interstitial lung disease, HR 0.59, 95% CI 0.37-0.97, p = 0.04) and single-lung transplantation (HR 1.64, 95% CI: 1.34-2.01, p < 0.001). Significant interactions between donor or recipient smoking and single-lung transplantation were noted. On multivariable survival analysis, DNMs were associated with an increased risk of mortality (HR 1.44, 95% CI: 1.10-1.88, p = 0.009).

Simulated Regionalization of Heart and Lung Transplantation in the United States.

We simulated the impact of regionalization of isolated heart and lung transplantation within United Network for Organ Sharing (UNOS) regions. Overall, 12 594 orthotopic heart transplantation (OHT) patients across 135 centers and 12 300 orthotopic lung transplantation (OLT) patients across 67 centers were included in the study. An algorithm was constructed that "closed" the lowest volume center in a region and referred its patients to the highest volume center. In the unadjusted analysis, referred patients were assigned the highest volume center's 1-year mortality rate, and the difference in deaths per region before and after closure was computed. An adjusted analysis was performed using multivariable logistic regression using recipient and donor variables. The primary outcome was the potential number of lives saved at 1 year after transplant. In adjusted OHT analysis, 10 lives were saved (95% confidence interval [CI] 9-11) after one center closure and 240 lives were saved (95% CI 209-272) after up to five center closures per region, with the latter resulting in 1624 total patient referrals (13.2% of OHT patients). For OLT, lives saved ranged from 29 (95% CI 26-32) after one center closure per region to 240 (95% CI 224-256) after up to five regional closures, but the latter resulted in 2999 referrals (24.4% of OLT patients). Increased referral distances would severely limit access to care for rural and resource-limited populations.

Prolonged Barium-Impaction Ileus in Two Lung Transplant Recipients With Systemic Sclerosis: Case Report.

Lung transplantation can be a life-saving measure for people with end-stage lung disease from systemic sclerosis. However, outcomes of lung transplantation may be compromised by gastrointestinal manifestations of systemic sclerosis, which can involve any part of the gastrointestinal tract. Esophageal and gastric disease can be managed by enteral feeding with the use of a gastrojejunal feeding tube. In this report, we describe the clinical courses of 2 lung transplant recipients with systemic sclerosis who experienced severe and prolonged barium-impaction ileus after insertion of a percutaneous gastrojejunal feeding tube.

Enchytraeid abundance in Araucaria Mixed Forest determined by cold and hot wet extraction.

Enchytraeids are small oligochaetes found worldwide in soils with sufficient moisture and organic matter, but scarcely studied in the Southern hemisphere. This is the third study on enchytraeid abundance in Brazil using wet extraction and the first carried out in Araucaria Mixed Forest (subtropical region). The sampling and extraction were based on the standard method ISO 23611-3/2007 using an adapted split soil corer and wet extraction with and without heat to assess the abundance of enchytraeids in a forest fragment at Embrapa Forestry in Colombo, Paraná State. The samplings were performed in 3 occasions between September 2011 and April 2012. The average numbers estimated by each method varied from appr. 2.000-12.000 (cold) and 5.000-12.000 ind./ m2 (hot), respectively, with a maximum of 44.000 ind./ m2 in one of the samples, the highest value reported so far in Brazil. The hot extraction was more advantageous, given the speed and preservation of the specimens in vivo, allowing taxonomic identification. Advantages and disadvantages of wet extractions compared to handsorting and formol methods are also discussed. Guaranidrilus, Hemienchytraeus, Enchytraeus, Fridericia and Achaeta were the genera identified in the samples.

Enchytraeid abundance in Araucaria Mixed Forest determined by cold and hot wet extraction

Abstract Enchytraeids are small oligochaetes found worldwide in soils with sufficient moisture and organic matter, but scarcely studied in the Southern hemisphere. This is the third study on enchytraeid abundance in Brazil using wet extraction and the first carried out in Araucaria Mixed Forest (subtropical region). The sampling and extraction were based on the standard method ISO 23611-3/2007 using an adapted split soil corer and wet extraction with and without heat to assess the abundance of enchytraeids in a forest fragment at Embrapa Forestry in Colombo, Paraná State. The samplings were performed in 3 occasions between September 2011 and April 2012. The average numbers estimated by each method varied from appr. 2.000-12.000 (cold) and 5.000-12.000 ind./ m2 (hot), respectively, with a maximum of 44.000 ind./ m2 in one of the samples, the highest value reported so far in Brazil. The hot extraction was more advantageous, given the speed and preservation of the specimens in vivo, allowing taxonomic identification. Advantages and disadvantages of wet extractions compared to handsorting and formol methods are also discussed. Guaranidrilus, Hemienchytraeus, Enchytraeus, Fridericia and Achaeta were the genera identified in the samples.

Resumo Os enquitreídeos são pequenos oligoquetas encontrados no mundo todo em solos com suficiente umidade e matéria orgânica, porém muito pouco estudados no hemisfério Sul. Este é o terceiro estudo sobre a abundância de enquitreídeos no Brasil utilizando o método de extração úmida e o primeiro realizado em Floresta Ombrófila Mista (região subtropical). A amostragem e extração foram baseadas no método padrão ISO 23611-3/2007, utilizando-se um trado desmontável adaptado e extração úmida com e sem aquecimento para acessar a abundância de enquitreídeo em um fragmento de floresta na Embrapa Florestas em Colombo, Paraná. As amostragens foram realizadas em três ocasiões entre setembro, 2011 e abril 2012. Os números médios estimados através de cada método variaram de 2.000-12.000 (frio) e 5.000-12.000 ind./ m2 (quente), respectivamente, e o máximo de 44.000 ind./ m2 em uma das amostras, o mais alto já relatado no Brasil. A extração quente foi a mais vantajosa, considerando a rapidez e preservação dos exemplares in vivo. As vantagens e desvantagens das extrações úmidas comparadas aos métodos de triagem manual e extração com formol foram discutidas. Os gêneros Guaranidrilus, Hemienchytraeus, Enchytraeus, Fridericia e Achaeta foram identificados nas amostras.

Enchytraeid abundance in Araucaria Mixed Forest determined by cold and hot wet extraction / Abundância de enquitreídeos na Floresta Ombrófila Mista determinada por extração úmida quente e fria

Abstract Enchytraeids are small oligochaetes found worldwide in soils with sufficient moisture and organic matter, but scarcely studied in the Southern hemisphere. This is the third study on enchytraeid abundance in Brazil using wet extraction and the first carried out in Araucaria Mixed Forest (subtropical region). The sampling and extraction were based on the standard method ISO 23611-3/2007 using an adapted split soil corer and wet extraction with and without heat to assess the abundance of enchytraeids in a forest fragment at Embrapa Forestry in Colombo, Paraná State. The samplings were performed in 3 occasions between September 2011 and April 2012. The average numbers estimated by each method varied from appr. 2.000-12.000 (cold) and 5.000-12.000 ind./ m2 (hot), respectively, with a maximum of 44.000 ind./ m2 in one of the samples, the highest value reported so far in Brazil. The hot extraction was more advantageous, given the speed and preservation of the specimens in vivo, allowing taxonomic identification. Advantages and disadvantages of wet extractions compared to handsorting and formol methods are also discussed. Guaranidrilus, Hemienchytraeus, Enchytraeus, Fridericia and Achaeta were the genera identified in the samples.

Resumo Os enquitreídeos são pequenos oligoquetas encontrados no mundo todo em solos com suficiente umidade e matéria orgânica, porém muito pouco estudados no hemisfério Sul. Este é o terceiro estudo sobre a abundância de enquitreídeos no Brasil utilizando o método de extração úmida e o primeiro realizado em Floresta Ombrófila Mista (região subtropical). A amostragem e extração foram baseadas no método padrão ISO 23611-3/2007, utilizando-se um trado desmontável adaptado e extração úmida com e sem aquecimento para acessar a abundância de enquitreídeo em um fragmento de floresta na Embrapa Florestas em Colombo, Paraná. As amostragens foram realizadas em três ocasiões entre setembro, 2011 e abril 2012. Os números médios estimados através de cada método variaram de 2.000-12.000 (frio) e 5.000-12.000 ind./ m2 (quente), respectivamente, e o máximo de 44.000 ind./ m2 em uma das amostras, o mais alto já relatado no Brasil. A extração quente foi a mais vantajosa, considerando a rapidez e preservação dos exemplares in vivo. As vantagens e desvantagens das extrações úmidas comparadas aos métodos de triagem manual e extração com formol foram discutidas. Os gêneros Guaranidrilus, Hemienchytraeus, Enchytraeus, Fridericia e Achaeta foram identificados nas amostras.

Quantitative trait loci mapping and gene network analysis implicate protocadherin-15 as a determinant of brain serotonin transporter expression.

Presynaptic serotonin (5-hydroxytryptamine, 5-HT) transporters (SERT) regulate 5-HT signaling via antidepressant-sensitive clearance of released neurotransmitter. Polymorphisms in the human SERT gene (SLC6A4) have been linked to risk for multiple neuropsychiatric disorders, including depression, obsessive-compulsive disorder and autism. Using BXD recombinant inbred mice, a genetic reference population that can support the discovery of novel determinants of complex traits, merging collective trait assessments with bioinformatics approaches, we examine phenotypic and molecular networks associated with SERT gene and protein expression. Correlational analyses revealed a network of genes that significantly associated with SERT mRNA levels. We quantified SERT protein expression levels and identified region- and gender-specific quantitative trait loci (QTLs), one of which associated with male midbrain SERT protein expression, centered on the protocadherin-15 gene (Pcdh15), overlapped with a QTL for midbrain 5-HT levels. Pcdh15 was also the only QTL-associated gene whose midbrain mRNA expression significantly associated with both SERT protein and 5-HT traits, suggesting an unrecognized role of the cell adhesion protein in the development or function of 5-HT neurons. To test this hypothesis, we assessed SERT protein and 5-HT traits in the Pcdh15 functional null line (Pcdh15(av-) (3J) ), studies that revealed a strong, negative influence of Pcdh15 on these phenotypes. Together, our findings illustrate the power of multidimensional profiling of recombinant inbred lines in the analysis of molecular networks that support synaptic signaling, and that, as in the case of Pcdh15, can reveal novel relationships that may underlie risk for mental illness.

Evaluation of heritable determinants of blood and brain serotonin homeostasis using recombinant inbred mice.

The biogenic amine serotonin (5-HT, 5-hydroxytryptamine) exerts powerful, modulatory control over multiple physiological functions in the brain and periphery, ranging from mood and appetite to vasoconstriction and gastrointestinal motility. In order to gain insight into shared and distinct molecular and phenotypic networks linked to variations in 5-HT homeostasis, we capitalized on the stable genetic variation present in recombinant inbred mouse strains. This family of strains, all derived from crosses between C57BL/6J and DBA/2J (BXD) parents, represents a unique, community resource with approximately 40 years of assembled phenotype data that can be exploited to explore and test causal relationships in silico. We determined levels of 5-HT and 5-hydroxyindoleacetic acid from whole blood, midbrain and thalamus/hypothalamus (diencephalon) of 38 BXD lines and both sexes. All 5-HT measures proved highly heritable in each region, although both gender and region significantly impacted between-strain correlations. Our studies identified both expected and novel biochemical, anatomical and behavioral phenotypes linked to 5-HT traits, as well as distinct quantitative trait loci. Analyses of these loci nominate a group of genes likely to contribute to gender- and region-specific capacities for 5-HT signaling. Analysis of midbrain mRNA variations across strains revealed overlapping gene expression networks linked to 5-HT synthesis and metabolism. Altogether, our studies provide a rich profile of genomic, molecular and phenotypic networks that can be queried for novel relationships contributing risk for disorders linked to perturbed 5-HT signaling.

Prediction of clinically relevant safety signals of nephrotoxicity through plasma metabolite profiling.

Addressing safety concerns such as drug-induced kidney injury (DIKI) early in the drug pharmaceutical development process ensures both patient safety and efficient clinical development. We describe a unique adjunct to standard safety assessment wherein the metabolite profile of treated animals is compared with the MetaMap Tox metabolomics database in order to predict the potential for a wide variety of adverse events, including DIKI. To examine this approach, a study of five compounds (phenytoin, cyclosporin A, doxorubicin, captopril, and lisinopril) was initiated by the Technology Evaluation Consortium under the auspices of the Drug Safety Executive Council (DSEC). The metabolite profiles for rats treated with these compounds matched established reference patterns in the MetaMap Tox metabolomics database indicative of each compound's well-described clinical toxicities. For example, the DIKI associated with cyclosporine A and doxorubicin was correctly predicted by metabolite profiling, while no evidence for DIKI was found for phenytoin, consistent with its clinical picture. In some cases the clinical toxicity (hepatotoxicity), not generally seen in animal studies, was detected with MetaMap Tox. Thus metabolite profiling coupled with the MetaMap Tox metabolomics database offers a unique and powerful approach for augmenting safety assessment and avoiding clinical adverse events such as DIKI.

Case-control study of a multistate equine herpesvirus myeloencephalopathy outbreak.

BACKGROUND: A large multistate outbreak of equine herpesvirus myeloencephalopathy (EHM) occurred in May 2011 among horses that participated in a competitive event. OBJECTIVE: To identify EHM risk factors among horses with a common exposure venue. ANIMALS: A total of 123 horses: 19 horses with EHM, 14 equine herpesvirus-1 cases with no reported neurologic signs, and 90 control horses. METHODS: EHM case survey data were compared with data from EHV-1 cases with no neurologic signs and healthy controls using univariable and multivariable methods. RESULTS: Significant factors associated with higher risk for EHM compared with EHV-1 cases with no neurologic signs were (1) greater number of biosecurity risks at the event, (2) female sex, (3) increasing number of classes competed in at the event, and (4) an interaction between sex and number of classes competed in. In the EHM versus controls comparison, in addition to sex and biosecurity risks, factors associated with higher EHM risk included EHV-1 vaccination in the 5 weeks before the event and increasing number of events attended in April 2011; zinc dietary supplementation was associated with decreased risk. An interaction between sex and the number of events attended in April 2011 also was significant. CONCLUSIONS AND CLINICAL IMPORTANCE: Findings from this study suggest that dietary zinc supplementation may be associated with decreased risk of EHM. Several factors were associated with increased risk of EHM. Additional investigations of factors associated with risk of EHM are warranted to evaluate the importance of these factors in this complex disease of horses.

First Report of Puccinia emaculata Infection on Switchgrass in Virginia.

During the spring of 2007, approximately 2,000 switchgrass plants, representing 168 core switchgrass germplasm, were established in a field nursery at the Virginia Tech Kentland Farm Research Center in Montgomery County, VA. These germplasms were originally obtained from the USDA Plant Genetic Resources Conservation Unit (Griffin, GA) and included both lowland and upland ecotypes. After planting, the switchgrass plants were allowed to establish for 3 years. In the summers of 2009, 2010, and 2011, a substantial infection of a foliar rust pathogen was observed on more than 90% of switchgrass lines in the field. The infected leaves had dark brown pustules that were arranged in a linear fashion between the veins and mainly located on the adaxial surface of the leaf. We observed the infected leaf samples under a microscope and isolated the urediniospores. The urediniospores were brown, round, and averaged 26.08 ± 1.67 µm long and 24.65 ± 1.66 µm wide. Teliospores were observed late in the summers of 2009 and 2010. The teliospores were two-celled, oblong to ellipsoid in shape, and averaged 32.23 ± 3.07 µm in length. The apical cell width averaged 17.6 ± 1.83 µm and the basal cell width averaged 15.08 ± 1.75 µm. The morphology of both the urediniospores and teliospores were similar to Puccinia emaculata Schw. (P. emaculata) (2,3). Using previously established rust disease scoring methods (1), the infected switchgrass lines were evaluated and scored for infection severity on a 0 to 9 scale based on the percentage of leaf area infected, with 0 being either highly resistant or escaping infection and 9 being highly susceptible. The majority of upland switchgrass cultivars, including Caddo, Cave-in-rock, Blackwell, Sunburst, Pathfinder, and Dacotah, were moderately to highly susceptible to this rust pathogen and on average, scored between 7 and 8. However, the majority of lowland switchgrass cultivars, including Alamo, Kanlow, TEM-SEC, TEM-SLC, and TEM-LoDorm, were moderately to highly resistant and scored on average between 3 and 4. This result is consistent with previous reports that show that lowland ecotypes are more resistant to rust diseases (1). To further validate the identity of the rust pathogen, we designed two primers (5'-CCAGTAACGGCGAGTGAAGAG-3' and 5'-CGACTTCCATGGCCACCGTGCGGCTGTCT-3') based on the 18S rDNA sequence of P. emaculata (3). DNA was extracted from bulk infected leaf material for PCR amplification. The 1.2 kb PCR product was isolated and sent for DNA sequencing. The DNA sequence was 98% identical to the 18S rDNA sequence of P. emaculata (EU915294.1). To our knowledge, this study represents the first report of rust on a wide range of various switchgrass cultivars in Virginia. References: (1) D. M. Gustafson et al. Crop Sci. 43:755, 2003. (2) R. L. Hirsch et al. Plant Dis. 94:381, 2010. (3) J. Zale et al. Plant Dis. 92:1710, 2008.

Spinach latent virus Infecting Tomato in Virginia, United States.

Plants in a single field of commercial tomato (Solanum lycopersicum) of unidentified cultivars in Virginia in July, 2012, were observed showing stunting, leaf distortion, twisting and thickening, discoloration, and color streaking and ringspots on fruits. Serological tests were negative for Cucumber mosaic virus, Groundnut ringspot virus, Tomato spotted wilt virus, Tomato chlorotic spot virus, Impatiens necrotic spot virus, Tobacco mosaic virus, and Tomato bushy stunt virus (Agdia, Inc., Elkhart, IN). Using a membrane-based macroarray (3), hybridization was observed to 8 of 9 70-mer oligonucleotide probes of Spinach latent virus (SpLV; genus Ilarvirus, family Bromoviridae). To confirm the hybridization results, complementary DNA (cDNA) was synthesized using random hexamers and MMLV reverse transcriptase (Promega, Madison, WI), followed by PCR amplification using ilarvirus degenerate primers (4). Fragments of approximately 380 bp were amplified and directly sequenced (GenBank Accession KC_466090); a BLAST search showed a 99% identity to the SpLV RNA 2 reference genome (NC_003809). Primers for SpLV RNA1 (SpLVRNA1f-GGTGTCACCATGCAAACTGG, SpLVRNA1r-AGCTCTTCGTAATAGGCCTGC) and SpLV RNA3 (SpLVCPf-GAAGTCTTTCCCAGGTGAGCA, SpLVCPr-AGGTGGGCATATGGACTTGG) were designed and cDNA was amplified using the IQ supermix (Biorad, Hercules, CA) with thermocycling of 94°C for 4 min, 35× (94°C 45 s, 55°C 45 s, 72°C 45 s), and 72°C for 10 min. The resulting fragments of 538 bp for RNA1 (KC_466088) and 661 bp for RNA3 (KC_466089) showed 100% identity to reference genome sequences for SpLV (NC_003808 and NC_003810, respectively). To demonstrate virus transmissibility, Chenopodium quinoa plants were mechanically inoculated using tomato leaf material (same source described above) ground in 30 mM Na2HPO4 buffer, pH 7.0. Necrotic spots developed on the inoculated leaves 10 dpi. Younger, non-inoculated leaves showed yellow mottling and tested positive for SpLV by RT-PCR (two of two plants tested). The detection of SpLV is rarely reported, with only one record from the United States (2). Although SpLV is described as a latent virus, it has been found associated with tomato fruit symptoms in New Zealand (1). It is not known if the fruit ringspot and other symptoms on the Virginia samples were due to virus infection. Since SpLV is seed-transmissible and seed production takes place in different parts of the world, it has the potential to spread with germplasm and become more widespread in North America. References: (1) B. S. M. Lebas et al. Plant Dis. 91:228, 2007. (2) H. Y. Liu and J. E. Duffus. Phytopathology 76:1087, 1986. (3) K. L. Perry and X. Lu. Phytopathology 100:S100, 2010. (4) M. Untiveros, et al. J. Virol. Methods 165:97, 2010.

Salmonella prevalence and antimicrobial susceptibility from the National Animal Health Monitoring System Swine 2000 and 2006 studies.

Concern about Salmonella contamination of food is compounded by fear that antimicrobials traditionally used to combat the infection will become useless due to rising antibiotic resistance. Livestock, in particular swine, often are blamed for illnesses caused by Salmonella and for increasing antibiotic resistance due to use of antibiotics in pigs. As part of the National Animal Health Monitoring System Swine 2000 and 2006 studies, swine fecal samples were cultured for Salmonella. These samples were collected from 123 operations in 17 states in 2000 and from 135 operations in 17 states in 2006. At each operation, 50 and 60 fecal samples were collected from late finisher pig pens in 2000 and 2006, respectively. Salmonella isolates were characterized to determine serogroup and serotype and were tested for susceptibility to a panel of 17 and 15 antimicrobial drugs in 2000 and 2006, respectively. A total of 5,470 and 7,788 samples were cultured for Salmonella in 2000 and 2006, respectively. Overall, 6.2% of the samples and 34.2% of the farms were positive for Salmonella in 2000. In 2006, 7.2% of the samples and 52.6% of the farms were positive. Salmonella Derby, Salmonella Typhimurium var. 5- (formerly Salmonella Typhimurium var. Copenhagen), and Salmonella Agona were the three serotypes most often recovered in both study years. The most common antimicrobial resistance pattern for Salmonella Derby in the two study years was resistance to streptomycin, sulfisoxazole, and tetracycline. Most isolates were resistant to tetracycline, sulfisoxazole, and streptomycin in both study years. The proportion of Salmonella isolates that were susceptible to all antimicrobials (pansusceptible) was 38.1% in 2000 and 20.4% in 2006. The proportion of Salmonella isolates that were resistant to three or more antimicrobials (multidrug resistant) was similar in 2000 and in 2006 (52.8 and 57.7%, respectively).

First Report of Bacterial Spot of Peony Caused by a Xanthomonas sp. in the United States.

In early May 2008 and 2009, peony samples (Paeonia spp.) with symptoms of leaf spot and blight were submitted to the Virginia Tech Plant Disease Clinic. The 2008 peony was an unknown cultivar from a northern Virginia landscape. The three cultivars (Dr. Alexander Fleming, Felix Crousse, and Karl Rosenfield) submitted in 2009 were from a commercial nursery in southwestern Virginia that was reporting leaf spot progressing to severe blight, which rendered plants unsalable, on 75% of a 1,219 m2 block during a 10-day period of heavy rainfall. Bacterial streaming from spots was observed. On the basis of phenotypic and biochemical tests, the isolates were determined to be xanthomonads. Two isolates (one recovered from the 2008 sample and one from the 2009 sample) were used in the following work. Isolates were characterized by multilocus sequencing (MLST) (4). PCR reactions were prepared and cycled using 2X ImmoMix (Bioline, Tauton, MA) according to manufacturer's recommendations with an annealing temperature of 58°C. Template DNA was added by touching a single colony with a 20-µl pipette tip and placing the tip into the reaction mix for 1 min. Four bands of the expected size were visualized on an electrophoresis gel and cleaned products were sequenced in forward and reverse directions at the University of Chicago, Cancer Research Center DNA Sequencing Facility. Corresponding gene fragments of each isolate were identical. A consensus sequence (PAMDB Isolate ID No. 936) for each of the four gene fragments was constructed and compared with sequences in NCBI ( http://www.ncbi.nlm.nih.gov/nuccore/ ) and PAMDB ( http://genome.ppws.vt.edu/cgi-bin/MLST/home.pl ) (1) databases using Blastn (2). No perfect match was found. Genetic distances between the peony isolates and all strains in PAMDB were determined by MegAlign (Lasergene; DNAStar, Madison, WI). The Xanthomonas strain most similar to the isolates recovered from the peony samples was Xanthomonas hortorum pv. hederae ICMP 1661 with a genetic distance of 0.023; this strongly suggests that the peony isolates belong to X. hortorum. For Koch's postulates, six surface-disinfested young leaflets from Paeonia lactiflora 'Karl Rosenfield' were inoculated by forcefully spraying a phosphate-buffered saline suspension of each bacterial isolate (~4.3 × 109 CFU/ml) into the underside of the leaf until leaf tissue appeared water soaked. Controls were inoculated similarly with phosphate-buffered saline solution. Moist chambers with inoculated leaves were incubated at ambient temperature under two 48W fluorescent grow lights with 12 h of light and dark. Circular spots were observed on leaves inoculated with the 2009 and 2008 isolates in 18 and 20 days, respectively. No symptoms were observed on controls. Bacterial streaming from leaf spots was observed by phase-contrast microscopy; bacteria were isolated and confirmed to be identical to the original isolates by the methods described above. To our knowledge, this is the first report of a Xanthomonas sp. causing leaf spot and blight on peony. Although bacterial blight of peony has been attributed to a xanthomonad in recent years, the pathogen had not been further characterized (3). References: (1) N. F. Almeida et al. Phytopathology 100:208, 2010. (2) D. J. Altschul et al. J. Mol. Biol. 215:403, 1990. (3) M. L. Gleason et al. Diseases of Herbaceous Perennials. The American Phytopathological Society, St. Paul, MN. 2009. (4) J. M. Young et al. Syst. Appl. Microbiol. 31:366, 2008.

Elevated 5-HT 2A receptors in postmortem prefrontal cortex in major depression is associated with reduced activity of protein kinase A.

Previous human postmortem brain tissue research has implicated abnormalities of 5-HT receptor availability in depression and suicide. Although altered abundance of 5-HT 1A, 5-HT 2A, and 5-HT 2C receptors (5-HT(1A), 5-HT(2A), and 5-HT(2C)) has been reported, the causes remain obscure. This study evaluated the availability of these three receptor subtypes in postmortem brain tissue specimens from persons with a history of major depression (MDD) and normal controls and tested the relationships to protein kinases A and C (PKA, PKC). Samples were obtained from postmortem brain tissue (Brodmann area 10) from 20 persons with a history of MDD and 20 matched controls as determined by a retrospective diagnostic evaluation obtained from family members. Levels of 5-HT(1A), 5-HT(2A), and 5-HT(2C) receptor were quantitated via Western blot analyses. Basal and stimulated PKA and PKC activity were also determined. The depressed samples showed significantly increased 5-HT(2A) receptor abundance relative to controls, but no differences in 5-HT(1A) or 5-HT(2C) receptors. Basal and cyclic AMP-stimulated PKA activity was also reduced in the depressed sample; PKC activity was not different between groups. 5-HT(2A) receptor availability was significantly inversely correlated with PKC activity in controls, but with PKA activity in the depressed sample. Increased 5-HT(2A) receptor abundance and decreased PKA activity in the depressed sample are consistent with prior reports. The correlation of 5-HT(2A) receptor levels with PKA activity in the depressed group suggests that abnormalities of 5-HT(2A) receptor abundance may depend on receptor uncoupling and heterologous regulation by PKA.

Chronic aspiration of gastric fluid induces the development of obliterative bronchiolitis in rat lung transplants.

Long-term survival of a pulmonary allograft is currently hampered by obliterative bronchiolitis (OB), a form of chronic rejection that is unique to lung transplantation. While tracheobronchial aspiration from gastroesophageal reflux disease (GERD) has clinically been associated with OB, no experimental model exists to investigate this problem. Using a WKY-to-F344 rat orthotopic left lung transplant model, the effects of chronic aspiration on pulmonary allograft were evaluated. Recipients received cyclosporine with or without 8 weekly aspirations of gastric fluid into the allograft. Six (66.7%) of 9 allografts with aspiration demonstrated bronchioles with surrounding monocytic infiltrates, fibrosis and loss of normal lumen anatomy, consistent with the development of OB. In contrast, none of the allografts without aspiration (n = 10) demonstrated these findings (p = 0.002). Of the grafts examined grossly, 83% of the allografts with chronic aspiration but only 20% without aspiration appeared consolidated (p = 0.013). Aspiration was associated with increased levels of IL-1 alpha, IL-1 beta, IL-6, IL-10, TNF-alpha and TGF-beta in BAL and of IL-1 alpha, IL-4 and GM-CSF in serum. This study provides experimental evidence linking chronic aspiration to the development of OB and suggests that strategies aimed at preventing aspiration-related injuries might improve outcomes in clinical lung transplantation.

Multivariate permutation analysis associates multiple polymorphisms with subphenotypes of major depression.

Unipolar major depressive disorder (MDD) is a prevalent, disabling condition with multiple genetic and environmental factors impacting disease risk. The diagnosis of MDD relies on a cumulative measure derived from multiple trait dimensions and alone is limited in elucidating MDD genetic determinants. We and others have proposed that MDD may be better dissected using paradigms that assess how specific genes associate with component features of MDD. This within-disease design requires both a well-phenotyped cohort and a robust statistical approach that retains power with multiple tests of genetic association. In the present study, common polymorphic variants of genes related to central monoaminergic and cholinergic pathways that previous studies align with functional change in vitro or depression associations in vivo were genotyped in 110 individuals with unipolar MDD. Subphenotypic characteristics were examined using responses to individual items assessed with the Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders (DSM IV), the 17-item Hamilton Rating Scale for Depression (HAM-D) and the NEO Five Factor Inventory. Multivariate Permutation Testing (MPT) was used to infer genotype-phenotype relationships underlying dimensional findings within clinical categories. MPT analyses show significant associations of the norepinephrine transporter (NET, SLC6A2) -182 T/C (rs2242446) with recurrent depression [odds ratio, OR = 4.15 (1.91-9.02)], NET -3081 A/T (rs28386840) with increase in appetite [OR = 3.58 (1.53-8.39)] and the presynaptic choline transporter (CHT, SLC5A7) Ile89Val (rs1013940) with HAM-D-17 total score {i.e. overall depression severity [OR = 2.74 (1.05-7.18)]}. These relationships illustrate an approach to the elucidation of gene influences on trait components of MDD and with replication, may help identify MDD subpopulations that can benefit from more targeted pharmacotherapy.

An evaluation of scrapie surveillance in the United States.

Animal health surveillance systems should reflect national disease control priorities and promote the best use of public resources by maximizing effectiveness and efficiency. A surveillance system should be routinely evaluated to assess the degree to which the system accomplishes these goals, fulfills its stated objectives, and meets accepted surveillance standards. In the United States, there are a number of disparate endemic disease surveillance and eradication programs. The National Animal Health Surveillance System is a federal initiative designed to combine animal health surveillance and monitoring activities into a comprehensive and coordinated system. A protocol has been developed to facilitate the evaluation of animal health surveillance systems and investigate opportunities for coordination between the different surveillance and eradication programs. The evaluation protocol was based largely on protocols developed for public health but adapted for the specific needs and goals of animal health surveillance. The evaluation process was designed to identify program strengths and areas for improvement and facilitate the system's adaptability to changing situations. The evaluation protocol was applied to the scrapie surveillance system in the United States; scrapie surveillance was found to be an important part of surveillance for transmissible spongiform encephalopathies. Results from the evaluation of sensitivity, sampling methods and representativeness are presented.