Intramammary lipopolysaccharide challenge in early- versus mid-lactation dairy cattle: Immune, production, and metabolic responses.

Study objectives were to compare the immune response, metabolism, and production following intramammary LPS (IMM LPS) administration in early and mid-lactation cows. Early (E-LPS; n = 11; 20 ± 4 DIM) and mid- (M-LPS; n = 10; 155 ± 40 DIM) lactation cows were enrolled in an experiment consisting of 2 periods (P). During P1 (5 d) cows were fed ad libitum and baseline data were collected, including liver and muscle biopsies. At the beginning of P2 (3 d) cows received 10 mL of sterile saline containing 10 µg of LPS from Escherichia coli O111:B4/mL into the left rear quarter of the mammary gland, and liver and muscle biopsies were collected at 12 h after LPS. Tissues were analyzed for metabolic flexibility, which measures substrate switching capacity from pyruvic acid to palmitic acid oxidation. Data were analyzed with the MIXED procedure in SAS 9.4. Rectal temperature was assessed hourly for the first 12 h after LPS and every 6 h thereafter for the remainder of P2. All cows developed a febrile response following LPS, but E-LPS had a more intense fever than M-LPS cows (0.7°C at 5 h after LPS). Blood samples were collected at 0, 3, 6, 9, 12, 24, 36, 48, and 72 h after LPS for analysis of systemic inflammation and metabolism parameters. Total serum Ca decreased after LPS (26% at 6 h nadir) but did not differ by lactation stage (LS). Circulating neutrophils decreased, then increased after LPS in both LS, but E-LPS had exaggerated neutrophilia (56% from 12 to 48 h) compared with M-LPS. Haptoglobin increased after LPS (15-fold) but did not differ by LS. Many circulating cytokines were increased after LPS, and IL-6, IL-10, TNF-α, MCP-1, and IP-10 were further augmented in E-LPS compared with M-LPS cows. Relative to P1, all cows had reduced milk yield (26%) and DMI (14%) on d 1 that did not differ by LS. Somatic cell score increased rapidly in response to LPS regardless of LS and gradually decreased from 18 h onwards. Milk component yields decreased after LPS. However, E-LPS had increased fat (11%) and tended to have increased lactose (8%) yield compared with M-LPS cows throughout P2. Circulating glucose was not affected by LPS. Nonesterified fatty acids (NEFA) decreased in E-LPS (29%) but not M-LPS cows. ß-Hydroxybutyrate slightly increased (14%) over time after LPS regardless of LS. Insulin increased after LPS in all cows, but E-LPS had blunted hyperinsulinemia (52%) compared with M-LPS cows. Blood urea nitrogen increased after LPS, and the relative change in BUN was elevated in E-LPS cows compared with M-LPS cows (36% and 13%, respectively, from 9 to 24 h). During P1, metabolic flexibility was increased in liver and muscle in early lactating cows compared with mid-lactation cows, but 12 h after LPS, metabolic flexibility was reduced and did not differ by LS. In conclusion, IMM LPS caused severe immune activation, and E-LPS cows had a more intense inflammatory response compared with M-LPS cows, but the effects on milk synthesis was similar between LS. Some parameters of the E-LPS metabolic profile suggest continuation of metabolic adjustments associated with early lactation to support both a robust immune system and milk synthesis.

Validation of an Oral Disease Severity Score (ODSS) tool for use in oral mucous membrane pemphigoid.

BACKGROUND: Mucous membrane pemphigoid (MMP) is a rare autoimmune bullous disease predominantly affecting the oral mucosa. Optimal management relies upon thorough clinical assessment and documentation at each visit. OBJECTIVES: The primary aim of this study was to validate the Oral Disease Severity Score (ODSS) for the assessment of oral involvement in MMP. We also compared its inter- and intraobserver reliability with those of the oral parts of the Mucous Membrane Pemphigoid Disease Area Index (MMPDAI), Autoimmune Bullous Skin Disorder Intensity Score (ABSIS) and Physician's Global Assessment (PGA). METHODS: Fifteen patients with mild-to-moderately severe oral MMP were scored for disease severity by 10 oral medicine clinicians from four U.K. centres using the ODSS, the oral sections of MMPDAI and ABSIS, and PGA. Two clinicians rescored all patients after 2 h. RESULTS: In terms of reliability, the interobserver ODSS total score intraclass correlation coefficient (ICC) was 0·97, MMPDAI activity 0·59 and damage 0·15, ABSIS total 0·84, and PGA 0·72. The intraobserver ICCs (two observers) for ODSS total were 0·97 and 0·93; for MMPDAI activity 0·93 and 0·70 and damage 0·93 and 0·79; for ABSIS total 0·99 and 0·94; and for PGA 0·92 and 0·94. Convergent validity between ODSS and MMPDAI was good (correlation coefficient 0·88). The mean ± SD time for completion of ODSS was 93 ± 31 s, with MMPDAI 102 ± 24 s and ABSIS involvement 71 ± 18 s. The PGA took < 5 s. CONCLUSIONS: This study has validated the ODSS for the assessment of oral MMP. It has shown superior interobserver agreement over MMPDAI, ABSIS and PGA, and superior intraobserver reliability to MMPDAI. It is quick and easy to perform. What's already known about this topic? There are no validated scoring methodologies for oral mucous membrane pemphigoid (MMP). Proposed disease activity scoring tools for MMP include the Mucous Membrane Disease Area Index (MMPDAI) and the Autoimmune Bullous Skin Disorder Intensity Score (ABSIS). The Oral Disease Severity Score (ODSS) has been validated for use in oral pemphigus vulgaris (PV). It has been shown to be reliable and sensitive in both lichen planus (LP) and MMP. What does this study add? The ODSS has been shown to be a thorough, sensitive and reproducible, yet quick scoring tool for the assessment of oral involvement in MMP. Its versatility for use in oral PV, MMP and LP is an added advantage over other scoring methodologies. What are the clinical implications of this work? We propose that the ODSS be used as a clinical scoring tool for monitoring activity in oral MMP in clinical practice as well as for use in multicentre studies.

Indexing sensory plasticity: Evidence for distinct Predictive Coding and Hebbian learning mechanisms in the cerebral cortex.

The Roving Mismatch Negativity (MMN), and Visual LTP paradigms are widely used as independent measures of sensory plasticity. However, the paradigms are built upon fundamentally different (and seemingly opposing) models of perceptual learning; namely, Predictive Coding (MMN) and Hebbian plasticity (LTP). The aim of the current study was to compare the generative mechanisms of the MMN and visual LTP, therefore assessing whether Predictive Coding and Hebbian mechanisms co-occur in the brain. Forty participants were presented with both paradigms during EEG recording. Consistent with Predictive Coding and Hebbian predictions, Dynamic Causal Modelling revealed that the generation of the MMN modulates forward and backward connections in the underlying network, while visual LTP only modulates forward connections. These results suggest that both Predictive Coding and Hebbian mechanisms are utilized by the brain under different task demands. This therefore indicates that both tasks provide unique insight into plasticity mechanisms, which has important implications for future studies of aberrant plasticity in clinical populations.

Neural plasticity is modified over the human menstrual cycle: Combined insight from sensory evoked potential LTP and repetition suppression.

In healthy women, fluctuations in hormones including progesterone and oestradiol lead to functional changes in the brain over the course of each menstrual cycle. Though considerable attention has been directed towards understanding changes in human cognition over the menstrual cycle, changes in underlying processes such as neural plasticity have largely only been studied in animals. In this study we explored predictive coding and repetition suppression via the roving mismatch negativity paradigm as a model of short-term plasticity (Garrido, Kilner, Kiebel, et al., 2009), and Hebbian learning via visual sensory long-term potentiation (LTP) as a model of long-term plasticity (Teyler et al., 2005). Electroencephalography (EEG) was recorded in 20 females during their early follicular and mid-luteal phases. Event-related potential (ERP) analyses were complemented with dynamic causal modelling (DCM) to characterise changes in the underlying neural architecture. More sustained variability in the ERP response to a change in tone during the luteal phase are interpreted as a delayed habituation of the P3a component in the luteal relative to the follicular phase. The additional increased forward connection strength over tone repetitions compared to the follicular phase suggests that, in this phase, females may be less efficient when processing deviations from predicted sensory input (error). In contrast, there appears to be no reliable change in sensory LTP. This suggests that predictive coding, but not Hebbian plasticity is modified in the mid-luteal compared to the follicular phase, at least at the days of the menstrual cycle tested. This finding implicates the human menstrual cycle in complex changes in neural plasticity and provides further evidence for the importance of considering the menstrual cycle when including females in electrophysiological research.

An Oral Disease Severity Score validated for use in oral pemphigus vulgaris.

BACKGROUND: Pemphigus vulgaris (PV) is a rare autoimmune bullous disease, which can present with recalcitrant oral mucosal lesions. Optimal management of PV relies upon careful clinical assessment and documentation. OBJECTIVES: The primary aim of this study was to validate the Oral Disease Severity Score (ODSS) for the assessment of oral involvement in PV. A secondary aim was to compare its inter- and intraobserver variability and ease of use with the Physician's Global Assessment (PGA) and the oral scoring methods used in the Autoimmune Bullous Skin Disorder Intensity Score (ABSIS) and the Pemphigus Disease Area Index (PDAI). METHODS: Fifteen patients with mild-to-moderately severe oral PV were scored for disease severity by 10 oral medicine clinicians using the ODSS, the PGA and the oral sections of ABSIS and PDAI. Two clinicians rescored all patients after a minimum 2-h interval. RESULTS: Interobserver reliability was assessed using an intraclass correlation coefficient (ICC). For the ODSS total score the ICC was 0·83, for PDAI (oral total activity) 0·79, ABSIS (oral total) 0·71 and PGA 0·7. Intraobserver agreement between initial scoring and rescoring of the same patient by two clinicians demonstrated an ICC for each of 0·97 and 0·96 for ODSS total score; 0·99 and 0·82 for PDAI oral activity; 0·86 and 0·45 for ABSIS total; and 0·99 and 0·64 for PGA. Convergent validity was good, with a correlation coefficient > 0·5 (P < 0·001). The mean ± SD times taken to complete each scoring method were ODSS 76 ± 37 s, PDAI 117 ± 16 s and ABSIS 75 ± 19 s. CONCLUSIONS: This study has validated the ODSS for the assessment of oral PV. It has shown superior inter- and intraobserver reliability to PDAI, ABSIS and PGA and is quick to perform.

Fiber hypertrophy and increased oxidative capacity can occur simultaneously in pig glycolytic skeletal muscle.

An inverse relationship between skeletal muscle fiber cross-sectional area (CSA) and oxidative capacity suggests that muscle fibers hypertrophy at the expense of oxidative capacity. Therefore, our objective was to utilize pigs possessing mutations associated with increased oxidative capacity [AMP-activated protein kinase (AMPKγ3(R200Q))] or fiber hypertrophy [ryanodine receptor 1 (RyR1(R615C))] to determine if these events occur in parallel. Longissimus muscle was collected from wild-type (control), AMPKγ3(R200Q), RyR1(R615C), and AMPKγ3(R200Q)-RyR1(R615C) pigs. Regardless of AMPK genotype, RyR(R615C) increased fiber CSA by 35%. In contrast, AMPKγ3(R200Q) pig muscle exhibited greater citrate synthase and ß-hydroxyacyl CoA dehydrogenase activity. Isolated mitochondria from AMPKγ3(R200Q) muscle had greater maximal, ADP-stimulated oxygen consumption rate. Additionally, AMPKγ3(R200Q) muscle contained more (∼50%) of the mitochondrial proteins succinate dehydrogenase and cytochrome c oxidase and more mitochondrial DNA. Surprisingly, RyR1(R615C) increased mitochondrial proteins and DNA, but this was not associated with improved oxidative capacity, suggesting that altered energy metabolism in RyR1(R615C) muscle influences mitochondrial proliferation and protein turnover. Thus pigs that possess both AMPKγ3(R200Q) and RyR(R615C) exhibit increased muscle fiber CSA as well as greater oxidative capacity. Together, our findings support the notion that hypertrophy and enhanced oxidative capacity can occur simultaneously in skeletal muscle and suggest that the signaling mechanisms controlling these events are independently regulated.

Chickens from lines selected for high and low body weight show differences in fatty acid oxidation efficiency and metabolic flexibility in skeletal muscle and white adipose tissue.

OBJECTIVE: The Virginia lines of chickens have resulted from more than 55 generations of artificial selection for low (LWS) or high (HWS) juvenile body weight. We hypothesized that the relative hyperphagia and greater body weight in juvenile HWS chickens are associated with altered fatty acid oxidation efficiency and metabolic flexibility in tissues associated with energy sensing and storage, and relative cellular hypertrophy in white adipose tissue. METHODS: Hypothalamus, liver, pectoralis major, gastrocnemius, abdominal fat, clavicular fat and subcutaneous fat were collected from the juvenile (56-65 days old) LWS and HWS chickens for metabolic, gene expression and histological assays. RESULTS: The HWS chickens had reduced fatty acid oxidation efficiency in abdominal fat (P<0.0001) and reduced rates of oxidation in abdominal fat and gastrocnemius (P<0.0001) as compared with the LWS. There was reduced citrate synthase activity in white adipose tissue (P<0.0001) and greater metabolic inflexibility in skeletal muscle (P=0.006) of the HWS compared with the LWS. Greater pyruvate dehydrogenase kinase 4 (PDK4) and forkhead box O1A (FoxO1) mRNA were found in skeletal muscle and white adipose tissue of 56-day-old HWS than LWS. Expression of peroxisome proliferator-activated receptor γ (PPARγ) in all adipose tissue depots was greater (P<0.05) in LWS than in HWS chickens. The HWS chickens had larger (P<0.0001) and fewer (P<0.0001) adipocytes per unit area than the LWS. CONCLUSION: Compared with the LWS, the HWS chickens have impaired metabolic flexibility and fatty acid oxidation efficiency due to greater pyruvate dehydrogenase activity to accommodate the influx of acetyl-CoA from fatty acid oxidation in skeletal muscle and adipose tissue. These metabolic adaptations can be linked to differences in gene expression regulation, adipocyte cellularity and body composition between the lines, which may provide valuable insight into metabolic disorders in other species.

Prolonged voluntary wheel running reveals unique adaptations in mdx mice treated with microdystrophin constructs ± the nNOS-binding site.

We tested the effects of prolonged voluntary wheel running on the muscle function of mdx mice treated with one of two different microdystrophin constructs. At 7 weeks of age mdx mice were injected with a single dose of AAV9-CK8-microdystrophin with (gene therapy 1, GT1) or without (gene therapy 2, GT2) the nNOS-binding domain and were assigned to one of four gene therapy treated groups: mdxRGT1 (run, GT1), mdxGT1 (no run, GT1), or mdxRGT2 (run,GT2), mdxGT2 (no run, GT2). There were two mdx untreated groups injected with excipient: mdxR (run, no gene therapy) and mdx (no run, no gene therapy). A third no treatment group, Wildtype (WT) received no injection and did not run. mdxRGT1, mdxRGT2 and mdxR performed voluntary wheel running for 52 weeks; WT and remaining mdx groups were cage active. Robust expression of microdystrophin occurred in diaphragm, quadriceps, and heart muscles of all treated mice. Dystrophic muscle pathology was high in diaphragms of non-treated mdx and mdxR mice and improved in all treated groups. Endurance capacity was rescued by both voluntary wheel running and gene therapy alone, but their combination was most beneficial. All treated groups increased in vivo plantarflexor torque over both mdx and mdxR mice. mdx and mdxR mice displayed ∼3-fold lower diaphragm force and power compared to WT values. Treated groups demonstrated partial improvements in diaphragm force and power, with mdxRGT2 mice experiencing the greatest improvement at ∼60% of WT values. Evaluation of oxidative red quadriceps fibers revealed the greatest improvements in mitochondrial respiration in mdxRGT1 mice, reaching WT levels. Interestingly, mdxGT2 mice displayed diaphragm mitochondrial respiration values similar to WT but mdxRGT2 animals showed relative decreases compared to the no run group. Collectively, these data demonstrate that either microdystrophin construct combined with voluntary wheel running increased in vivo maximal muscle strength, power, and endurance. However, these data also highlighted important differences between the two microdystrophin constructs. GT1, with the nNOS-binding site, improved more markers of exercise-driven adaptations in metabolic enzyme activity of limb muscles, while GT2, without the nNOS-binding site, demonstrated greater protection of diaphragm strength after chronic voluntary endurance exercise but decreased mitochondrial respiration in the context of running.

Staffs' knowledge and perceptions of working with women with intellectual disabilities and mental health problems.

AIM: There is a growing evidence of the physical and mental health inequalities in people with intellectual disability (ID) although less has been written concerning the mental health of women with ID (International Association for the Scientific Study of Intellectual Disabilities). This is compared with the substantive literature published within mainstream psychiatry on gender. The aim of this study was to explore a range of health and social care staffs' knowledge and perceptions of caring for women with ID who have mental health problems focusing upon risk and resilient/protective factors. METHOD: A qualitative methodology was used. Eight focus groups were conducted with hospital, community and residential staff across one region of the UK. The focus groups were audiotaped and the transcriptions were subjected to a thematic content analysis using Newell & Burnard's framework. FINDINGS: Six inter-related risk factors were identified by the participants as potential causes for the women with ID to develop a mental illness and these were: having an ID and being female, unmet expectations, dysfunctional family upbringing, unstable relationships/loss of children, domestic violence and negative life experiences. Few of the participants acknowledged hormonal issues as a risk factor. Resilient/protective factors included being proactive, greater community participation, early recognition and mental health maintenance. CONCLUSION: These results are discussed in light of current developments and policy within mainstream psychiatric gender approaches. Greater recognition of a proactive health approach for both staff and women with an ID is recommended.

Antibody-dependent lymphocytotoxicity induced by immunoglobulin G from Hodgkin's disease splenic lymphocytes.

Immunoglobulin G, produced in cultures of splenic lymphocytes obtained from patients with Hodgkin's disease, bound to a population of homologous peripheral blood lymphocytes and initiated antibody-dependent cell cytotoxicity in cultures from five out of eight patients. Two patients whose cultures produced negative results had minimal disease; the other was in remission. The target cells appear to be T lymphocytes; the effector cells bear Fc receptors that are inhibited by antigen-antibody complexes. Antibody-dependent cell cytotoxicity events may produce the anergy and lymphopenia often seen in Hodgkin's disease.

Imaging photopolarimeter on pioneer saturn.

An imaging photopolarimeter aboard Pioneer 11, including a 2.5-centimeter telescope, was used for 2 weeks continuously in August and September 1979 for imaging, photometry, and polarimetry observations of Saturn, its rings, and Titan. A new ring of optical depth < 2 x 10(-3) was discovered at 2.33 Saturn radii and is provisionally named the F ring; it is separated from the A ring by the provisionally named Pioneer division. A division between the B and C rings, a gap near the center of the Cassini division, and detail in the A, B, and C rings have been seen; the nomenclature of divisions and gaps is redefined. The width of the Encke gap is 876 +/- 35 kilometers. The intensity profile and colors are given for the light transmitted by the rings. A mean particle size less, similar 15 meters is indicated; this estimate is model-dependent. The D ring was not seen in any viewing geometry and its existence is doubtful. A satellite, 1979 S 1, was found at 2.53 +/- 0.01 Saturn radii; the same object was observed approximately 16 hours later by other experiments on Pioneer 11. The equatorial radius of Saturn is 60,000 +/- 500 kilometers, and the ratio of the polar to the equatorial radius is 0.912 +/- 0.006. A sample of polarimetric data is compared with models of the vertical structure of Saturn's atmosphere. The variation of the polarization from the center of the disk to the limb in blue light at 88 degrees phase indicates that the density of cloud particles decreases as a function of altitude with a scale height about one-fourth that of the gas. The pressure level at which an optical depth of 1 is reached in the clouds depends on the single-scattering polarizing properties of the clouds; a value similar to that found for the Jovian clouds yields an optical depth of 1 at about 750 millibars.

First Report of Sclerotium rolfsii on Ascocentrum and Ascocenda Orchids in Florida.

Southern blight caused by Sclerotium rolfsii is known to occur on several economically important orchid hosts, including Vanda species and hybrids (1-3). In the summer and fall of 2008, an outbreak of southern blight on Vanda orchids was seen in several commercial nurseries and landscapes throughout South Florida. More than a dozen orchids were affected at one of the locations, and symptoms of S. rolfsii were observed on Ascocentrum and Ascocenda orchids, which are also common in the trade and demand a resale value ranging from $20 to $150 for specimens in bloom. Affected Ascocentrum and Ascocenda orchids were found severely wilted at the apex, while around the base of the plants, tan, soft, water-soaked lesions were present. As the lesions progressed, leaves around the base of the plants began to fall off, leaving the stems bare. After 2 days, white, flabellate mycelium was seen progressing up the stem and numerous, tan-to-brown sclerotia were present. Leaves and portions of the stems were plated on acidified potato dextrose agar (APDA) and grown at 25°C. White, flabellate mycelium and tan sclerotia approximately 2 mm in diameter were produced in culture and microscopic examination revealed the presence of clamp connections. The fungus was identified as S. rolfsii and a voucher specimen was deposited with the ATCC. A PCR was performed on the ITS1, 5.8S rDNA, and ITS2 and the sequence was deposited in GenBank (Accession No. GQ358518). Pathogenicity of an isolate was tested by placing 6-mm plugs taken from APDA plates directly against the stem of five different Ascocentrum and Ascocenda orchids. Five Ascocentrum and Ascocenda orchids were inoculated with 6-mm plugs of plain APDA and five were untreated controls. Plants were housed under 50% shade, 60 to 95% humidity, and temperatures ranging from 75 to 88°F. Within 7 days, all inoculated plants developed symptoms that were identical to those observed on original plants and S. rolfsii was consistently reisolated from symptomatic tissue. Ascocentrum and Ascocenda were previously reported under miscellaneous orchid species and hybrids as hosts for S. rolfsii (1). However, this report was highly ambiguous and the most current edition does not report the host fungus combination (2). To our knowledge, this is the first report of S. rolfsii affecting Ascocentrum and Ascocenda orchids. References: (1) S. A. Alfieri, Jr., et al. Diseases and Disorders of Plants in Florida. Bull. No. 11. Division of Plant Industry, Gainesville, FL, 1984. (2) S. A. Alfieri, Jr., et al. Diseases and Disorders of Plants in Florida. Bull. No. 14. Division of Plant Industry, Gainesville, FL, 1994. (3) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989.

First Report of a Bacterial Soft Rot on Tolumnia Orchids Caused by a Dickeya sp. in the United States.

Tolumnia orchids are small epiphytic orchids grown for their attractive flowers. In the fall of 2008, approximately 100 Tolumnia orchids with soft, brown, macerated leaves were brought to the University of Florida Extension Plant Diagnostic Clinic in Homestead. Ten plants were randomly selected and bacteria were isolated from the margins of symptomatic tissues of each of the 10 plants on nutrient agar according to the method described by Schaad et al. (2). Four reference strains were used in all tests, including the molecular tests: Erwinia carotovora subsp. carotovora (obtained from J. Bartz, Department of Plant Pathology, University of Florida, Gainesville), E. chrysanthemi (ATCC No. 11662), Pectobacterium cypripedii (ATCC No. 29267), and Acidovorax avenae subsp. cattleyae (ATCC No. 10200). All 10 of the isolated bacteria were gram negative, grew at 37°C, degraded pectate in CVP (crystal violet pectate) medium, grew anaerobically, produced brown pigment on NGM (nutrient agar-glycerol-manganese chloride) medium (1), were sensitive to erythromycin, and produced phosphatase. Three of the strains were submitted for MIDI analysis (Sherlock version TSBA 4.10; Microbial Identification, Newark DE) (SIM 0.732 to 0.963), which identified them as E. chrysanthemi. A PCR assay was performed on the 16S rRNA gene with primers 27f and 1495r described by Weisburg et al. (3) from two of the isolates and a subsequent GenBank search showed 99% identity of the 1,508-bp sequence to that of Dickeya chrysanthemi (Accession No. FM946179) (formerly E. chrysanthemi). The sequences were deposited in GenBank (Accession Nos. GQ293897 and GQ293898). Pathogenicity was confirmed by injecting approximately 100 µl of a bacterial suspension at 1 × 108 CFU/ml into leaves of 10 Tolumnia orchid mericlones. Ten plants were also inoculated with water as controls. Plants were placed in a greenhouse at 29°C with 60 to 80% relative humidity. Within 24 h, soft rot symptoms appeared on all inoculated leaves. The water controls appeared normal. A Dickeya sp. was reisolated and identified using the above methods (biochemical tests and MIDI), fulfilling Koch's postulates. To our knowledge, this is the first report of a soft rot caused by a Dickeya sp. on Tolumnia orchids. Although 16S similarity and MIDI results suggest the isolated bacteria are D. chrysanthemi because of its close similarity with other Dickeya spp., these results are not conclusive. Further work should be conducted to confirm the identity of these isolates. Through correspondence with South Florida Tolumnia growers, it appears this disease has been a recurring problem, sometimes affecting international orchid shipments where plant losses have been in excess of 70%. References: (1) Y. A. Lee and C. P. Yu. J. Microbiol. Methods 64:200, 2006. (2) N. W. Schaad et al. Erwinia soft rot group. Page 56 in: Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed. N. W. Schaad et al., eds. American Phytopathological Society. St. Paul, MN, 2001. (3) W. G. Weisburg et al. J. Bacteriol. 173:697, 1991.

Listening to women with intellectual disabilities and mental health problems: a focus on risk and resilient factors.

Women without intellectual disabilities are more likely to develop mental health problems as a result of physiological functioning and psychosocial risk factors. However, little is known about the mental health of women with intellectual disabilities. The aim of this study was to explore a small group of women's perceptions of the risk and protective factors pertaining to their mental health conditions. Twelve semi-structured interviews were conducted in 2007 in Northern Ireland. Thematic content analysis identified three risk factors and four protective/resilient factors. None of the women identified physiological functioning as a risk factor. Results suggest that women with intellectual disabilities experience psychosocial risk factors similar to those reported by women without intellectual disability. Additional risk factors place them at higher risk of developing mental health problems. However, more research is required.

Day-case holmium laser enucleation of the prostate: feasibility, safety and predictive factors.

Introduction Transurethral resection of the prostate has remained the most common operation for bladder outlet obstruction in the UK, but it is associated with potential morbidity and median two-day length of hospital stay. Holmium laser enucleation of the prostate (HoLEP) provides an alternative procedure. Provision of day-case HoLEP would improve patient care through increased efficiency. We assessed the feasibility and safety of day-case HoLEP and examined predictive factors for increased length of hospital stay. Materials and methods Patients presenting for HoLEP by a single surgeon from September 2013 to September 2016 were considered for day-case surgery. Patients were discharged following assessment by the operating surgeon and met predetermined discharge criteria. Factors contributing to day-case success were identified. Results In total, 210 patients (mean age 70.3 ± 8.5 years) underwent HoLEP, with 74 (35.3%) discharged as true day-cases and a further 84 (40.0%) discharged within 23 hours. Readmission rate was 5.5%, with all complications Clavien-Dindo grade I or II. Factors associated with successful day-case operation included low-volume prostates (≤ 40 g) (odds ratio, OR, 3.097, 95% confidence interval, CI, 1.619-5.924, P = 0.0001) and morning surgical lists (OR 6.124, 95% CI 2.526-14.845, p<0.001). Discussion and conclusion Day-case HoLEP is both feasible and safe, with low readmission rates. Two factors were significantly associated with successful day-case surgery: small volume prostate and morning theatre lists. Addressing these factors through preoperative planning can improve day-case surgery rates and improve bed throughput.

Molecular cloning and preliminary characterization of a novel cytoplasmic antigen recognized by myasthenia gravis sera.

A cDNA clone was isolated by screening of a lambda gt11 endothelial expression library with serum from a patient with myasthenia gravis (MG). Rabbit antisera raised against the recombinant protein and human MG sera reactive with the clone immunoblotted an M(r) integral of 250,000 polypeptide (gravin) present in endothelial cells and several adherent cells. Gravin was not detected in platelets, leukocytes, U937, or human erythroleukemic (HEL) cell lines, but was expressed in HEL cells after induction with phorbol myristate acetate. Northern blot analysis showed two transcripts of approximately 6.7 and 8.4 kb in endothelial cells but not U937 or HEL cells. Indirect immunofluorescence of permeabilized cells revealed a trabecular network of gravin staining with a distinct linear component. Antibodies to gravin, were present in sera from 22:72 (31%) of MG patients. In contrast 0:50 normal sera and 1:72 sera from patients with other autoimmune diseases contained antigravin antibodies. Gravin is not likely to be a nonerythroid spectrin, talin, myosin, or actin-binding protein based on the lack of reactivity of antigravin with these polypeptides in immunoblots. The nucleotide sequence of the immunoreactive clone indicated that it encodes a highly acidic polypeptide fragment that contains the carboxyl terminus of the protein. Neither amino acid nor nucleotide sequences were present in Genbank, EMBL, or Swissprot databases as of March, 1992. These data indicate that gravin is an inducible, cell type-specific cytoplasmic protein and that auto-antibodies to gravin may be highly specific for MG.

Rheumatoid factor-producing cells detected by direct hemolytic plaque assay.

Lymphocytes secreting anti-IgC antibodies, rheumatoid factors (RF), can be detected in the peripheral bloods, synovial fluids, and bone marrows of patients with seropositive rheumatoid arthritis by using a direct plaque-forming cell (PFC) assay with sheep erythrocytes sensitized with reduced and alkylated rabbit IgG hemolysin. The autospecific nature of the RF produced by RF-PFC was indicated by inhibition studies in which the order of patency was human IgG greater than rabbit IgG greater than bovine IgG. In metabolic studies puromycin, cycloheximide, and venblastine suppressed RF-PFC. Cyclic AMP and cyclic GMP were without effect. A need was recognized for using full tissue culture media during the cell separation and plaquing procedures to optimize detection of the RF-PFC. RF-PFC may appear in the blood of patients intermittently despite their continuing presence in the bone marrow. They have been found in the peripheral blood, especially during acutely exacerbating polyarticular synovitis, generalized vasculities, or generally active, aggressive disease. RF-PFC were found in synovial effusions of new or recrduescent acute synovitis. RF-PFC were observed to disappear from the peripheral circulation and the bone marrow during therapy with cytotoxic drugs. The data are consistent with the hypothesis that the appearance of RF-PFC in the peripheral blood represents an anamnestic response to transiently appearing antigen. The nature of the antigen is not specified. The bone marrow may be a site of origin of RF-PFC.

Nitrogen mineralization and microbial activity in oil sands reclaimed boreal forest soils.

Organic materials including a peat-mineral mix (PM), a forest floor-mineral mix (L/S), and a combination of the two (L/PM) were used to cap mineral soil materials at surface mine reclamation sites in the Athabasca oil sands region of northeastern Alberta, Canada. The objective of this study was to test whether LFH provided an advantage over peat by stimulating microbial activity and providing more available nitrogen for plant growth. Net nitrification, ammonification, and N mineralization rates were estimated from field incubations using buried bags. In situ gross nitrification and ammonification rates were determined using the 15N isotope pool dilution technique, and microbial biomass C (MBC) and N (MBN) were measured by the chloroform fumigation-extraction method. All reclaimed sites had lower MBC and MBN, and lower net ammonification and net mineralization rates than a natural forest site (NLFH) used as a control, but the reclamation treatment using LFH material by itself had higher gross and net nitrification rates. A positive correlation between in situ moisture content, dissolved organic N, MBC, and MBN was observed, which led us to conduct a moisture manipulation experiment in the laboratory. With the exception of the MBN for the L/S treatment, none of the reclamation treatments ever reached the levels of the natural site during this experiment. However, materials from reclamation treatments that incorporated LFH showed higher respiration rates, MBC, and MBN than the PM treatment, indicating that the addition of LFH as an organic amendment may stimulate microbial activity as compared to the use of peat alone.

Orofacial pain - an update on diagnosis and management.

The diagnosis and management of orofacial pain may be challenging due to complex histories, pathophysiology and associated psychosocial co-morbidities such as depression and anxiety. Neuropathic facial pain conditions such as burning mouth syndrome (BMS), persistent idiopathic facial pain (PIFP), atypical odontalgia (AO) and trigeminal neuralgia (TN) require early recognition by primary care clinicians and referral to secondary care. Acute pain-related temporomandibular disorder (TMD) may be managed in the primary care setting, with identification of those at risk of developing chronic TMD receiving an early referral to secondary care. Adopting a biopsychosocial approach, consisting of physical therapies, pharmacotherapy and psychological support can lead to effective management and may limit the negative impact of facial pain upon quality of life and daily functioning.

Metabolism of unsaturated fatty acids by RBL-1 5-lipoxygenase: influence of substrate solubility and product inactivation.

Several alternative fatty acid substrates have been employed to characterise the kinetics of rat basophilic leukaemia cell (RBL-1) 5-lipoxygenase. Using arachidonic acid (AA) as substrate, enzymes rates declined at high substrate concentrations (greater than 25 microM) and were associated with pronounced lag phases. The concentrations of AA at which apparent substrate inhibition and lag phases were observed were comparable with those at which AA induced emulsion formation in aqueous media. No evidence for substrate inhibition or lag phases was observed using eicosapentaenoic acid (EPA), a more soluble substrate which did not induce emulsion formation at concentrations up to 100 microM. Reactions catalysed by RBL-1 5-lipoxygenase terminated before exhaustion of substrate. AA and EPA induced time-dependent enzyme inactivation at concentrations 100-fold lower than their apparent Km values for the enzyme. The ability of several fatty acids to induce time-dependent inactivation was directly proportional to their substrate potency. We conclude that apparent substrate inhibition is a consequence of a change from monomeric to micellar substrate which has a lower affinity for the enzyme and that premature termination of the enzyme reactions is a consequence of product-induced enzyme inactivation.