Opioid Analgesics Do Not Improve Abdominal Pain or Quality of Life in Crohn's Disease.

BACKGROUND: Abdominal pain and opioid analgesic use are common in Crohn's disease (CD). AIMS: We sought to identify factors associated with abdominal pain in CD and evaluate the impact of opioid analgesics on pain and quality-of-life scores in this setting. METHODS: We performed a longitudinal cohort study using a prospective, consented IBD natural history registry from a single academic center between 2009 and 2013. Consecutive CD patients were followed for at least 1 year after an index visit. Data were abstracted regarding pain experience (from validated surveys), inflammatory activity (using endoscopic/histologic findings), laboratory studies, coexistent psychiatric disorders, medical therapy, opioid analgesic, and tobacco use. RESULTS: Of 542 CD patients (56.6% women), 232 (42.8%) described abdominal pain. Individuals with pain were more likely to undergo surgery and were more frequently prescribed analgesics and/or antidepressants/anxiolytics. Elevated ESR (OR 1.79; 95%CI 1.11-2.87), coexistent anxiety/depression (OR 1.87; 95%CI 1.13-3.09), smoking (OR 2.08; 95%CI 1.27-3.40), and opioid use (OR 2.46; 95%CI 1.33-4.57) were independently associated with abdominal pain. Eighty patients (14.8%) were prescribed opioids, while 31 began taking them at or after the index visit. Patients started on opioids demonstrated no improvement in abdominal pain or quality-of-life scores on follow-up compared to patients not taking opioids. CONCLUSIONS: Abdominal pain is common in CD and is associated with significant opioid analgesic utilization and increased incidence of anxiety/depression, smoking, and elevated inflammatory markers. Importantly, opioid use in CD was not associated with improvement in pain or quality-of-life scores. These findings reinforce the limitations of currently available analgesics in IBD and support exploration of alternative therapies.

The modular structure of actin-regulatory proteins.

Filamentous actin structures possess unique biophysical and biochemical properties and are required for cell locomotion, cell division, compartmentalization and morphological processes. The site-specific assembly and disassembly of these structures are directed by actin-regulatory proteins. This article reviews how structural studies are now defining the atomic details of small modular domains present in actin-regulatory proteins responsible for crosslinking, severing and capping of actin filaments, as well as for localization of actin filament assembly. These studies have identified three modular strategies for the design of proteins that regulate the actin cytoskeleton.

Isolation and identification of ochratoxin A-producing Aspergillus section Nigri strains from California raisins.

AIMS: To determine incidence and levels of ochratoxin A (OTA) in California raisins and to isolate and characterize OTA-producing fungi from California raisin vineyard populations. METHODS AND RESULTS: Forty raisin clusters sampled from four California vineyards in the San Joaquin Valley were analysed for OTA content using immunoaffinity and HPLC methods. OTA was detected in 93% of the samples, at levels from 0·06 to 11·4 ng g⁻¹. From these raisin samples, a total of 400 strains of Aspergillus were isolated and analysed for OTA production. Twelve isolates (3%), from five raisin samples, produced OTA. These isolates were identified as Aspergillus carbonarius, based on morphological characteristics and multilocus sequence analysis. Levels of OTA produced by these isolates on raisin agar ranged from 0·9 to 15 µg g⁻¹. CONCLUSIONS: OTA is a common contaminant of raisin vineyards, but average levels are much lower than EU regulatory limits for dried fruit. The primary species responsible for OTA contamination in California raisins is A. carbonarius. SIGNIFICANCE AND IMPACT OF THE STUDY: This study illustrates that low-level OTA contamination of raisins occurs in California and that ecological studies of A. carbonarius within the Aspergillus section Nigri population on raisins are warranted to monitor ochratoxigenic potential of the crop.

Use of chemosensitization to overcome fludioxonil resistance in Penicillium expansum.

AIM: To overcome fludioxonil resistance of Penicillium expansum, a mycotoxigenic fungal pathogen causing postharvest decay in apple, by using natural phenolic chemosensitizing agents. METHODS AND RESULTS: Fludioxonil-resistant mutants of P. expansum were co-treated with different oxidising and natural phenolic agents. Resistance was overcome by natural phenolic chemosensitizing agents targeting the oxidative stress-response pathway. These agents also augmented effectiveness of the fungicide, kresoxim-methyl. Results indicated that alkyl gallates target mitochondrial respiration and/or its antioxidation system. Fungal mitochondrial superoxide dismutase (Mn-SOD) plays a protective role against alkyl gallates. CONCLUSIONS: Natural chemosensitizing agents targeting the oxidative stress-response system, such as Mn-SOD, can synergize commercial fungicides. SIGNIFICANCE AND IMPACT OF THE STUDY: Redox-active compounds can serve as potent chemosensitizing agents to overcome resistance and lower effective dosages of fungicides. This can reduce costs with coincidental lowering of environmental and health risks.

In vivo importance of actin nucleotide exchange catalyzed by profilin.

The actin monomer-binding protein, profilin, influences the dynamics of actin filaments in vitro by suppressing nucleation, enhancing nucleotide exchange on actin, and promoting barbed-end assembly. Profilin may also link signaling pathways to actin cytoskeleton organization by binding to the phosphoinositide PIP(2) and to polyproline stretches on several proteins. Although activities of profilin have been studied extensively in vitro, the significance of each of these activities in vivo needs to be tested. To study profilin function, we extensively mutagenized the Saccharomyces cerevisiae profilin gene (PFY1) and examined the consequences of specific point mutations on growth and actin organization. The actin-binding region of profilin was shown to be critical in vivo. act1-157, an actin mutant with an increased intrinsic rate of nucleotide exchange, suppressed defects in actin organization, cell growth, and fluid-phase endocytosis of pfy1-4, a profilin mutant defective in actin binding. In reactions containing actin, profilin, and cofilin, profilin was required for fast rates of actin filament turnover. However, Act1-157p circumvented the requirement for profilin. Based on the results of these studies, we conclude that in living cells profilin promotes rapid actin dynamics by regenerating ATP actin from ADP actin-cofilin generated during filament disassembly.

Pathogenesis of Eutypa lata in grapevine: identification of virulence factors and biochemical characterization of cordon dieback.

Eutypa lata is a vascular pathogen of woody plants. In the present study we (i) determined which component(s) of the cell wall polymers were degraded in naturally infected grapevines and in artificially inoculated grape wood blocks; (ii) compared the pattern of wood decay in the tolerant grape cv. Merlot versus the susceptible cv. Cabernet Sauvignon; and (iii) identified secondary metabolites and hydrolytic enzymes expressed by E. lata during wood degradation. Biochemical analyses and a cytochemical study indicated that glucose-rich polymers were primary targets of E. lata. Structural glucose and xylose of the hemicellulose fraction of the plant cell wall and starch were depleted in infected woods identically in both cultivars. Moreover, the more tolerant cv. Merlot always had more lignin in the wood than the susceptible cv. Cabernet Sauvignon, indicating that this polymer may play a role in disease resistance. In vitro assays demonstrated the production by E. lata of oxidases, glycosidases and starch degrading enzymes. Phytotoxic secondary metabolites were also produced but our data suggest that they may bind to the wood. Finally, we demonstrated that free glucose in liquid cultures repressed primary but not secondary metabolism.

Evaluation of Therapeutics for Advanced-Stage Heart Failure and Other Severely-Debilitating or Life-Threatening Diseases.

Severely-debilitating or life-threatening (SDLT) diseases include conditions in which life expectancy is short or quality of life is greatly diminished despite available therapies. As such, the medical context for SDLT diseases is comparable to advanced cancer and the benefit vs. risk assessment and development of SDLT disease therapeutics should be similar to that of advanced cancer therapeutics. A streamlined development approach would allow patients with SDLT conditions earlier access to therapeutics and increase the speed of progression through development. In addition, this will likely increase the SDLT disease therapeutic pipeline, directly benefiting patients and reducing the economic and societal burden of SDLT conditions. Using advanced-stage heart failure (HF) as an example that illustrates the concepts applicable to other SDLT indications, this article proposes a streamlined development paradigm for SDLT disease therapeutics and recommends development of aligned global regulatory guidance.

A Reassessment of the Species Concept in Eutypa lata, the Causal Agent of Eutypa Dieback of Grapevine.

ABSTRACT Eutypa dieback is a vascular disease of several cultivated crops and trees worldwide. The attribution of the name to the agent responsible for branch dieback is ambiguous. Pathogenicity of Eutypa sp. first was reported on apricot and the causal agent was named E. armeniacae. However, no morphological differences were reported with the previously described E. lata, and some authors considered both species synonymous. Others regarded them as distinct species on the basis of pathogenesis and molecular analysis. We further investigated the relatedness of both species by phylogenetic analyses of the internal transcribed spacer region and beta-tubulin gene. These analyses included several other taxa placed in the same family (Diatrypaceae), and yielded three groups. The isolates referred to as E. lata in previous work clustered with Diatrype stigma in one group. Isolates of E. armeniacae and E. lata clustered in a second group, supporting the synonymy of these species. The third group included other Eutypa spp. supporting the polyphyletic origin of this genus. Measurements of conidia length and secondary metabolite production of isolates supported the phylogenetic analyses. Secondary metabolites appeared to be a synapomorphic character shared by several taxa including E. lata, E. armeniacae, E. laevata, and E. petrakii var. petrakii.

The molecular basis for allergen cross-reactivity: crystal structure and IgE-epitope mapping of birch pollen profilin.

BACKGROUND: The profilins are a group of ubiquitous actin monomer binding proteins that are responsible for regulating the normal distribution of filamentous actin networks in eukaryotic cells. Profilins also bind polyphosphoinositides, which can disrupt the profilin-action complex, and proline-rich ligands which localize profilin to sites requiring extensive actin filament accumulation. Profilins represent cross-reactive allergens for almost 20 % of all pollen allergic patients. RESULTS: We report the X-ray crystal structure of birch pollen profilin (BPP) at 2.4 resolution. The major IgE-reactive epitopes have been mapped and were found to cluster on the N- and C-terminal alpha helices and a segment of the protein containing two strands of the beta sheet. The overall fold of this protein is similar to that of the mammalian and amoeba profilins, however, there is a significant change in the orientation of the N-terminal alpha helix in BPP. This change in orientation alters the topography of a hydrophobic patch on the surface of the molecule, which is thought to be involved in the binding of proline-rich ligands. CONCLUSIONS: Profilin has been identified as an important cross-reactive allergen for patients suffering from multivalent type I allergy. The prevalent epitopic areas are located in regions with conserved sequence and secondary structure and overlap the binding sites for natural profilin ligands, indicating that the native ligand-free profilin acts as the original cross-sensitizing agent. Structural homology indicates that the basic features of the G actin-profilin interaction are conserved in all eukaryotic organisms, but suggests that mechanistic differences in the binding of proline-rich ligands may exist. The structure of BPP provides a molecular basis for understanding allergen cross-reactivity.

Blocking the ends of transforming DNA enhances gene targeting in Dictyostelium.

Eukaryotic gene targeting by means of gene replacement vectors is often complicated by unwanted plasmid insertion events involving the ends of transforming DNA molecules. These undesirable and often multiple insertions occur both randomly (i.e. non-homologously) and at the targeted locus. By blocking the 3' ends of transforming DNA with 2'3' dideoxynucleotides, we have reduced the frequency of end-mediated DNA insertion in Dictyostelium amoebae. As a result, only one copy of the selectable gene is introduced at the target locus to achieve a precise gene disruption.

Cognitive and physical capacity process variables predict long-term outcome after treatment of chronic pain.

Cognitive-behavioral and physical therapies are incorporated into multidisciplinary chronic pain programs because changes in pain cognitions and physical capacity may represent therapeutic processes that facilitate favorable outcome. Decreases in depression, however, may explain treatment responses more parsimoniously. Measures of pain helplessness, lifting capacity, walking endurance, depression, pain severity, and activity level were collected from 94 chronic pain patients at pre- and posttreatment and at 3- to 6-month follow-up evaluations. Decreases in pain helplessness were linked to pain severity reduction, whereas walking endurance increases were related to improvements in activity levels and downtime even after controlling for effects of depression decreases. Thus, cognitive and physical capacity changes that occur through pain treatment may make unique contributions to long-term outcome.

Association between workers' compensation and outcome following multidisciplinary treatment for chronic pain: roles of mediators and moderators.

OBJECTIVE: To determine whether the tendency for chronic pain patients who receive Workers' Compensation to show a poorer response than non-compensated patients to pain treatment can be accounted for by mediating factors; to assess whether moderating factors can distinguish subgroups of Workers' Compensation recipients who react very poorly to treatment from compensated patients who respond well. DESIGN: Outcome study based on archives. SETTING: Multidisciplinary pain treatment center. PATIENTS: Of 214 patients, 158 had complete data. OUTCOME MEASURE: Blind ratings of narrative discharge summaries written by the Pain Treatment Center staff. RESULTS: A significant negative relationship between receiving Workers' Compensation and outcome was mediated by a pessimistic belief in the ability to return to former occupation. Moderator effects showed that Workers' Compensation recipients with high initial pain and a history of pain-related surgery fared worse than any other group. Moreover, Workers' Compensation recipients not characterized by high pain and a history of surgery responded as well as noncompensated patients. CONCLUSIONS: The inadequate response to pain programs shown by Workers' Compensation recipients may be partly understood in terms of well-defined mediating factors, which may admit to amelioration via clinical intervention. Moreover, Workers' Compensation patients should not be considered high risks for failure by sole virtue of their compensation status. Multifactor assessment methods may be needed to identify that portion of compensation recipients who are actually at appreciable risk for treatment failure so that appropriate adjustments in treatment regimen may be made.

Oxidation of acetaldehyde and presence of aldehyde dehydrogenase in rat erythrocytes.

Rat liver erythrocytes were found to oxidize acetaldehyde at 7 nmoles/min/ml blood at 37 degrees C. This is less than 1% the rate that occurs in liver. An aldehyde dehydrogenase was isolated from erythrocytes, but was not purified. The enzyme had a Km of 170 microM toward acetaldehyde at pH 7.4. The enzyme, which could oxidize both aliphatic and aromatic aldehydes, was more active at pH 9 than at 7. Disulfiram proved to be both an in vivo and in vitro inhibitor of the enzyme. Due to the low total capacity of the erythrocytes to metabolize acetaldehyde, it is doubtful they perform any important role in ethanol metabolism.

Anger management style and the prediction of treatment outcome among male and female chronic pain patients.

Anger is a prominent emotion experienced by chronic pain patients. Anecdotes suggest that anger predicts poor outcome following multidisciplinary pain programs, but no empirical evidence documents this link. We expected that patient anger expression or suppression would predict poor outcome following a pain program and that gender differences would emerge. Pre- to posttreatment measures of lifting capacity, walking endurance, depression, pain severity and activity level were collected from 101 chronic pain patients. An 'anger expression x gender' interaction was found such that anger expression among males was correlated negatively with lifting capacity improvements. 'Anger suppression x gender' interactions emerged such that anger suppression among males was correlated negatively with improvements in depression and general activities. These effects remained significant after controlling for trait anger. Thus, how anger is managed may exert unique influence on outcomes apart from the effects of mere anger proneness, at least among male pain patients.

Regulation of aflatoxin production by naphthoquinones of walnut (Juglans regia).

Walnuts are a valuable crop the sale and export potential of which may be severely limited by contamination with aflatoxins, metabolites produced on infection with Aspergillus flavus. The effect of a series of four naphthoquinones [1,4-naphthoquinone (1); juglone (5-hydroxy-1,4-naphthoquinone) (2); 2-methyl-1, 4-naphthoquinone (3); and, plumbagin (5-hydroxy-2-methyl-1, 4-naphthoquinone) (4)] (Figure 1), which occur in walnut husks, on fungal viability and aflatoxigenesis was studied in vitro. The quinones delayed germination of the fungus and were capable of completely inhibiting growth at higher concentrations. Their effect on aflatoxin levels was highly dependent on the concentration of individual naphthoquinones in the media. At higher concentrations, aflatoxin production was decreased or completely inhibited, but at lower concentrations there was a stimulatory effect on aflatoxin biosynthesis, with a >3-fold increase at 20 ppm of 3. Structural features associated with decreased fungal viability and greatest effect on aflatoxigenesis are the presence of a 5-hydroxyl or 2-methyl substituent, but there is no significant additive effect when both of these substituents are present.

Identification of benzethonium chloride in commercial grapefruit seed extracts.

Commercial grapefruit seed extracts (GSE) were extracted with chloroform. The solvent was evaporated, and the resulting solid was subsequently analyzed by high-performance liquid chromatography, electrospray ionization mass spectrometry, nuclear magnetic resonance (NMR) spectroscopy, and elemental analysis (by proton-induced X-ray emission [PIXE] analysis). The main constituent was identified as benzethonium chloride, a synthetic antimicrobial agent commonly used in cosmetics and other topical applications. This compound comprised 8.03% (n = 2) of the liquid GSE sample. Higher amounts of benzethonium chloride were found in powder GSE samples.

Coordinator teaches staff to operate equipment, handles malfunctioning equipment.

Based on the data collection, a recommendation was made to the OR committee to develop a new full-time position of equipment coordinator. Committee members supported the recommendation, and further recommended that a backup person be trained as an assistant equipment coordinator to cover when the equipment coordinator was not available.

Benign recurrent sixth (abducens) nerve palsies in children.

Sixth nerve palsy can occur as a result of elevated intracranial pressure, neoplasm or trauma. Reports from tertiary centres indicate that between 5% and 16% of referred cases have no ascribed aetiology and are classified as benign. Rarely, these benign palsies can recur. A retrospective chart review of a cohort of 253 paediatric patients with sixth nerve palsies was analysed and uncovered 30 cases of benign sixth nerve palsy, nine of which recurred. Our data and review of other studies on the subject imply that a new onset sixth nerve palsy presenting in children can be benign in approximately 13% of cases, so a thorough history and physical examination to evaluate for any other neurological symptoms or signs followed by MRI of the brain with and without contrast is recommended.

Enhanced activity of strobilurin and fludioxonil by using berberine and phenolic compounds to target fungal antioxidative stress response.

AIMS: Identify natural products that effectively target antioxidative signal transduction/stress response systems [i.e., mitogen-activated protein kinase (MAPK) pathway, mitochondrial superoxide dismutase (Mn-SOD)] of fungi. Enhance activity of strobilurin or fludioxonil with discovered compounds. METHODS AND RESULTS: Enhancement of antifungal activity of strobilurins, inhibitors of complex III of the mitochondrial respiratory chain, was tested using berberine hemisulfate and different phenolic compounds. The Saccharomyces cerevisiae sod2Delta, a deletion mutant lacking Mn-SOD gene, was highly sensitive to berberine and veratraldehyde. Functional complementation analysis verified these compounds target Mn-SOD. Activity of strobilurin (25-50 micromol l(-1)) was elevated on most aspergilli and Penicillium expansum by co-application with berberine or veratraldehyde (2-4 mmol l(-1)). These compounds also prevented Aspergillus fumigatus MAPK mutants (sakADelta and mpkCDelta) from escaping toxicity of fludioxonil (a phenylpyrrole fungicide potentiated by the MAPK pathway), a typical phenotype of fungal MAPK mutants. CONCLUSIONS: Strobilurin activity or prevention of fungal escape from fludioxonil toxicity can be enhanced by co-application of certain alkaloids or phenolics. SIGNIFICANCE AND IMPACT OF THE STUDY: Natural products can be used to target cellular stress response systems in fungal pathogens and serve as alternatives/additives to commercial antifungal agents.

Targeting antioxidative signal transduction and stress response system: control of pathogenic Aspergillus with phenolics that inhibit mitochondrial function.

AIMS: The aim of this study was to show whether antioxidative response systems are potentially useful molecular targets for control of Aspergillus fumigatus and Aspergillus flavus. Selected phenolic agents are used in target-gene-based bioassays to determine their impact on mitochondrial respiration. METHODS AND RESULTS: Vanillyl acetone, vanillic acid, vanillin, cinnamic acid, veratraldehyde, m-coumaric acid (phenolic agents to which Saccharomyces cerevisiae sod2delta mutant showed sensitivity), carboxin (inhibits complex II of the mitochondrial respiratory chain), strobilurins/antimycin A (inhibits complex III of the mitochondrial respiratory chain) and fludioxonil/fenpiclonil [antifungals potentiated by mitogen-activated protein kinase (MAPK)] were examined in A. fumigatus, A. flavus and S. cerevisiae. Individual or combined application of phenolics with inhibitors of mitochondrial respiration showed some of the phenolics effectively inhibited fungal growth. Target-gene bioassays were performed using a sakAdelta (MAPK deletion) strain of A. fumigatus and a complementation analysis using the mitochondrial superoxide dismutase (Mn-SOD) gene (sodA) of A. flavus in the ortholog mutant, sod2delta, of S. cerevisiae. The results demonstrated that mitochondrial antioxidative stress system plays important roles in fungal response to antifungal agents tested. CONCLUSIONS: Antioxidative response systems of fungi can be an efficient molecular target of phenolics for pathogen control. Combined application of phenolics with inhibitors of mitochondrial respiration can effectively suppress the growth of fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: Natural compounds that do not pose any significant medical or environmental risks could serve as useful alternatives or additives to conventional antifungals. Identifying the antioxidative response systems in other pathogens could improve methods for fungal control.