Research Note: Evaluation of phytic acid disappearance, ileal P digestibility, and total tract P retention in canola meal supplemented with increasing levels of exogenous phytase using conventional and cecectomized precision-fed roosters and growing chicks.

Apparent ileal digestibility (AID) of P, apparent total tract retention (ATTR) of P, and phytic acid disappearance in canola meal were evaluated in the presence of increasing levels of exogenous phytase. In Experiment 1, a precision-fed rooster assay was used to determine phytic acid (myo-inositol 1,2,3,4,5,6-hexakis; InsP6) and inositol phosphate (InsP6-3; InsP-P) disappearance in conventional and cecectomized Leghorn roosters. Roosters were crop intubated with 25 g of canola meal mixed with 0, 500, 1,000, or 2,000 FTU/kg of exogenous phytase. In Experiment 2, InsP6 and InsP-P disappearance and AID and ATTR of P were determined using ad libitum-fed broiler chickens. Treatments consisted of semi-purified diets containing 45% canola meal as the sole source of P. Phytase was added to increase phytase activity by 0, 500, 1,000, or 2,000 FTU/kg. Experiments contained 6 replicates per treatment. Canola meal contained a high phytase activity (1,630 FTU/kg as-fed) due to contamination with a commercially available phytase at the feed mill from which the canola meal was sourced. In Experiment 1 with precision-fed roosters, there was no effect (P > 0.05) of phytase or bird type on InsP6 and InsP-P disappearance; however, phytase linearly reduced (P < 0.05) InsP3 concentrations in excreta. In Experiment 2 with ad libitum-fed chickens, phytase linearly increased (P < 0.05) ileal InsP6 and InsP-P disappearance, and phytase had a quadratic effect (P < 0.05) on excreta InsP6 and InsP-P disappearance. Increasing dietary phytase activity resulted in a linear increase (P < 0.05) in AID of P and phytase had a quadratic effect (P < 0.05) on ATTR of P. In conclusion, titration of high levels of phytase (1,600 to 3,600 FTU/kg as-fed) reduced InsP3 concentrations in precision-fed roosters but did not affect overall phytic acid hydrolysis, which was 78% or greater for all treatments; however, increasing the total phytase activity from 700 to 2,700 FTU in ad libitum-fed broiler chickens increased phytic acid disappearance and P digestibility.

Research Note: Relative bioavailability of zinc in zinc hydroxychloride for chicks.

A chick assay was conducted to determine the effects of Zn source on performance and to establish a Zn relative bioavailability value (RBV) for a new source of Zn hydroxychloride. In the assay, 8-day-old chicks were fed a Zn-deficient soy protein concentrate diet supplemented with 0, 7, and 15 mg Zn/kg from feed grade ZnSO4 monohydrate for 14 d to establish a standard response curve. The same basal diet was supplemented with 3, 7, and 10 mg Zn/kg from a new Zn hydroxychloride (SAMZn). A second source of Zn hydroxychloride (IBZn) was supplemented at 10 mg Zn/kg as a direct comparison to the highest level of SAMZn. Weight gain increased (P < 0.05) with increasing Zn level, regardless of source. Weight gain of chicks fed 7 mg Zn/kg from SAMZn was not different (P > 0.05) from chicks fed 15 mg Zn/kg from ZnSO4. Weight gain was not different (P > 0.05) when comparing the 2 sources of Zn hydroxychloride supplemented at 10 mg Zn/kg. Tibia ash Zn and total tibia Zn were increased (P < 0.05) by all Zn sources and responded linearly (P < 0.05) to Zn supplementation from ZnSO4 and SAMZn. Total tibia Zn concentration was not different (P > 0.05) for chicks fed 10 mg Zn/kg from either source of Zn hydroxychloride. Multiple linear regression of total tibia Zn on supplemental Zn intake (R2 = 0.95) resulted in a RBV of 115% for SAMZn compared with ZnSO4 (set at 100%). The RBV of SAMZn was higher (P < 0.05) than ZnSO4. In conclusion, relative bioavailability of Zn (based on tibia Zn) in Zn hydroxychloride from SAMZn was higher than feed grade ZnSO4 based on multiple regression slope-ratio analysis and was similar to that of IBZn Zn hydroxychloride based on tibia Zn responses to 10 mg/kg supplemental dietary Zn.

Determination of TMEn, standardized amino acid digestibility, phosphorus digestibility, and phosphorus bioavailability in conventional and reduced phosphorus distillers dried grains with solubles.

The TMEn, amino acid (AA) digestibility, and P availability in 2 conventional corn distillers dried grains with solubles (C-DDGS1 and 2; 0.86 to 1.14% P, DM basis) and reduced phosphorus DDGS (RP-DDGS; 0.39% P) were evaluated. The TMEn of C-DDGS1 and 2 and RP-DDGS were determined in Experiment 1 using conventional adult Leghorn roosters, while standardized AA digestibility was determined in Experiment 2 using cecectomized roosters. Apparent ileal digestibility (AID) of P at different Ca levels was determined using precision-fed (crop intubation) broiler chickens in Experiments 3 and 4. The AID and total tract retention of P in C-DDGS2 were evaluated in Experiment 5 using ad libitum-fed broilers. Phosphorus bioavailability in C-DDGS2 relative to KH2PO4 based on bone ash was determined in Experiment 6. Experiments contained 4 to 5 replicates per treatment. In Experiment 1, the TMEn of C-DDGS1 and RP-DDGS was 3,428 and 2,840 kcal/kg, respectively (DM basis). In Experiment 2, there were no differences (P > 0.05) in rooster AA digestibility values between C-DDGS1 and RP-DDGS. In Experiment 3 with precision-fed chicks, AID of P in C-DDGS1 and RP-DDGS was 81 and 59%, respectively; there was no effect (P > 0.05) of increasing dietary Ca level from 0.04 to 1.0% for C-DDGS1 or reducing Ca from 1.5 to 1.0% for RP-DDGS. The AID of P in precision-fed chicks for C-DDGS2 in Experiment 4 was 48 and 80% at 1.3 and 0.3 Ca:total P ratios, respectively (P < 0.05). In Experiment 5, AID of P in C-DDGS2 at Ca:total P ratios of 1.3 and 2.5 was 63 and 42%, respectively, in precision-fed chicks. Regression of bone ash content (mg/tibia) on supplemental P intake in Experiment 6 yielded a P bioavailability of 61% relative to KH2PO4 for C-DDGS2. In conclusion, total and digestible P content in RP-DDGS was greatly reduced compared with C-DDGS, and the digestibility and bioavailability of the P in C-DDGS was affected by type of experimental assay and dietary Ca level.

Quantification of secretory IgA and mucin excretion and their contributions to total endogenous amino acid losses in roosters that were fasted or precision-fed a nitrogen-free diet or various highly digestible protein sources.

The objective of this study was to quantify total secretory IgA (sIgA) and mucin excretion via excreta in roosters fed diets containing highly digestible protein sources and to determine their proportional contributions to total endogenous amino acid (AA) losses. Precision-fed rooster assays with 24 h excreta collections were conducted using conventional White Leghorn roosters (4-8 roosters per treatment). In Experiment 1, roosters were fasted or precision-fed 30 g (crop intubation) of a nitrogen-free (NF) or semi-purified diet containing 10% casein. Roosters in Experiment 2 received a NF or semi-purified diet containing either 10% casein, 17% whole egg, 10% egg white, 9.8% soy protein isolate, 10.2% chicken breast meat, 11.2% spray-dried animal plasma (SDAP), or an AA mixture containing the same AA as casein. A Latin square design was used in Experiment 3, where roosters received NF or semi-purified diets containing either 10% casein, 17% whole egg, or 9.6% of a crystalline AA mixture to evaluate both diet and individual bird effects. In Experiment 1, mucin excretion did not differ (P > 0.05) among treatments; however, total sIgA excretion was lower for fasted birds, intermediate for the NF diet, and highest for casein (P < 0.05). Total endogenous AA losses (proportion of the total) from sIgA were higher for roosters fed casein, whereas mucin contributions were higher for fasted roosters (P < 0.05). In Experiment 2, sIgA excretion did not differ (P > 0.05) among treatments; however, mucin excretion was reduced for NF, whole egg, egg white, and chicken breast compared with casein and SDAP. In Experiment 3, sIgA and mucin excretion did not differ (P > 0.05) among treatments; however, sIgA excretion differed among individual roosters (7-27 mg/24 h; P < 0.05). Overall, fasting reduced sIgA excretion and sIgA and mucin excretion were affected by dietary protein source. Further, roosters excreted a substantial amount of sIgA, and sIgA and mucin were considerable contributors to total endogenous AA losses.

Standardized amino acid digestibility and true metabolizable energy for several increased protein ethanol co-products produced using back-end fractionation systems.

Precision-fed rooster trials were conducted to evaluate standardized AA digestibility and TMEn of the increased protein ethanol co-products corn fermented protein (CFP), high protein-distillers dried grains with solubles (HP-DDGS), and reduced fiber high protein-DDGS (RFHP-DDGS) produced using post-fermentation back-end fractionation systems. The TMEn was determined using conventional adult Leghorn roosters, while cecectomized roosters were used to determine standardized AA digestibility. Three to 6 roosters were fasted per treatment for 26 h prior to crop intubation with 27 g of sample and excreta were collected for 48 h post-feeding. Statistical analyses were conducted using a one-way ANOVA for a completely randomized design. Eight samples of CFP were found to contain a mean of 56% CP (DM basis) compared with a mean of 32% for conventional DDGS. The mean TMEn of CFP (3,556 kcal/kg) was greater (P < 0.05) than conventional DDGS1 and 2 (2,767 kcal/kg DM); mean standardized AA digestibility for CFP was similar to conventional DDGS and ranged from 88 to 94%. The mean digestible Lys, Met+Cys, and Thr concentrations for conventional DDGS were 0.79, 1.12, and 0.94%, respectively, whereas those for CFP were 1.74, 2.06, and 1.88%, respectively (DM basis). Two samples of HP-DDGS contained a mean of 51% CP (DM basis), a mean TMEn of 3,325 kcal/kg DM, a mean standardized AA digestibility of 90%, and mean concentrations of digestible Lys, Met+Cys, and Thr, which were 1.53, 1.77, and 1.60%, respectively (DM basis). The mean CP content of 2 RFHP-DDGS was 48% and the mean TMEn was 3,711 kcal/kg DM, which was greater (P < 0.05) than conventional DDGS3 and 4 (2,920 kcal/kg DM). Mean standardized AA digestibility of RFHP-DDGS was 90% and mean digestible Lys, Met+Cys, and Thr concentrations increased from 0.82, 1.01, and 0.95% for conventional DDGS, respectively, to 1.00, 1.59, and 1.44% for RFHP-DDGS, respectively (DM basis). Results indicate these high protein corn ethanol co-products have increased nutritional value for poultry compared with conventional DDGS.

Research Note: Evaluation of a precision-fed rooster assay for determination of phytic acid disappearance in feedstuffs.

The objective of this study was to evaluate a precision-fed rooster assay that is suitable for determination of phytic acid (myo-inositol 1,2,3,4,5,6-hexakis; InsP6) disappearance in plant-based feed ingredients. A 48-h precision-fed rooster assay was used to measure InsP6 disappearance using conventional White Leghorn roosters. A minimum of 4 individually-caged roosters per treatment were fasted for 26 h prior to crop intubation with 15 to 30 g of sample, and excreta were quantitatively collected for 48 h. Soybean meal, soybean hulls, canola meal, conventional distillers dried grains with solubles (DDGS), palm kernel meal (PKM), and wheat bran were evaluated in Experiment 1, whereas wheat middlings (WM) and rice bran (RB) were evaluated without and with 1,000 and 1,800 U/kg phytase in Experiment 2. Data from Experiment 1 were subjected to a one-way ANOVA for a completely randomized design, while data from Experiment 2 were subjected to two-way ANOVA for a 2 × 3 factorial arrangement of treatments. In Experiment 1, InsP6 disappearance ranged from 3 to 95% among all ingredients. The InsP6 disappearance for conventional DDGS (95%) was the highest (P < 0.05), wheat bran and soybean hulls were intermediate (47-48%), PKM was low (24%), and soybean meal and canola meal were very low (3-5%). In Experiment 2, there was a significant ingredient × phytase interaction (P < 0.05). Phytase inclusion at both 1,000 and 1,800 U/kg resulted in a significant improvement (P < 0.05) in InsP6 disappearance for RB; however, only the addition of 1,800 U/kg resulted in an increase in InsP6 disappearance for WM. The addition of 1,800 U/kg phytase increased the InsP6 disappearance from 58 to 74% for WM and from 26 to 53% for RB. These results suggest the precision-fed rooster assay can be used to evaluate phytic acid disappearance in plant-based feed ingredients and the assay was able to detect a significant effect of 1,800 U/kg of exogenous phytase on phytic acid disappearance for WM and RB.

Metabolizable energy and amino acid digestibility in spray-dried animal plasma using broiler chick and precision-fed rooster assays.

Four experiments were conducted to determine ME and amino acid (AA) digestibility of spray-dried animal plasma (SDAP) and soybean meal (SBM). The 48-h precision-fed adult rooster assay was used in 2 experiments; TMEn and standardized AA digestibility were determined using conventional and cecectomized roosters, respectively, 50 weeks of age and weighing approximately 2,200 g. Eight individually-caged roosters (4 per diet) were fasted for 26 h, then precision-fed 30 g of SDAP mixture (containing 50% corn) or SBM mixture (containing 50% corn). The TMEn and AA digestibility for SDAP and SBM were calculated by the difference procedure. The TMEn for SDAP was greater (P < 0.05) than SBM (3,743 and 2,669 kcal/kg DM, respectively). Similarly, mean AA digestibility of SDAP was greater (P < 0.05) than SBM (94 and 86%, respectively). Two assays were conducted using Ross male broilers to determine AMEn and apparent (AIAAD) and standardized (SIAAD) ileal AA digestibility of SDAP and SBM. A 3 × 2 factorial arrangement of treatments was used to determine AMEn; 126 chicks (6 replicate pens of 7 chicks) were fed a corn-SBM-based reference diet, a diet containing 30% SDAP, or a diet containing 30% SBM from d 7 to 10 and 18 to 21. A 2 × 2 factorial arrangement of treatments was used to determine AIAAD and SIAAD; 168 chicks (12 replicate pens of 7 chicks) were fed a semi-purified diet containing 25% SDAP or a semi-purified, isonitrogenous diet containing 41% SBM from d 7 to 10 and 18 to 21. The AMEn for SDAP was greater (P < 0.05) than SBM at d 10 (3,851 and 2,089 kcal/kg DM, respectively) and d 21 (4,239 and 2,849 kcal/kg DM, respectively). The second assay showed an increase (P < 0.05) in AIAAD and SIAAD for SDAP compared with SBM at d 10 (mean SIAAD for SDAP and SBM were 96% and 84%, respectively) and d 21 (97% and 87%, respectively). Regardless of assay or age, these results indicate SDAP is a highly digestible feed ingredient with high ME and AA digestibility.

Relationship between enteric pathogens and acute gastroenteritis disease severity: a prospective cohort study.

OBJECTIVES: To evaluate the relationship between individual bacterial and viral pathogens and disease severity. METHODS: Children <18 years with three or more episodes of vomiting and/or diarrhoea were enrolled in two Canadian paediatric emergency departments between December 2014 and August 2016. Specimens were analysed employing molecular panels, and outcome data were collected 14 days after enrolment. The primary outcome was severe disease over the entire illness (symptom onset until 14-day follow-up), quantified employing the Modified Vesikari Scale (MVS) score. The score was additionally analysed in two other time periods: index (symptom onset until enrolment) and follow-up (enrolment until 14-day follow-up). RESULTS: Median participant age was 20.7 (IQR: 11.3, 44.2) months; 47.4% (518/1093) and 73.4% (802/1093) of participants had index and total MVS scores ≥11, respectively. The most commonly identified pathogens were rotavirus (289/1093; 26.4%) and norovirus (258/1093; 23.6%). In multivariable analysis, severe disease over the entire illness was associated with rotavirus (OR = 9.60; 95%CI: 5.69, 16.19), Salmonella (OR = 6.61; 95%CI: 1.50, 29.17), adenovirus (OR = 2.53; 95%CI: 1.62, 3.97), and norovirus (OR = 1.43; 95%CI: 1.01, 2.01). Pathogens associated with severe disease at the index visit were: rotavirus only (OR = 6.13; 95%CI: 4.29, 8.75), Salmonella (OR = 4.59; 95%CI: 1.71, 12.29), adenovirus only (OR = 2.06; 95%CI: 1.41, 3.00), rotavirus plus adenovirus (OR = 3.15; 95%CI: 1.35, 7.37), and norovirus (OR = 0.68; 95%CI: 0.49, 0.94). During the follow-up period, rotavirus (OR = 2.21; 95%CI: 1.50, 3.25) and adenovirus (OR = 2.10; 95%CI: 1.39, 3.18) were associated with severe disease. CONCLUSIONS: In children presenting for emergency department care with acute gastroenteritis, pathogens identified were predominantly viruses, and several of which were associated with severe disease. Salmonella was the sole bacterium independently associated with severe disease.

The efficacy of pregabalin for treating pain associated with diabetic peripheral neuropathy in subjects with type 1 or type 2 diabetes mellitus.

OBJECTIVE: The objective of this study was to compare the efficacy and safety of pregabalin for painful diabetic peripheral neuropathy (pDPN) in subjects with type 1 (T1DM) or 2 diabetes mellitus (T2DM). METHODS: Pooled data from 10 randomized clinical trials (pregabalin-treated T1DM and T2DM subjects with pDPN) were analyzed for change from baseline (CFB) scores (pain and sleep disturbance) using mixed model repeated measures (MMRM) through Week 12 and last observation carried forward (LOCF). Adverse events (AEs) were recorded. RESULTS: Pregabalin-treated (T1DM 156 [8.7%]; T2DM 1632 [91.3%]) and placebo subjects (T1DM 92 [9.6%]; T2DM 868 [90.4%]) had comparable baseline demographic characteristics between treatment groups within the same diabetes type. T2DM (vs. T1DM) subjects were ∼10 years older. With pregabalin and placebo, respectively, mean ± SD baseline pain (T1DM: 6.2 ± 1.4 and 6.5 ± 1.6; T2DM: 6.5 ± 1.5 and 6.4 ± 1.5) and sleep scores (T1DM: 5.2 ± 2.4 and 5.2 ± 2.7; T2DM: 5.3 ± 2.5 and 5.1 ± 2.5) were comparable. Using MMRM, mean CFB treatment differences (pregabalin minus placebo) were significantly different for pain and sleep with either diabetes types (all weeks p < .05). With LOCF, pregabalin's odds ratios (ORs) of achieving 30% pain reduction were similar with T2DM (OR, 1.91, 95% CI [1.61, 2.27]) and T1DM (2.01 [1.18, 3.44]) (both p ≤ .01). Pregabalin's ORs of 30% improvement in sleep quality were 1.81 (95% CI, 1.06, 3.09) with T1DM and 2.01 (1.69, 2.39) with T2DM (both p < .05). AEs were consistent with the known safety profile of pregabalin. CONCLUSIONS: Pregabalin significantly improved pain and sleep quality, without a clinically meaningful difference between diabetes types. ClinicalTrial.gov registration: NCT00156078, NCT00159679, NCT00143156, NCT00553475.

Loss of PRDM1/BLIMP-1 function contributes to poor prognosis of activated B-cell-like diffuse large B-cell lymphoma.

PRDM1/BLIMP-1, a master regulator of plasma-cell differentiation, is frequently inactivated in activated B-cell-like (ABC) diffuse large B-cell lymphoma (DLBCL) patients. Little is known about its genetic aberrations and relevant clinical implications. A large series of patients with de novo DLBCL was effectively evaluated for PRDM1/BLIMP-1 deletion, mutation, and protein expression. BLIMP-1 expression was frequently associated with the ABC phenotype and plasmablastic morphologic subtype of DLBCL, yet 63% of the ABC-DLBCL patients were negative for BLIMP-1 protein expression. In these patients, loss of BLIMP-1 was associated with Myc overexpression and decreased expression of p53 pathway molecules. In addition, homozygous PRDM1 deletions and PRDM1 mutations within exons 1 and 2, which encode for domains crucial for transcriptional repression, were found to show a poor prognostic impact in patients with ABC-DLBCL but not in those with germinal center B-cell-like DLBCL (GCB-DLBCL). Gene expression profiling revealed that loss of PRDM1/BLIMP-1 expression correlated with a decreased plasma-cell differentiation signature and upregulation of genes involved in B-cell receptor signaling and tumor-cell proliferation. In conclusion, these results provide novel clinical and biological insight into the tumor-suppressive role of PRDM1/BLIMP-1 in ABC-DLBCL patients and suggest that loss of PRDM1/BLIMP-1 function contributes to the overall poor prognosis of ABC-DLBCL patients.

An evaluation of fluid immersion therapy for the prevention of pressure ulcers.

BACKGROUND: Individuals with impaired mobility can spend prolonged periods on support surfaces, increasing their risk of developing pressure ulcers. Manufacturers have developed mattresses to maximise contact area. The present study evaluated both the biomechanical and physiological responses to lying postures on a Fluid Immersion Simulation mattress. METHODS: Seventeen healthy participants were recruited to evaluate the mattress during three prescribed settings of immersion (high, medium and low). Parameters reflecting biomechanical and physiological responses, and the microclimate were monitored during three postures (supine, lateral and high-sitting) over a 90minute test session. Transcutaneous oxygen and carbon dioxide gas responses were categorised according to three criteria and data were compared between each condition. FINDINGS: Results indicated that interface pressures remained consistent, with peak sacral values ranging from 21 to 27mmHg across all immersion settings and postures. The majority of participants (82%) exhibited minimal changes in gas tensions at the sacrum during all test conditions. By contrast, three participants exhibited decreased oxygen with increased carbon dioxide tensions for all three immersion settings. Supine and high sitting sacral microclimate values ranged between 30.1-30.6°C and 42.3-44.5% for temperature and relative humidity respectively. During lateral tilt there was a reduction of 1.7-2.5°C and 3.3-5.3% in these values. The majority of participants reported high comfort scores, although a few experienced bottoming out during the high-sitting posture at the high immersion setting. INTERPRETATION: Fluid Immersion Simulation provides an intelligent approach to increase the support area. Further research is required to provide evidence based guidance on the use of personalised support surfaces.

Primary testicular diffuse large B-cell lymphoma displays distinct clinical and biological features for treatment failure in rituximab era: a report from the International PTL Consortium.

Primary testicular diffuse large B-cell lymphoma (PT-DLBCL) is a unique subtype of DLBCL. The impact of rituximab on survival and patterns of treatment failure in PT-DLBCL patient remain controversial. We analyzed the clinical and biological feature of 280 PT-DLBCL cases, 64% of which were treated with rituximab-containing regimens. Although most (95%) patients achieved complete remission, a continuous risk of relapse was observed. Rituximab significantly reduced the cumulative risk of relapse (P=0.022) and improved both progression-free survival (PFS, P=0.012) and overall survival (OS, P=0.027) of PT-DLBCL patients (5-year PFS, 56% vs 36%; 5-year OS, 68% vs 48%). Central nervous system and contralateral testis were the most common sites of relapse, but other extranodal and nodal sites of relapse were also observed. Most cases of PT-DLBCL had a non-germinal center B-cell like (84%) immunophenotype and an activated B-cell like (86%) gene expression profile (GEP) subtype. The distinctive GEP signature of primary testicular lymphoma was relevant to tumor cell proliferation, dysregulated expression of adhesion molecules and immune response, likely accounting for the poor outcome. Accordingly, forkhead box P1 transcription factor (FOXP1) and T-cell leukemia/lymphoma 1 (TCL1) oncogenic activation were confirmed and predicted a significant trend of poor survival. This study provides valuable observations for better understanding of both clinical and biological features in PT-DLBCL patients.

The Hippo pathway promotes cell survival in response to chemical stress.

Cellular stress defense mechanisms have evolved to maintain homeostasis in response to a broad variety of environmental challenges. Stress signaling pathways activate multiple cellular programs that range from the activation of survival pathways to the initiation of cell death when cells are damaged beyond repair. To identify novel players acting in stress response pathways, we conducted a cell culture RNA interference (RNAi) screen using caffeine as a xenobiotic stress-inducing agent, as this compound is a well-established inducer of detoxification response pathways. Specifically, we examined how caffeine affects cell survival when Drosophila kinases and phosphatases were depleted via RNAi. Using this approach, we identified and validated 10 kinases and 4 phosphatases that are essential for cell survival under caffeine-induced stress both in cell culture and living flies. Remarkably, our screen yielded an enrichment of Hippo pathway components, indicating that this pathway regulates cellular stress responses. Indeed, we show that the Hippo pathway acts as a potent repressor of stress-induced cell death. Further, we demonstrate that Hippo activation is necessary to inhibit a pro-apoptotic program triggered by the interaction of the transcriptional co-activator Yki with the transcription factor p53 in response to a range of stress stimuli. Our in vitro and in vivo loss-of-function data therefore implicate Hippo signaling in the transduction of cellular survival signals in response to chemical stress.

Oxidation of glycosaminoglycans by free radicals and reactive oxidative species: A review of investigative methods.

Glycosaminoglycans, in particular hyaluronan (HA), and proteoglycans are components of the extracellular matrix (ECM). The ECM plays a key role in the regulation of cellular behaviour and alterations to it can modulate both the development of human diseases as well as controlling normal biochemical processes such as cell signalling and pro-inflammatory responses. For these reasons, in vitro fragmentation studies of glycosaminoglycans by free radicals and oxidative species are seen to be relevant to the understanding of in vivo studies of damage to the ECM. A wide range of investigative techniques have therefore been applied to gain insights into the relative fragmentation effects of several reactive oxidative species with the ultimate goal of determining mechanisms of fragmentation at the molecular level. These methods are reviewed here.

Critique of the analysis of UpToDate.com on the treatment of painful vertebral compression fractures: time to update UpToDate.

The treatment of painful vertebral compression fractures has changed substantially since the introduction of vertebroplasty in the mid-1980s and balloon kyphoplasty in the late 1990s. Both procedures were widely accepted with the vertebral fractures treated reaching 150,000 per annum in 2009 prior to the publication of 2 randomized controlled trials comparing vertebroplasty with a sham treatment published in the New England Journal of Medicine in August 2009. Since then, there has been a flood of information on vertebral augmentation and balloon kyphoplasty. It is worth evaluating this information especially because it relates to current recommendations that are often followed blindly by medical and administrative groups unfamiliar with either the procedure or the high level of evidence surrounding vertebral augmentation. To streamline the evaluation of some current recommendations, we limited the analysis to the recommendations found on UpToDate.com. This Web site is an evidence-based, peer-reviewed source of information available for patients, doctors, health insurance companies, and population-based medical decision-making.

Soluble plantain nonstarch polysaccharides, although increasing caecal load, reduce systemic invasion of Salmonella Gallinarum in the chicken.

UNLABELLED: Soluble plantain (Musa paradisiaca) nonstarch polysaccharides (NSPs) have previously been shown to prevent pathogenic interaction with the intestinal epithelium. Here, we examined whether plantain NSP could prevent the invasion of the intestinal mucosa by Salmonella enterica Gallinarum, a causative agent of fowl typhoid. In vitro assays using B1OXI cells were performed with monolayers pretreated with/without plantain NSP, before inoculation with Salm. Gallinarum 287/91. Chicks were fed from hatch on a pellet diet containing 50 mg day(-1) plantain NSP, followed by oral inoculation with Salm. Gallinarum 287/91 at the age of 6 days. Bacteria were enumerated from the liver, spleen and caecal contents 3 days postinfection. Adhesion and invasion of Salm. Gallinarum to B1OXI cells were inhibited by 10 mg ml(-1) plantain NSP (reduction in invasion 52%; 95% CI 27-77; P < 0.05). In vivo diet supplemented with 50 mg day(-1) plantain NSP reduced the invasion of Salm. Gallinarum in the chick liver (control 4.05 Log10 CFU g(-1) , SE 0.28, vs plantain 2.07 Log10 CFU g(-1) , SE 0.65; P < 0.01) and nonsignificantly in the spleen. Conversely, CFUs were significantly increased in the caeca (control 1.27 Log10 CFU g(-1), SE 0.65, vs plantain 6.04 Log10 CFU g(-1) , SE 0.17; P < 0.001). Plantain NSP feed reduced the systemic infection by Salm. Gallinarum and may have potential in reducing the impact of the disease in avian salmonellosis. The caveat is the increased caecal load of Salm. Gallinarum, although this may reflect both the reduced intestinal invasion and the bacterial multiplication in the caeca. SIGNIFICANCE AND IMPACT OF THE STUDY: Nonstarch polysaccharide (NSP) derived from the plantain (Musa paradisiaca) inhibits interaction with epithelial cells by Salmonella enterica Gallinarum, a causative agent of the disease fowl typhoid. Incorporation of plantain NSP into the poultry feed reduced Salm. Gallinarum levels in the spleen and liver of chicks following experimental infection, although their numbers in the caeca increased. These data demonstrate that alternatives to antimicrobials such as NSP may be useful in the control of invasive salmonellosis in poultry.

Infection of chickens with antimicrobial-resistant Salmonella enterica Typhimurium DT193 and monophasic Salmonella Typhimurium-like variants: an emerging risk to the poultry industry?

Antimicrobial-resistant Salmonella enterica poses a particular risk to public health, and in particular isolates belonging to clonal lineages such as Salmonella Typhimurium DT104 cause epidemics across species including poultry. In recent years, antimicrobial-resistant S. Typhimurium DT193 and specifically the monophasic S. Typhimurium-like variants of this phage type, serotypes 4,12:i:- and 4,5,12:i:-, have become an increasing risk to public health in Europe and the USA and now account for nearly one-half of human S. Typhimurium infections in the UK. Unlike S. Typhimurium that possesses two forms of flagella which can vary between phase 1 and phase 2 during infection, monophasic variants possess only phase 1 flagella. These monophasic antimicrobial-resistant variants have become a major problem in pig production but human cases have also been associated with poultry consumption and have been found in UK flocks through surveillance schemes since 2010. In this study we determined the ability of antimicrobial-resistant DT193 serotype 4,12:i:- and 4,5,12:i:- isolates from pigs to infect chickens. All isolates were found to colonize the caeca and liver. All but one isolate of serotype 4,5,12:i:- also infected the spleen. Levels of infection and pathology were comparable with those found with the virulent S. Typhimurium isolate 4/74. These findings indicate that both S. Typhimurium DT193 and monophasic variants of this phage type usually associated with pigs are capable of colonizing the chicken. This shows that both S. Typhimurium DT193 and monophasic variants represent a significant and potential emerging threat to poultry production from "spill-over" of these isolates from the pig industry or other sources.

One-electron oxidation and reduction of glycosaminoglycan chloramides: a kinetic study.

Hypochlorous acid and its acid-base counterpart, hypochlorite ions, produced under inflammatory conditions, may produce chloramides of glycosaminoglycans, these being significant components of the extracellular matrix (ECM). This may occur through the binding of myeloperoxidase directly to the glycosaminoglycans. The N-Cl group in the chloramides is a potential selective target for both reducing and oxidizing radicals, leading possibly to more efficient and damaging fragmentation of these biopolymers relative to the parent glycosaminoglycans. In this study, the fast reaction techniques of pulse radiolysis and nanosecond laser flash photolysis have been used to generate both oxidizing and reducing radicals to react with the chloramides of hyaluronan (HACl) and heparin (HepCl). The strong reducing formate radicals and hydrated electrons were found to react rapidly with both HACl and HepCl with rate constants of 1-1.7 × 10(8) and 0.7-1.2 × 10(8)M(-1)s(-1) for formate radicals and 2.2 × 10(9) and 7.2 × 10(8)M(-1)s(-1) for hydrated electrons, respectively. The spectral characteristics of the products of these reactions were identical and were consistent with initial attack at the N-Cl groups, followed by elimination of chloride ions to produce nitrogen-centered radicals, which rearrange subsequently and rapidly to produce C-2 radicals on the glucosamine moiety, supporting an earlier EPR study by M.D. Rees et al. (J. Am. Chem. Soc.125: 13719-13733; 2003). The oxidizing hydroxyl radicals also reacted rapidly with HACl and HepCl with rate constants of 2.2 × 10(8) and 1.6 × 10(8)M(-1)s(-1), with no evidence from these data for any degree of selective attack on the N-Cl group relative to the N-H groups and other sites of attack. The carbonate anion radicals were much slower with HACl and HepCl than hydroxyl radicals (1.0 × 10(5) and 8.0 × 10(4)M(-1)s(-1), respectively) but significantly faster than with the parent molecules (3.5 × 10(4) and 5.0 × 10(4)M(-1)s(-1), respectively). These findings suggest that these potential in vivo radicals may react in a site-specific manner with the N-Cl group in the glycosaminoglycan chloramides of the ECM, possibly to produce more efficient fragmentation. This is the first study therefore to conclusively demonstrate that reducing radicals react rapidly with glycosaminoglycan chloramides in a site-specific attack at the N-Cl group, probably to produce a 100% efficient biopolymer fragmentation process. Although less reactive, carbonate radicals, which may be produced in vivo via reactions of peroxynitrite with serum levels of carbon dioxide, also appear to react in a highly site-specific manner at the N-Cl group. It is not yet known if such site-specific attacks by this important in vivo species lead to a more efficient fragmentation of the biopolymers than would be expected for attack by the stronger oxidizing species, the hydroxyl radical. It is clear, however, that the N-Cl group formed under inflammatory conditions in the extracellular matrix does present a more likely target for both reactive oxygen species and reducing species than the N-H groups in the parent glycosaminoglycans.