Distillers dried grains with soluble and enzyme inclusion in the diet effects broilers performance, intestinal health, and microbiota composition.

This study tested the effect of distillers dried grains with soluble (DDGS) inclusion in a broiler diet, with or without supplementation of exogenous enzymes, on the microbiota composition, intestinal health, diet digestibility and performance. A total of 288 one-day-old chickens was assigned to 6 treatments (8 replicate of 6 birds each) according to a completely randomized design with a 3 × 2 factorial scheme with 3 DDGS levels (0, 7 and 14%) and 2 inclusions of exogenous enzymes (with or without a multicarbohydrase complex + phytase [MCPC]). The results exhibited that DDGS inclusion up to 14% did not impair broilers performance up to 28 d, however, DDGS-fed animals exhibited significant improvement with the MCPC supplementation. No effects of the enzymes in the ileal digestibility were found at 21 d. DDGS inclusion in the diet affected dry matter and gross energy digestibility. Broilers fed diets with MCPC were found to have less intestinal histological alteration thus better gut health. No effect of DDGS, enzyme or interaction of those were observed for intestinal permeability and in the serum inflammatory biomarker (calprotectin) at 7 and 28 d. The increase of DDGS percentage in the diet reduced the diversity of the ileal microbiota but increased the cecal microbiota diversity. The inclusion of DDGS showed positive effects on microbiota composition due to a reduction of Proteobacteria phylum in the ileum at 28d and a reduction in the presence of Enterococcaceae family in the ileum at 14 and 28d. The inclusion of MCPC complex might promote beneficial changes in the ileal and cecal microbiota due reduce of Proteobacteria, Bacillaceae and Enterobacteriaceae. The supplementation of xylanase, ß-glucanase, arabinofuranosidase and phytase to a DDGS diet improves performance and intestinal health allowing the use of these subproduct in the poultry nutrition.

Evaluation of biofilm formation using milk in a flow cell model and microarray characterization of Staphylococcus aureus strains from bovine mastitis.

It was hypothesized that biofilm could play an important role in the establishment of chronic Staphylococcus aureus bovine mastitis. The in vitro evaluation of biofilm formation can be performed either in closed/static or in flow-based systems. Efforts have been made to characterize the biofilm-forming ability of S. aureus mastitis isolates, however most authors used static systems and matrices other than UHT milk. It is not clear whether such results could be extrapolated to the mammary gland environment. Therefore, the present study aimed to investigate the biofilm-forming ability of S. aureus strains from subclinical bovine mastitis using the static method and a flow-based one. One hundred and twelve strains were tested by the classic tissue culture plate assay (TCP) and 30 out of them were also tested by a dynamic semi-quantitative assay using commercial UHT milk as culture medium (Milk Flow Culture, MFC) or Tryptic Soy Broth as control medium (TS Flow Culture, TSFC). Only 6 (20%) strains formed biofilm in milk under flow conditions, while 36.6% were considered biofilm-producers in TCP, and 93.3% produced biofilm in TSFC. No agreement was found between TCP, MFC and TSFC results. The association between strain genetic profile, determined by microarray, and biofilm-forming ability in milk was evaluated. Biofilm formation in MFC was significantly associated with the presence of those genes commonly found in bovine-associated strains, assigned to clonal complexes typically detected in mastitis. Based on our results, biofilm-forming potential of bovine strains should be critically analysed and tested applying conditions similar to mammary environment.

High doses of pseudoephedrine hydrochloride accelerate onset of CNS oxygen toxicity seizures in unanesthetized rats.

Pseudoephedrine (PSE) salts (hydrochloride and sulfate) are commonly used as nasal and paranasal decongestants by scuba divers. Anecdotal reports from the Divers Alert Network suggest that taking PSE prior to diving while breathing pure O2 increases the risk for CNS oxygen toxicity (CNS-OT), which manifests as seizures. We hypothesized that high doses of PSE reduce the latency time to seizure (LS) in unanesthetized rats breathing 5 atmospheres absolute (ATA) of hyperbaric oxygen. Sixty-three male rats were implanted with radio-transmitters that recorded electroencephalogram activity and body temperature. After ≥7-day recovery, and 2 h before "diving", each rat was administered either saline solution (control) or PSE hydrochloride intragastrically at the following doses (mg PSE/kg): 0, 40, 80, 100, 120, 160, and 320. Rats breathed pure O2 and were dived to 5ATA until the onset of behavioral seizures coincident with neurological seizures. LS was the time elapsed between reaching 5ATA and exhibiting seizures. We observed a significant dose-dependent decrease in the LS at doses of 100-320 mg/kg, whereas no significant differences in LS from control value were observed at doses ≤80 mg/kg. Our findings showed that high doses of PSE accelerate the onset of CNS-OT seizures in unanesthetized rats breathing 5ATA of poikilocapnic hyperoxia. Extrapolating our findings to humans, we conclude that the recommended daily dose of PSE should not be abused prior to diving with oxygen-enriched gas mixes or pure O2.

Differential cell count as an alternative method to diagnose dairy cow mastitis.

Changes in relative cell proportions occurring in diseased mammary glands of dairy cows can be determined using differential cell count (DCC). The present study was carried out in 2 consecutive trials, with 2 goals: (a) to test the consistency of DCC results on subsequent days, and (b) to establish an effective cutoff value for the diagnosis of mastitis. In the first trial, quarter milk and blood samples were taken from 8 healthy cows for 5 consecutive days. Milk samples were tested by somatic cell count (SCC) and bacteriological analysis, and DCC was performed on blood and milk samples by flow cytometer. In the second trial, 16 animals were randomly selected from a different herd and quarter milk samples taken on 3 consecutive milkings. All samples were cyto-bacteriologically analyzed and DCC was performed on the second sampling. In the first trial, mean SCC was 77,770 cells/mL and 4 samples were bacteriologically positive. No fixed or random effect had a significant influence on percentages of individual cell populations or ratios in blood or milk. A cutoff value of 0.495 for logarithmic polymorphonuclear neutrophilic leukocyte:lymphocyte ratio was established. Mean SCC of milk samples collected in the second trial was 543,230 cells/mL, and infection was detected in 53.1% of quarters, mostly caused by Staphylococcus aureus. When the cutoff value was applied to the data along with SCC, sensitivity and specificity of the diagnostic method were 97.3 and 92.3%, respectively.

Microscopic differential cell counting to identify inflammatory reactions in dairy cow quarter milk samples.

The diagnosis of intramammary infections is mostly based on somatic cell count (SCC) and bacteriological analysis. As an alternative, differential cell counting (DCC) could be a useful method, because it identifies changes in the relative cell populations before the increase in total cell number occurs. The aim of the study was to identify cytological parameters that could be used in the field to classify mammary quarters as healthy or diseased, comparing cyto-bacteriological results with DCC. Overall, 48 cows were randomly selected from 3 herds in Lombardy region of Italy. Herd A was characterized by the absence of contagious microorganisms; in herds B and C, the prevalence of Staphylococcus aureus was 20 and 50%, respectively. Foremilk samples were aseptically collected from 188 quarters and submitted to bacteriological analysis, SCC, and DCC. For statistical analysis, the samples were clustered into 4 health groups, and DCC results were compared in each group. Ninety-six samples were classified as normal secretions (N), 30 as mastitis (M), 15 as latent mastitis (LM), and 47 as unspecific mastitis (UM) based on SCC and bacteriological results. Single percentages of lymphocytes, polymorphonuclear neutrophilic leukocytes (PMNL), or macrophages were first evaluated to established variables capable of identifying healthy and inflamed quarters. Then, combinations of cell populations were tested to increase the discrimination power of DCC: phagocytes, logarithmic PMNL:lymphocyte ratio, and logarithmic phagocyte:lymphocyte ratio. The mean percentage of lymphocytes was significantly higher in group N than in groups LM, UM, and M. The mean percentage of PMNL was significantly lower in group N than in groups UM and M, but not LM. Mean percentages of macrophages were not significantly influenced by the 4 groups. The mean value of phagocytes was significantly lower in group N than in the other groups. Both the logarithmic PMNL:lymphocyte and the logarithmic phagocyte:lymphocyte ratios were significantly lower in group N than in groups LM, UM, and M. Fisher (F-)values were calculated, and the highest F-value was that of log PMNL:lymphocytes ratio (48.23). The explanation for this could be that log PMNL:Lym is the only variable that involved both cell populations statistically influenced by health groups but excluded macrophages. Microscopic DCC has potential as a tool to identify cows affected by any inflammatory process of the mammary gland, with the best results being achieved using log PMNL:lymphocyte as variable.

Effect of pre-breathing oxygen at different depth on oxidative status and calcium concentration in lymphocytes of scuba divers.

AIM: In-water pre-breathing oxygen at various depths reduces decompression-induced bubble formation and platelet activation, but it could induce side effects such as oxidative stress. The aim of this study was to investigate the effect of in-water pre-breathing oxygen, at different depths, on the oxidative status and intracellular calcium ([Ca(2+) ]i) of peripheral blood lymphocytes isolated from six divers. They participated in a 4-diving protocol. Two week recovery time was allowed between successive dives. Before diving, all divers, for 20 min, breathed normally at sea level (dive 1), 100% oxygen at sea level (dive 2), 100% oxygen at 6 msw (dive 3), 100% oxygen at 12 msw (dive 4). Then they dived to 30 msw for 20 min with air tank. METHODS: Blood samples were collected before and after each dive. Hydrogen peroxide (H(2) O(2) ) levels, catalase (CAT) activity, mRNA expression of CAT, glutathione peroxidase (GPx) and superoxide dismutase (SOD), and the [Ca(2+) ]i in lymphocytes were measured. RESULTS: The dives slightly decreased lymphocyte number and significantly reduced lymphocyte H(2) O(2) levels. CAT activity was higher after scuba diving and, dive 3 enhanced mRNA gene expression of CAT, GPx and SOD. The [Ca(2+) ]i was higher after dive 1 and 2 than pre-diving, while was maintained at pre-diving value after dive 3 and 4. CONCLUSION: Our results suggest that pre-breathing oxygen, in particular at 12 msw, may enhance lymphocyte antioxidant activity and reduce reactive oxygen species levels. Pre-breathing oxygen in water may also preserve calcium homeostasis, suggesting a protective role in the physiological lymphocyte cell functions.

The safety of anti-TNF agents in the elderly.

Rheumatoid arthritis, ankylosing spondylitis and psoriatic arthritis are commonly thought of as inflammatory diseases that affect younger individuals. Although the initial presentation of these diseases is common in a patients twenties or thirties, they usually persist for the duration of the patients life. In addition, up to one-third of patients with RA have disease onset after 60 years of age. Anti-TNF-a therapies now have well-recognized safety profiles that have been demonstrated in the usual clinical trial populations for these diseases, but such populations under-represent patients > or =65 years of age. This retrospective study aims to determine the safety profiles for etanercept, infliximab and adalimumab in patients of 65 years or more, undergoing anti-TNF treatment for an active inflammatory disease such as rheumatoid arthritis, ankylosing spondylitis or psoriatic arthritis, or skin disease like psoriasis. Our data show that admitting elderly patients into anti-TNF therapeutic regimens is a safe option and that it grants these patients access to the best current therapeutic option, possibly leading to better disease outcome. Quality of life in elderly patients affected by arthritis or psoriasis, often reduced by comorbidities, is as important as quality of life in younger patients. Applying the recommended screening before using biological treatment helps to reduce adverse events related to the therapy, and the application of the same screening in elderly patients seems to lead to comparable results.

Extracellular guanosine and GTP promote expression of differentiation markers and induce S-phase cell-cycle arrest in human SH-SY5Y neuroblastoma cells.

SH-SY5Y neuroblastoma cells, a model for studying neuronal differentiation, are able to differentiate into either cholinergic or dopaminergic/adrenergic phenotypes depending on media conditions. Using this system, we asked whether guanosine (Guo) or guanosine-5'-triphosphate (GTP) are able to drive differentiation towards one particular phenotype. Differentiation was determined by evaluating the frequency of cells bearing neurites and assessing neurite length after exposure to different concentrations of Guo or GTP for different durations. After 6 days, 0.3 mM Guo or GTP induced a significant increase in the number of cells bearing neurites and increased neurite length. Western blot analyses confirmed that purines induced differentiation; cells exposed to purines showed increases in the levels of GAP43, MAP2, and tyrosine hydroxylase. Proliferation assays and cytofluorimetric analyses indicated a significant anti-proliferative effect of purines, and a concentration-dependent accumulation of cells in S-phase, starting after 24 h of purine exposure and extending for up to 6 days. A transcriptional profile analysis using gene arrays showed that an up-regulation of cyclin E2/cdk2 evident after 24 h was responsible for S-phase entry, and a concurrent down-regulation of cell-cycle progression-promoting cyclin B1/B2 prevented S-phase exit. In addition, patch-clamp recordings revealed that 0.3 mM Guo or GTP, after 6 day incubation, significantly decreased Na(+) currents. In conclusion, we showed Guo- and GTP-induced cell-cycle arrest in neuroblastoma cells and suggest that this makes these cells more responsive to differentiation processes that favor the dopaminergic/adrenergic phenotype.

Reduced anterior hippocampal formation volume in hyponatremic schizophrenic patients.

Diminished hippocampal volume occurs in the anterior segment of some schizophrenic patients, and in the posterior segment in others. The significance of hippocampal pathology in general and these segmental differences in specific is not known. Several lines of evidence suggest anterior hippocampal pathology underlies the life-threatening hyponatremia seen in a subgroup of patients with schizophrenia; therefore our goal was to determine if this region was preferentially diminished in hyponatremic patients. We studied seven polydipsic hyponatremic, ten polydipsic normonatremic, and nine nonpolydipsic normonatremic schizophrenic inpatients, as well as 12 healthy controls. All underwent structural scanning on a high resolution (3.0 T) magnetic resonance imaging (MRI) scanner. Hippocampal formation, amygdala, and third ventricle volumes were manually traced in each subject. The hippocampus was divided at the posterior extent of the uncus, and all structural volumes were corrected for whole brain volume and other significant recognized factors (i.e., age, gender, height, parental education). Despite being overhydrated, anterior hippocampal formation volume was diminished in those with polydipsia and hyponatremia relative to each of the other three groups. Third ventricle volume was larger in this group than in healthy controls but similar to the two patient groups. Posterior hippocampal and amygdala volumes did not differ between groups. Other potential confounds (e.g., water imbalance) either had no effect or accentuated these differences. We conclude the anterior hippocampal formation is smaller in hyponatremic schizophrenic patients, thereby linking an important and objective clinical feature of schizophrenia to a neural pathway that can be investigated in animal models. The findings strengthen the hypothesis that anterior hippocampal formation pathology disrupts functional connectivity with other limbic structures in schizophrenia.

Bullying behaviors among US youth: prevalence and association with psychosocial adjustment.

CONTEXT: Although violence among US youth is a current major concern, bullying is infrequently addressed and no national data on the prevalence of bullying are available. OBJECTIVES: To measure the prevalence of bullying behaviors among US youth and to determine the association of bullying and being bullied with indicators of psychosocial adjustment, including problem behavior, school adjustment, social/emotional adjustment, and parenting. DESIGN, SETTING, AND PARTICIPANTS: Analysis of data from a representative sample of 15 686 students in grades 6 through 10 in public and private schools throughout the United States who completed the World Health Organization's Health Behaviour in School-aged Children survey during the spring of 1998. MAIN OUTCOME MEASURE: Self-report of involvement in bullying and being bullied by others. RESULTS: A total of 29.9% of the sample reported moderate or frequent involvement in bullying, as a bully (13.0%), one who was bullied (10.6%), or both (6.3%). Males were more likely than females to be both perpetrators and targets of bullying. The frequency of bullying was higher among 6th- through 8th-grade students than among 9th- and 10th-grade students. Perpetrating and experiencing bullying were associated with poorer psychosocial adjustment (P<.001); however, different patterns of association occurred among bullies, those bullied, and those who both bullied others and were bullied themselves. CONCLUSIONS: The prevalence of bullying among US youth is substantial. Given the concurrent behavioral and emotional difficulties associated with bullying, as well as the potential long-term negative outcomes for these youth, the issue of bullying merits serious attention, both for future research and preventive intervention.

Aitken-based acceleration methods for assessing convergence of multilayer neural networks.

This paper first develops the ideas of Aitken delta(2) method to accelerate the rate of convergence of an error sequence (value of the objective function at each step) obtained by training a neural network with a sigmoidal activation function via the backpropagation algorithm. The Aitken method is exact when the error sequence is exactly geometric. However, theoretical and empirical evidence suggests that the best possible rate of convergence obtainable for such an error sequence is log-geometric. This paper develops a new invariant extended-Aitken acceleration method for accelerating log-geometric sequences. The resulting accelerated sequence enables one to predict the final value of the error function. These predictions can in turn be used to assess the distance between the current and final solution and thereby provides a stopping criterion for a desired accuracy. Each of the techniques described is applicable to a wide range of problems. The invariant extended-Aitken acceleration approach shows improved acceleration as well as outstanding prediction of the final error in the practical problems considered.