N-acetyl-L-cysteine improves the performance of chronic cyclic heat-stressed finisher broilers but has no effect on tissue glutathione levels.

1. It was hypothesised that dietary N-acetyl-L-cysteine (NAC) in feed, as a source of cysteine, could improve the performance of heat-stressed finisher broilers by fostering glutathione (GSH) synthesis. GSH is the most abundant intracellular antioxidant for which the sulphur amino acid cysteine is rate limiting for its synthesis.2. In the first experiment, four levels of NAC: 0, 500, 1000 and 2000 mg/kg were added to a diet with a suboptimal level of sulphur amino acids in the finisher phase. In the second experiment, NAC was compared to other sulphur amino acid sources at equal molar amounts of digestible sulphur amino acids. Birds were allocated to four groups: control, 2000 mg/kg NAC, 1479 mg/kg L-cystine, and 2168 mg/kg Ca-salt of 2-hydroxy-4-(methylthio)butanoic acid. A chronic cyclic heat stress model (temperature was increased to 34°C for 7 h daily) was initiated at 28 d of age.3. In the first experiment, growth performance and feed efficiency in the finisher phase were significantly improved by graded NAC. ADG was 88.9, 92.2, 93.7 and 97.7 g/d, and the feed-to-gain ratio was 2.18, 1.91, 1.85 and 1.81 for the 0, 500, 1000 and 2000 mg/kg NAC treatments, respectively. However, liver and heart GSH levels were not affected by NAC. On d 29, liver gene transcript of cystathionine-beta-synthase like was reduced by NAC, which suggested reduced trans-sulphuration activity. The second experiment showed that L-cystine and Ca-salt of 2-hydroxy-4-(methylthio) butanoic acid were more effective in improving performance than NAC.4. In conclusion, N-acetyl-L-cysteine improved dose-dependently growth and feed efficiency in heat-stressed finishing broilers. However, this was not associated with changes in tissue GSH levels, but more likely worked by sparing methionine and/or NAC's and cysteine's direct antioxidant properties.

Effects of methionine and guanidinoacetic acid supplementation on performance and energy metabolites in breast muscle of male broiler chickens fed corn-soybean diets.

1. Guanidinoacetic acid (GAA) is the single endogenous precursor of creatine, which plays a critical role in energy homeostasis of cells. Since GAA is endogenously converted to creatine by methylation, it was hypothesised that the effects of dietary GAA supplementation might determine the methionine (Met) availability in corn-soybean based diets. 2. A total of 540, one-day-old male Ross 308 broilers were allocated to nine dietary treatments with six replicates (10 birds each) in a 3 × 3 factorial arrangement with three graded levels of supplementary Met (+0.4 g/kg per level), whilst cystine was equal across groups, resulting in a low, medium and high level of total sulphur amino acids, and with three levels of GAA (0, 0.6 and 1.2 g/kg). Birds were fed for 42 days. 3. Increasing levels of supplemental Met enhanced performance indices in all rearing periods, although there was no effect on feed conversion ratio in the grower or feed intake in the finisher periods. Final body weight was 8.8% and 14.6% higher in the birds fed medium and high Met diets, respectively, compared to the low Met level. Relative breast weight and protein content in muscle on d 25 linearly increased with higher levels of Met. At low and high Met levels, growth in the finisher phase was negatively affected by supplementing GAA at 1.2 g/kg. It was suggested that disturbances in methylation homeostasis and/or changes in Arg metabolism might explain these findings. At the end of the grower phase, muscle creatine content was higher when feeding GAA at 0.6 and 1.2 g/kg (4464 and 4472, respectively, vs. 4054 mg/kg fresh muscle in the control group). 4. The effects of dietary GAA supplementation were influenced by the dietary Met level only in the finisher period, which indicates the need for proper sulphur amino acid formulation in diets when feeding GAA.

Guanidinoacetic acid supplementation in broiler chickens fed on corn-soybean diets affects performance in the finisher period and energy metabolites in breast muscle independent of diet nutrient density.

(1) Guanidinoacetic acid (GAA) is the single immediate endogenous precursor of creatine (Cr). It was hypothesised that dietary GAA would have different effects on performance and energy metabolites in breast muscle depending on the nutrient density (ND) of corn-soybean-based diets. (2) A total of 540 one-day-old male Ross 308 broilers were allocated to 9 dietary treatments with 6 replicates (10 birds each) in a 3 × 3 factorial arrangement with three levels of ND (low, 2800; medium, 2950 and high, 3100 kcal metabolizable energy (ME)/kg; and with the other nutrients being constant relative to ME) and supplemented with three levels of GAA (0, 0.6 and 1.2 g/kg) in a 42-d feeding trial. (3) In the starter and grower periods, increasing levels of ND improved body weight (BW), average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR), with the exception of ADFI in the starter period. GAA supplementation did not affect performance characteristics. All performance indicators responded markedly to increasing ND in the finisher period, whereas the highest GAA level reduced ADFI compared to the unsupplemented control (156 vs. 162 g/d) and concomitantly FCR (1.81 vs. 1.93). No interactive effects were noted for any performance trait. The high ND diet resulted in more breast meat yield on d42, associated with higher fat content and darker colour compared to the other ND levels. The GAA supplementation did not affect carcass and breast traits. At the end of the experiment, Cr was elevated when feeding GAA at 1.2 g/kg (5455 vs. 4338 mg/kg fresh muscle). (4) To conclude, ND had a substantial effect on performance and carcass traits, whereas any effect of GAA was limited to FCR in the finisher period and independent of diet ND level.

Meningeal SWI/SNF related, matrix-associated, actin-dependent regulator of chromatin, subfamily B member 1 (SMARCB1)-deficient tumours: an emerging group of meningeal tumours.

AIMS: Bi-allelic inactivation of SWI/SNF related, matrix-associated, actin-dependent regulator of chromatin, subfamily B member 1 (SMARCB1; also known as INI1) and loss of immunohistochemical expression of SMARCB1 define the group of SMARCB1-deficient tumours. Initially highlighted in malignant rhabdoid tumours, this inactivation has subsequently been observed in several intra and extracranial tumours. To date, primary meningeal SMARCB1-deficient tumours have not been described. We report two cases of meningeal SMARCB1-deficient tumours occurring in adults. METHODS: We performed immunohistochemical analyses, comparative genomic hybridization, fluorescence in situ hybridization and targeted next-generation sequencing. RESULTS: The first meningeal tumour was a solitary mass, composed of rhabdoid, adenoid, chordoid and sarcomatoid areas. The second case presented as multiple, bilateral, supra and infratentorial nodules, was composed of fusiform and ovoid cells embedded in a myxoid stroma. Tumour cells were positive for epithelial membrane antigen (EMA), vimentin and CD34 and negative for SMARCB1 and meningothelial, melanocytic, muscular, glial markers. In the first case, one allele of SMARCB1 was completely deleted, whereas in the second case, loss of expression of SMARCB1 was observed as a consequence of a homozygous deletion of SMARCB1. CONCLUSIONS: The phenotype and genotype of these two cases did not fit diagnostically with entities already known to be SMARCB1-deficient tumours. As both tumours shared common features, they are regarded as belonging to an emerging group of primary meningeal SMARCB1-deficient tumours, not described to date. To facilitate the identification and characterization of these tumours, we recommend SMARCB1 immunohistochemistry for primary meningeal tumours which are difficult to classify, especially if immunopositive for EMA and CD34.

Controlled release of chlorhexidine from a mesoporous silica-containing macroporous titanium dental implant prevents microbial biofilm formation.

Roughened surfaces are increasingly being used for dental implant applications as the enlarged contact area improves bone cell anchorage, thereby facilitating osseointegration. However, the additional surface area also entails a higher risk for the development of biofilm associated infections, an etiologic factor for many dental ailments, including peri-implantitis. To overcome this problem, we designed a dental implant composed of a porous titanium-silica (Ti/SiO2) composite material and containing an internal reservoir that can be loaded with antimicrobial compounds. The composite material consists of a sol-gel derived mesoporous SiO2 diffusion barrier integrated in a macroporous Ti load-bearing structure obtained by powder metallurgical processing. The antimicrobial compounds can diffuse through the porous implant walls, thereby reducing microbial biofilm formation on the implant surface. A continuous release of µM concentrations of chlorhexidine through the Ti/SiO2 composite material was measured, without initial burst effect, over at least 10 days and using a 5 mM chlorhexidine solution in the implant reservoir. Metabolic staining, CFU counting and visualisation by scanning electron microscopy confirmed that Streptococcus mutans biofilm formation on the implant surface was almost completely prevented due to chlorhexidine release (preventive setup). Moreover, we demonstrated efficacy of released chlorhexidine against mature Streptococcus mutans biofilms (curative setup). In conclusion, we provide a proof of concept of the sustained release of chlorhexidine, one of the most widely used oral antiseptics, through the Ti/SiO2 material thereby preventing and eradicating biofilm formation on the surface of the dental implant. In principle, our flexible design allows for the use of any bioactive compound, as discussed.

Breast implant-associated anaplastic large cell lymphoma: two distinct clinicopathological variants with different outcomes.

BACKGROUND: ALK-negative anaplastic large cell lymphoma associated with breast implant (i-ALCL) has been recently recognized as a distinct entity. Among 43 830 lymphomas registered in the French Lymphopath network since 2010, 300 breast lymphomas comprising 25 peripheral T-cell lymphomas (PTCL) were reviewed. Among PTCL, ALK-negative ALCL was the most frequent and all of them were associated with breast implants. PATIENTS AND METHODS: Since 2010, all i-ALCL cases were collected from different institutions through Lymphopath. Immuno-morphologic features, molecular data and clinical outcome of 19 i-ALCLs have been retrospectively analyzed. RESULTS: The median age of the patients was 61 years and the median length between breast implant and i-ALCL was 9 years. Most implants were silicone-filled and textured. Implant removal was performed in 17 out of 19 patients with additional treatment based on mostly CHOP or CHOP-like chemotherapy regimens (n = 10/19) or irradiation (n = 1/19). CHOP alone or ABVD following radiation without implant removal have been given in two patients. The two clinical presentations, i.e. effusion and less frequently tumor mass correlated with distinct histopathologic features: in situ i-ALCL (anaplastic cell proliferation confined to the fibrous capsule) and infiltrative i-ALCL (pleomorphic cells massively infiltrating adjacent tissue with eosinophils and sometimes Reed-Sternberg-like cells mimicking Hodgkin lymphoma). Malignant cells were CD30-positive, showed a variable staining for EMA and were ALK negative. Most cases had a cytotoxic T-cell immunophenotype with variable T-cell antigen loss and pSTAT3 nuclear expression. T-cell receptor genes were clonally rearranged in 13 out of 13 tested cases. After 18 months of median follow-up, the 2-year overall survival for in situ and infiltrative i-ALCL was 100% and 52.5%, respectively. CONCLUSIONS: In situ i-ALCLs have an indolent clinical course and generally remain free of disease after implant removal. However, infiltrative i-ALCLs could have a more aggressive clinical course that might require additional therapy to implant removal.

Guidelines for May-Grünwald-Giemsa staining in haematology and non-gynaecological cytopathology: recommendations of the French Society of Clinical Cytology (SFCC) and of the French Association for Quality Assurance in Anatomic and Cytologic Pathology (AFAQAP).

OBJECTIVE: Since the guidelines of the International Committee for Standardisation in Haematology (ICSH) in 1984 and those of the European Committee for External Quality Assessment Programmes in Laboratory Medicine (EQALM) in 2004, no leading organisation has published technical recommendations for the preparation of air-dried cytological specimens using May-Grünwald-Giemsa (MGG) staining. DATA SOURCES: Literature data were retrieved using reference books, baseline-published studies, articles extracted from PubMed/Medline and Google Scholar, and online-available industry datasheets. RATIONALE: The present review addresses all pre-analytical issues concerning the use of Romanowsky's stains (including MGG) in haematology and non-gynaecological cytopathology. It aims at serving as actualised, best practice recommendations for the proper handling of air-dried cytological specimens. It, therefore, appears complementary to the staining criteria of the non-gynaecological diagnostic cytology handbook edited by the United Kingdom National External Quality Assessment Service (UK-NEQAS) in February 2015.

[Creation and normalisation of a verbal episodic memory task in elderly adults: "GERIA-12"]. / Élaboration et normalisation d'une épreuve d'évaluation de la mémoire épisodique verbale chez la personne âgée: "GERIA-12".

INTRODUCTION: Early damage to episodic memory encoding and consolidation processes has been demonstrated in dementia of the Alzheimer type. However, in the domain of verbal episodic memory assessment, there are few diagnostic tools adapted to the old and oldest old as far as ease of administration and accuracy of normative data are concerned. Classic tasks are either too effortful (like the free recall/cued recall of 16 items), not sensitive enough (like the 5 words test), or insufficiently accurate for people above 70 years old in terms of normative data. AIM: The aim of this study was to develop a reduced task (in terms of number of items and number of trials) assessing verbal episodic memory in people aged between 70 and 89 years old. METHODS: The task (GERIA-12) used the same procedure as the RL/RI-16 task but the list comprised only 12 words and there were only 2 learning trials. In order to assess consolidation processes, we included 2 delayed recall trials, one after 20 minutes and the other after 24 hours. We also calculated indexes adapted from the Item-Specific Deficit Approach developed by Wright et al., which has the advantage of providing measures specific to encoding, consolidation and retrieval processes. Standardization was done with data from 220 people aged between 70 and 89 years old and belonging to 3 education levels. RESULTS: We obtained a significant effect of age and education level: scores decrease with age and increase with education. Norms have thus been calculated taking those two variables into consideration. Concerning the standardization, Barona method has been used for free recall scores while percentiles have been used for all other scores (total recall, free recall, encoding, consolidation and retrieval indexes). Normative data are also provided for intrusions and perseverations. CONCLUSION: This new task allows encoding, consolidation and retrieval processes assessment in older people and has the following advantages: the procedure is more suitable (ease and time of administration), there are accurate normative data for old and oldest old people, and there are normative data for two delayed recalls (at 20 minutes and at 24 hours).

Antioxidant enzyme activities, plasma hormone levels and serum metabolites of finishing broiler chickens reared under high ambient temperature and fed lemon and orange peel extracts and Curcuma xanthorrhiza essential oil.

The negative effects of high ambient temperature during some months of the year on poultry production have been of great concern in many countries. Dietary modifications are among the most practical ways to alleviate the effects of high temperature. Possible effects of dietary supplementation with 200 or 400 mg/kg feed of lemon peel extract (LPE), orange peel extract (OPE) and Curcuma xanthorrhiza essential oil (CXEO) under hot conditions (34 °C with 50% relative humidity for 5 h daily starting from day 28 until day 38 of age) on blood antioxidant enzyme activities, biochemical parameters and antibody titres of broiler chickens were investigated. All extracts are rich in phenolic compounds and highly available. Compared to control, supplementation with OPE at 400 mg/kg and CXEO significantly increased erythrocyte glutathione peroxidase and superoxide dismutase activity, plasma growth hormone concentrations and serum phosphorus, total protein and chloride concentrations and decreased serum low-density lipoprotein and cholesterol concentrations in chickens at 38 days of age. Regarding antibody titres, CXEO supplementation at 400 mg/kg caused a significant increase in bronchitis antibody titres. Supplementation with LPE and OPE gave more inconsistent results. Most interesting, 400 mg/kg LPE significantly increased 3,5,3'-triiodothyronine and GH concentration as compared to the control. In conclusion, the herbal extracts tested in this study, in particular CXEO at 400 mg/kg, may relieve some of the changes in blood composition induced by increased ambient temperatures.

Gene expression of heat shock protein 70 and antioxidant enzymes, oxidative status, and meat oxidative stability of cyclically heat-challenged finishing broilers fed Origanum compactum and Curcuma xanthorrhiza essential oils.

Heat stress in poultry is a serious problem in many countries and has been associated with oxidative stress. Hence, nutritional interventions with antioxidants might be beneficial. Therefore, the effects of dietary Curcuma xanthorrhiza (CX) and Origanum compactum (OC) essential oils on mRNA levels of heat shock protein 70 and antioxidant enzymes, oxidative status, and meat oxidative stability of heat-challenged broilers were studied. Starting on d 25 of age, a control diet and 4 diets containing 200 or 400 mg/kg feed of CX or OC (CX200, CX400, OC200, OC400 diets) were fed to 3 pen replicates of 20 Ross 308 chickens each. From d 28 of age on, the temperature was increased from 22 to 34°C with 50% RH for 5 h daily during 2 wk. Dietary CX or OC did not affect zootechnical performance. Feeding CX400 and both levels of OC increased the a* value in stored breast meat (P < 0.05), and OC diets tended to decrease the thiobarbituric acid reactive substances values in fresh breast meat (P = 0.061). Compared with control, at d 31, feeding CX400 and OC400 reduced mRNA levels of heat shock protein 70 and increased mRNA levels of catalase in kidney and liver (P < 0.05). The mRNA levels of superoxide dismutase were increased at d 31 on the OC400 diet in kidney and on the CX400 diet in heart (P < 0.05). In heart, at d 31, both dietary levels of CX and OC200 resulted in higher glutathione peroxidase activity (P < 0.05). Feeding CX400 increased superoxide dismutase activity in liver, kidney, and heart at d 31 (P < 0.05). Catalase activity was increased in the CX200 and OC400 groups at d 42 (P < 0.05). Feeding CX at both levels and OC200 decreased plasma malondialdehyde concentrations at d 42 (P < 0.05). In conclusion, dietary essential oils rich in simple phenolic compounds offer potential for improving the antioxidant defense against heat stress-induced changes.

Nutritional interventions to prevent and rear low-birthweight piglets.

Selection for hyperprolific sows, as a means of increasing litter size and profit, has resulted in an increased number of low-birthweight (LBW) piglets. These LBW piglets might suffer from increased morbidity and mortality during the early neonatal period. In addition, they show reduced growth performance, meat and carcass quality, which leads to an important economic loss for the farmer in the post-natal period. Therefore, nutritional interventions can be undertaken to prevent and rear LBW piglets. In the first part of this review, the preventive strategies at the sow level will be discussed. Approaches in preventing LBW piglets are to optimize the intrauterine environment via supplementing the sow during gestation. In the second part of this review, the interventions at the piglet level will be described. To increase the survival and growth rates of LBW piglets, one must focus on ensuring adequate colostrum and milk intake. Interventions include supplementing piglets, split nursing, split weaning and cross-fostering. Additional interventions increasing the probability of optimal post-natal food intake will be discussed.

CXCR7-mediated progression of osteosarcoma in the lungs.

BACKGROUND: Osteosarcoma (OS) is the most frequent primary malignant bone tumour in children and adolescents with a high propensity for lung metastasis. Chemokines and chemokine receptors have been described to have an important role in many malignancies including OS. The aim of this study was to investigate the expression of CXCR7 receptor in OS tissues and its role in the progression of the disease in the lungs. METHODS: Immunohistochemistry was used to study CXCR7 expression in primary tumours and metastatic tissues from patients with OS. Its contribution to tumour expansion in the lungs has been also assessed using animal models and synthetic-specific CXCR7 ligands. RESULTS: CXCR7 was expressed on human primary bone tumours and on lung metastases. Its expression was predominantly located on tumour-associated blood vessels. Mice challenged with OS cells and systematically treated with synthetic CXCR7 ligands presented a significant reduction of lung nodules compared with untreated mice. CONCLUSION: This study shows that CXCR7 has a critical role in OS progression in the lungs, where are expressed CXCR7 ligands, especially CXCL12. Moreover, we highlight that synthetic CXCR7 ligands could represent a powerful therapeutic tool to impede lung OS progression.

Effect of fermented moist feed on performance, gut bacteria and gut histo-morphology in broilers.

1. Fermented feed has been shown to be beneficial in pig nutrition as a tool to reduce gut microbial disorders. Experiments with fermented feed in poultry are scarce, probably because of the belief that wet feed is less suitable for this species and causes wet litter. 2. A total of 280 one-d-old Ross 308 chickens were used in a completely randomised design with two dietary treatments (7 replicates of 20 birds/treatment); air-dry feed versus the same feed in moist form (water:feed ratio of 1.3:1, on a weight basis), inoculated with Lactobacillus plantarum NCIMB 40087 (9 log10 CFU/kg feed) and batch-fermented for 48 h at 26°C. The birds were given starter (d 0-13), grower (d 4-26) and finisher (d 27-39) diets ad libitum. At the end of the grower and finisher period, two birds per pen were removed to sample intestinal contents for cultivating bacteria and intestinal tissue to determine villus height and crypt depth. 3. Fermented moist feed (FMF) batches showed good characteristics, with a pH between 3.9 and 4.4 and DL-lactic acid between 137 and 286 mmol/l. Daily feed intake and gain were reduced considerably in the FMF group in the starter (-40 and -44%, respectively) and grower (-23 and -16%) period, though in the finisher period these birds performed better, with an improved feed utilisation. Concomitant with the latter, villus height at the mid-jejunum and mid-ileum on d 39 was higher (+22.6% and +16.0%). Significantly more Lactobacilli and less coliforms were found in the foregut and less Streptococci in ileum and caeca of birds given FMF. 4. This trial showed that FMF was detrimental for early bird growth but affected beneficially feed efficiency, the composition of the gut bacteria and villus height in the small intestine in the finisher period in broilers.

Effects of feeding guanidinoacetic acid on oxidative status and creatine metabolism in broilers subjected to chronic cyclic heat stress in the finisher phase.

Dietary guanidinoacetic acid (GAA) has been shown to affect creatine (Cr) metabolic pathways resulting in increased cellular Cr and hitherto broiler performances. Yet, the impact of dietary GAA on improving markers of oxidative status remains equivocal. A model of chronic cyclic heat stress, known to inflict oxidative stress, was employed to test the hypothesis that GAA could modify bird's oxidative status. A total of 720-day-old male Ross 308 broilers were allocated to 3 treatments: 0, 0.6 or 1.2 g/kg GAA was added to corn-SBM diets and fed for 39 d, with 12 replicates (20 birds each) per treatment. The chronic cyclic heat stress model (34°C with 50-60% RH for 7 h daily) was applied in the finisher phase (d 25-39). Samples from 1 bird per pen were taken on d 26 (acute heat stress) and d 39 (chronic heat stress). GAA and Cr in plasma were linearly increased by feeding GAA on either sampling day, illustrating efficient absorption and methylation, respectively. Energy metabolism in breast and heart muscle was greatly supported as visible by increased Cr and phosphocreatine: ATP, thus providing higher capacity for rapid ATP generation in cells. Glycogen stores in breast muscle were linearly elevated by incremental GAA, on d 26 only. More Cr seems to be directed to heart muscle as opposed to skeletal muscle during chronic heat stress as tissue Cr was higher in heart but lower in breast muscle on d 39 as opposed to d 26. The lipid peroxidation marker malondialdehyde, and the antioxidant enzymes superoxide dismutase and glutathione peroxidase showed no alterations by dietary GAA in plasma. Opposite to that, superoxide dismutase activity in breast muscle was linearly lowered when feeding GAA (trend on d 26, effect on d 39). Significant correlations between the assessed parameters and GAA inclusion were identified on d 26 and d 39 using principal component analysis. To conclude, beneficial performance in heat-stressed broilers by GAA is associated with enhanced muscle energy metabolism which indirectly may also support tolerance against oxidative stress.

CXCR7 receptors facilitate the progression of colon carcinoma within lung not within liver.

BACKGROUND: Liver and lung metastases are the predominant cause of colorectal cancer (CRC)-related mortality. Chemokine-receptor pairs have a critical role in determining the metastatic progression of tumours. Our hypothesis was that disruption of CXCR7/CXCR7 ligands axis could lead to a decrease in CRC metastases. METHODS: Primary tumours and metastatic tissues from patients with CRC were tested for the expression of CXCR7 and its ligands. Relevance of CXCR7/CXCR7 ligands for CRC metastasis was then investigated in mice using small pharmacological CXCR7 antagonists and CRC cell lines of human and murine origins, which - injected into mice - enable the development of lung and liver metastases. RESULTS: Following injection of CRC cells, mice treated daily with CXCR7 antagonists exhibited a significant reduction in lung metastases. However, CXCR7 antagonists failed to reduce the extent of liver metastasis. Moreover, there were subtle differences in the expression of CXCR7 and its ligands between lung and liver metastases. CONCLUSION: Our study suggests that the activation of CXCR7 on tumour blood vessels by its ligands may facilitate the progression of CRC within lung but not within liver. Moreover, we provide evidence that targeting the CXCR7 axis may be beneficial to limit metastasis from colon cancer within the lungs.

Utility of Ki67 immunostaining in the grading of pineal parenchymal tumours: a multicentre study.

AIMS: Pineal parenchymal tumours (PPTs) are rare neoplasms that are divided into pineocytoma (PC), pineoblastoma (PB) and PPT of intermediate differentiation (PPTID). Factors affecting the survival of patients with PPTs are morphological subtype and histological grading according to mitotic index and neurofilament immunostaining. Grading criteria to distinguish PPTIDs are difficult to define, particularly when using small specimens. The Ki67 labelling index (LI) might be helpful in distinguishing between grade II and III PPTIDs. Our study was performed to assess the predictive value of the Ki67 LI in a large cooperative series of PPTs and to evaluate whether inclusion of this data would improve and refine the World Health Organization classification. METHODS: A retrospective analysis of 33 PPTs was performed. The histological features of the tumours were reviewed and Ki67 LI scoring was evaluated by immunohistochemistry. Data were correlated with the patients' survival. RESULTS: The mean Ki67 LI was significantly different for tumour grades (0 in PC, 5.2 ± 0.4 in PPTID grade II, 11.2 ± 2.0 in PPTID grade III, 36.4 ± 6.2 in PB; P < 0.0001). However, there was no statistically significant difference in either overall or disease-free survival evaluated by the Kaplan-Meier method for patients with different grade tumours or Ki67 LI, possibly due to the different clinical management of patients in different centres. CONCLUSIONS: The Ki67 LI may be a useful additional tool for grading PPTs, more particularly in small tumour samples.

Supplementation of guanidinoacetic acid to broiler diets: effects on performance, carcass characteristics, meat quality, and energy metabolism.

Creatine, (CREA) a central constituent in energy metabolism, is obtained from dietary animal protein or de novo synthesis from guanidinoacetic acid (GAA). Especially in all-vegetable diets, supplemental CREA or GAA may restore the CREA availability in tissues, and hence, improve performance. In this study, 768 one-d-old male Ross 308 broilers were assigned to 1 of 4 diets: negative control, all-vegetable corn-soybean-based; negative control supplemented with either 0.6 or 1.2 g of GAA per kilogram of feed; and positive control (60, 30, and 30 g/kg of fish meal in the starter, grower, and finisher diets, respectively). Each treatment was replicated in 6 pens of 32 birds each. At the end of the grower period (d 26), 2 birds per pen were euthanized for metabolic measurements. Four broilers per pen were selected at slaughter age (d 39) to determine carcass characteristics and meat quality. Compared with the negative control, GAA supplementation resulted in an improved gain:feed ratio (P < 0.05) and ADG (P < 0.05; + 2.7 and + 2.2% for GAA at 0.6 and 1.2 g/kg, respectively) throughout the entire period. Breast meat yield was higher for the GAA diets compared with that of the negative control birds (P < 0.05; 30.6 vs. 29.4%) and was comparable with that of the positive control birds (30.2%). With regard to meat quality, lower ultimate pH values, higher cooking and press fluid losses, and higher color L* values were observed for the GAA diets compared with those of the negative control diet (P < 0.05). These effects were small, however. The GAA and CREA levels in breast meat were lower and higher, respectively, in GAA-fed birds compared with those of the control birds (P < 0.01). The diets did not affect plasma metabolic traits, except that plasma insulin-like growth factor I concentrations were almost twice as high in animals fed 1.2 g/kg of GAA compared with those of all other treatments. The GAA included in all-vegetable diets improved animal performance for the whole rearing period and increased breast meat yield.

Intact brown seaweed (Ascophyllum nodosum) in diets of weaned piglets: effects on performance, gut bacteria and morphology and plasma oxidative status.

The aim was to assess the effects of intact dried Ascophyllum nodosum seaweed on piglet performances, gut bacteria and function and plasma oxidative status. A total of 160 weaned piglets (21 days, 6.59 ± 0.91 kg) were allocated to four dietary treatments with eight pen replicates of five animals each for 28 days: a control diet; based on cereals, soybean meal and milk products, and three basal diets supplemented with either 2.5, 5.0 or 10.0 g dried seaweed per kg. At day 12/13 one piglet from each pen was sacrificed. Plasma samples were taken to determine parameters of oxidative status. Digesta were sampled for microbiological plate countings onto selective media and molecular analysis using PCR-DGGE. Small intestinal tissue was taken for morphological and electro-physiological determinations. Data were analysed by a linear model with treatment as fixed effect. A. nodosum supplementation had no effect on daily weight gain, nor did it alter feed conversion ratio. Plate countings failed to reveal differences among treatments. Dendograms prepared using PCR-DGGE banding patterns did not indicate clustering of microbial profiles based on diet supplement. Plasma oxidative status and outcome of morphology and of electro-physiological measurements from gut tissues were similar for all treatments. Thus, the addition of A. nodosum seaweed to well digestible diets did not enhance performances of piglets nor some gut health parameters and plasma oxidative status.

Ecology and evolution of antibiotic persistence.

Bacteria have at their disposal a battery of strategies to withstand antibiotic stress. Among these, resistance is a well-known mechanism, yet bacteria can also survive antibiotic attack by adopting a tolerant phenotype. In the case of persistence, only a small fraction within an isogenic population switches to this antibiotic-tolerant state. Persistence depends on the ecological niche and the genetic background of the strains involved. Furthermore, it has been shown to be under direct and indirect evolutionary pressure. Persister cells play a role in chronic infections and the development of resistance, and therefore a better understanding of this phenotype could contribute to the development of effective antibacterial therapies. In the current review, we discuss how ecological and evolutionary forces shape persistence.

In vitro stability and ex vivo absorption of thymol monoglucosides in the porcine gut.

Thymol α-D-glucopyranoside (TαG) and thymol ß-D-glucopyranoside (TßG) are believed to have different kinetic behaviours in the porcine gut than its parent aglycon thymol. However, recently, it was shown that concentrations of both glucosides decreased rapidly in the stomach and proximal small intestine following oral supplementation to piglets as did thymol. Yet, the stability of thymol glucosides in gut contents and their absorption route remains obscure. Therefore, a series of in vitro incubations were performed, simulating the impact of pH, digestive enzymes, bacterial activity and mucosal extracts on stability of these glucosides. Their absorption mechanisms were investigated using the Ussing chamber model in the presence or the absence of inhibitors of sodium-dependent glucose linked transporter 1 and lactase phlorizin hydrolase. Both glucosides remained intact at physiological pH levels in the presence of digestive enzymes. Recoveries from TαG and TßG were below 90% when incubated with small intestinal homogenates from the distal jejunum or from all sampled sites, respectively. However, no aglycon could be detected in these samples. Bacterial inoculum of the small intestine, on the other hand, hydrolysed TßG quickly with up to 44% of free aglycon appearing. TαG proved more resistant to porcine gastro-intestinal bacterial glucosidases with only trace amounts (<1%) of free thymol at the end of the incubations. Electrophysiological measurements in Ussing chambers did not suggest active transport of the glucosides. Mucosal TαG and TßG concentrations were unchanged between start and end of the absorption measurements. Additionally, no TαG and only a very limited amount of TßG were retrieved from the serosal side. Tissue associated concentrations, although marginal (<1% of luminal concentration), were mainly as intact glucoside or as aglycon for TαG and TßG, respectively. Addition of both inhibitors significantly increased the amount of intact glucosides retrieved from the mucosal tissues as compared to controls. In conclusion, bacterial hydrolysis was identified as the most important source of TßG loss, whereas TαG seemed less prone to degradation or absorption in these in vitro and ex vivo models.