Total replacement of soybean meal with alternative plant-based ingredients and a combination of feed additives in broiler diets from 1 day of age during the whole growing period.

The capacity of combinations of feed enzymes, natural betaine and a probiotic, combined with alternative plant-based ingredients, to totally replace soybean meal (SBM) in a broiler diet was evaluated. Day-old Ross 308 males (2,574) were assigned to 9 treatments (13 pens/treatment, 22 birds/pen) in a completely randomized design. All diets were pelleted and fed ad libitum in 4 phases: starter, grower, finisher 1, finisher 2 (0-10, 10-21, 21-35, and 35-42 d of age, respectively). Treatments included: 1) control diet containing SBM (SBM control), supplemented with phytase (PhyG), at 2,000, 1,500, 1000 and 1,000 FTU/kg in each phase and xylanase (X) at 750 U/kg, [crude protein (CP): 23.5%, 22.0%, 20.2% and 19.3% in each phase]; 2) to 5), alternative (ALT), SBM-free diets, containing the same CP level as the control ("CP high"), supplemented with PhyG as in the control, protease (P, 800 U/kg) and in 2) xylanase (750 U/kg) (ALT+PhyG+P+X), 3) xylanase-ß-glucanase (XB, 1,200 U/kg and 152 U/kg) (Alt+PhyG+P+XB), 4) XB plus betaine (800 g/ton) (ALT+PhyG+P+XB+Bet), and 5) XB plus a probiotic [150,000 colony forming units (CFU)/g] (ALT+PhyG+P+XB+Prob); 6) to 9) as treatments 2) to 5) but with CP reduced by -2.0 to -1.5% points vs. control ('CP low'). Final (d 42) BW and overall (d 0-42) feed conversion ratio (FCR) of birds fed the SBM control exceeded breeder objectives (+3.8% and -1.9%, respectively). Overall FCR was reduced and d 42 BW increased in birds fed "low" vs. "high" CP (P < 0.01). Overall FCR and feed intake were not different in ALT+PhyG+XB+P+Bet and ALT+PhyG+XB+P+Prob vs. the control, whereas final BW was reduced (P < 0.05) in all ALT treatments but close to breeder objectives (98.3%) in ALT+PhyG+XB+P+Prob. Feed costs of this treatment were similar to the control. Total replacement of SBM with alternative plant-based ingredients in a CP-low diet supplemented with hydrolytic enzymes and probiotics can achieve growth performance outcomes close to commercial breeder objectives.

Modeling improvements in ileal digestible amino acids by a novel consensus bacterial 6-phytase variant in broilers.

Data from 13 datasets from 4 trials on the effect of a novel consensus bacterial 6-phytase variant (PhyG) on the apparent ileal digestibility (AID) of amino acids (AA) in broilers were used to model AID AA responses. The datasets were obtained from 3 trial locations (New Zealand, Australia and United States) and collectively incorporated variations in diet composition (feedstuff composition, phytate-P (PP) level, limestone solubility), feed form (mash or pellet), bird genetics (strain), and age at sampling (11-35 d of age). In total, 384 observations were analyzed. First, the relationships between AID of AA (as coefficients) and increasing phytase dose level from 0 to 4,000 FTU/kg were evaluated across all datasets using exponential curve fitting. Second, the percentage unit change in AID of AA at each phytase dose level from baseline (basal diet [BD] without phytase) was calculated separately for each dataset and the data then modeled together using exponential curve fitting. The model-predicted mean coefficient of AID of total AA in basal diets was 0.76 (range 0.56 [Cys] to 0.83 [Glu]), which was increased by PhyG to 0.80 and 0.81 at 2,000 and 4,000 FTU/kg, respectively. Exponential increases in the percentage unit improvement in AID of 18 individual and of total AA with increasing phytase dose level were evident (P < 0.05). Improvements (vs. BD) at 2,000 FTU/kg and 4,000 FTU/kg, respectively, were greatest for Cys (+9.2 and +11.0% units), Met (after deduction of synthetic Met, +8.4 and +9.0% units), and Thr (after deduction of synthetic Thr, +6.2 and +7.3% units). The data demonstrated consistent improvements in the AID of AA by the phytase. The modeling results generated from data gathered from birds sampled at different ages and from different dietary settings with correction of synthetic AA for Lys, Met, Thr, and Trp, enabled a more accurate prediction of the digestible AA contribution from the diet by this novel phytase. This will allow diet-specific AA matrix recommendations to be made in commercial feed formulations.

Effect of increasing dose level of a novel consensus bacterial 6-phytase variant on phytate degradation in broilers fed diets containing varied phytate levels.

1. The effect of increasing the dose level of a novel consensus bacterial 6-phytase variant on apparent ileal digestibility (AID) of phosphorus (P), phytic acid (inositol hexa-phosphate, IP6) and ileal IP6 degradation profile was studied in diets containing varying phytate-P (PP) levels.2. Ross 308, one-day-old males (n = 1,800) were allocated to cages (20 birds/cage, six cages/treatment) in a completely randomised design employing a 3 × 5 factorial arrangement (three PP levels: 2.45 (low) 2.95 (medium) and 3.45 g/kg (high); five dose levels of phytase (PhyG): 0, 500, 1,000, 2,000 and 4,000 FTU/kg). Phased diets were based on wheat, corn, soybean meal, rapeseed meal and rice bran (d 0 to 10; 2.60 g/kg digestible P, 7.6 g/kg calcium (Ca); d 11 to 21; 2.10 g/kg digestible P, 6.4 g/kg Ca). Ileal digesta was collected on d 21 for determination of P, IP6 and IP-esters content. Data were analysed by factorial ANOVA; means separation was achieved using Tukey's HSD test.3. Increasing PP reduced AID of IP6 and sum of IP3-6 (%) (P < 0.05) but absolute P-release (g/kg diet) above NC was increased (P < 0.05) at high vs. low PP. Increasing phytase dose exponentially increased (P < 0.001) AID IP6, sum of IP3-6 (%) and digestible IP3-6-P g/kg diet (P < 0.001). AID P was increased but there was an interaction with PP level (P < 0.001). Ileal accumulation of IP5-3-P was universally low with PhyG at ≥1,000 FTU/kg (<0.06 g/100 g DM). At 2,000 and 4,000 FTU/kg, AID IP6 was 97.2, 92.7, 92.6% and 100, 97.2, 97.1%, respectively, at low, medium and high PP. At 2,000 FTU/kg, phytate-P release estimated as the increase (above NC) in ileal digestible sum of IP3-6-P in the diet was 2.26, 2.59 and 3.10 g/kg in low, medium and high PP, respectively.4. The data demonstrated that the novel phytase was effective in breaking down phytate to low IP-esters in diets with varied PP content but the optimal dose level for maximising P-release may differ in diets with varying PP content.

Thromboembolic events and thromboprophylaxis associated with immunomodulators in multiple myeloma patients: a real-life study.

PURPOSE: The aim of this study is to assess international guidelines implementation concerning thromboprophylaxis strategy in myeloma patients treated with immunomodulatory drugs. METHODS: This retrospective study includes multiple myeloma patients treated with immunomodulatory drugs between 2014 and 2017 in the Hematology department of a teaching hospital (Hospices Civils de Lyon, France) and followed by the multidisciplinary care plan for cancer outpatients ONCORAL (ONCological care for outpatients with ORAL anticancer drugs). Data from immunomodulatory drugs administration, thromboprophylaxis strategy and thrombotic events were collected from medical files. Adherence to 2010 International Myeloma Working Group (IMWG) guidelines was assessed. RESULTS: 213 patients received at least one immunomodulatory drug: lenalidomide (60.9%), pomalidomide (24.0%) and thalidomide (15.1%). About two third of treatment lines (66.2%) were in accordance with IMWG recommendations. Among the others, 30.5% and 69.5% had thromboprophylaxis, respectively, superior or inferior to IMWG recommendations. 37 venous thrombotic events and 4 arterial thromboembolisms (one patient experienced both a stroke and deep venous thrombosis simultaneously) were reported. CONCLUSION: Thromboprophylaxis was systematically performed in myeloma patients treated with immunomodulatory drugs in this real-life retrospective cohort. However, the choice of anticoagulant or anti-platelet agent remains debatable, as adherence to existing guidelines was variable.

Nerve-specific extracellular matrix hydrogel promotes functional regeneration following nerve gap injury.

Nerve transection requires surgical intervention to restore function. The standard of care involves coaptation when a tension-free repair is achievable, or interposition of a graft or conduit when a gap remains. Despite advances, nerve gap injury is associated with unsatisfactory recovery. This study investigates the use of a decellularized, porcine nerve-derived hydrogel filler (peripheral nerve matrix, PNM) for conduits in an 8 mm rat sciatic nerve gap model. The decellularized tissue maintained multiple nerve-specific matrix components and nerve growth factors. This decellularized tissue was used to formulate hydrogels, which were deployed into conduits for nerve gap repair. Nerve recovery was assessed up to 24 weeks post injury by gait analysis, electrophysiology, and axon counting. Deployment of PNM within conduits was shown to improve electrophysiologic response and axon counts compared with those of empty conduit controls. These results indicate that PNM has potential benefits when used as a filler for conduits in nerve gap injuries.

Effect of a novel consensus bacterial 6-phytase variant in grower pigs fed corn-soybean meal-based diets formulated with a full nutrient matrix and no added inorganic phosphorus.

The capacity of a novel consensus bacterial 6-phytase variant (PhyG) to entirely replace dietary inorganic phosphorus (Pi) source in grower pigs fed diets with reduction of calcium (Ca), net energy (NE), and digestible amino acids (AA) was evaluated, using growth performance and apparent total tract digestibility (ATTD) of nutrients as outcome measures. A total of 352 mixed-sex pigs (initial BW 23.4 kg) were randomized to 4 treatments, 8 pigs/pen, and 11 pens/treatment. Diets were corn-soybean meal-based and formulated by phase (grower 1, 25 to 50 and grower 2, 50 to 75 kg BW). The positive control diet (PC) provided adequate nutrients and a negative control diet (NC) was formulated without Pi (1.2 g/kg ATTD P) and reduced in Ca (-0.12 to -0.13 percentage points), NE (-32 kcal/kg), and digestible essential AA (-0.004 to -0.026 percentage points) vs. PC. Two further treatments comprised the NC plus 500 or 1,000 FTU/kg of PhyG. Data were analyzed by ANOVA, mean contrasts and orthogonal polynomial regression. Nutrient reductions in the NC reduced (P < 0.05) average daily gain (ADG) during grower 1 and overall (73 to 136 d of age), increased (P < 0.05) feed conversion ratio (FCR) during grower 1 and overall and tended to reduce (P < 0.1) average daily feed intake (ADFI) during grower 2 and overall, vs. PC. Phytase supplementation improved (P < 0.05) FCR during grower 1, ADG during grower 2 and overall, ATTD of DM and P, and tended to improve DE (P = 0.053) in a linear dose-dependent manner. PhyG at 1,000 FTU/kg resulted in growth performance (all measures, all phases) equivalent to PC. The findings demonstrate that PhyG at 1,000 FTU/kg totally replaced Pi in complex grower pig diets containing industrial co-products, compensated a full nutrient matrix reduction and maintained performance.

A novel consensus bacterial 6-phytase variant completely replaced inorganic phosphate in broiler diets, maintaining growth performance and bone quality: data from two independent trials.

Total replacement of dietary inorganic phosphate (Pi) by a novel consensus bacterial 6-phytase variant (PhyG) in phytate-rich diets (>0.3% phytate-P) was investigated in 2 trials using growth performance and bone quality as outcome measures. Both trials utilized a completely randomized design with 5 dietary treatments across 4 phases: starter (0-10 d), grower (10-21 d), finisher 1 (21-35 d), and finisher 2 (35-42 d). Treatments comprised a nutritionally adequate positive control (PC) diet containing monocalcium phosphate and 4 experimental diets (IPF1, IPF2, IPF3, and IPF4), all containing no added Pi and reduced in Ca by 0.2 to 0.3% units vs. PC. IPF1contained PhyG at 1,000 FTU/kg (all phases); IPF2 contained PhyG at 1,000 FTU/kg (all phases) and was additionally reduced in digestible AA, ME, and sodium (-0.2 to -0.4% points, -74 kcal/kg, -0.04% points, respectively, vs. PC); IPF3 contained PhyG at 3,000 FTU/kg in starter, 2,000 FTU/kg in grower, and 1,000 FTU/kg in finisher phases; and IPF4 contained xylanase (2,000 U/kg) and PhyG (2,000 FTU/kg in starter, 1,500 FTU/kg in grower, and 1,000 FTU/kg in finisher phases) and was additionally reduced in ME (-71 kcal/kg vs. PC). Ross 308 broilers were used (trial 1: n = 1,200 mixed sex; 24 birds per pen × 10 replicates; trial 2: n = 1,300 males; 26 birds × 10 replicates). During all phases in both trials, all IPF treatments maintained or improved BW, ADG, ADFI, FCR and BW-corrected FCRc and bone quality parameters vs. PC. vs. PC, treatment IPF3 increased ADG during starter phase (+10.8%) and reduced overall FCRc (-12 points, P < 0.05) in Trial 1, and increased overall ADG (+4.4%), day 35 and day 42 BW (+3.5%, +4.9%), and reduced overall FCRc (-11 points) in Trial 2 (P < 0.05). IPF4 produced equivalent performance to IPF3 (both trials). These are the first data to demonstrate total replacement of Pi by microbial phytase during an entire growth cycle in broiler diets.

The effects of coarse and wet feeding on performance parameters, gastrointestinal tract and tibia traits, and digesta phytase activity in egg-type pullets, either fed a low or moderate phosphorus diet.

In a 2 × 2 × 2 factorial design, the effects of dietary non-phytate phosphorus (NPP) levels, 0.17% (low) and 0.33% (moderate), diet moisture (dry and wet), and diet particle size (coarse and fine), were studied on egg production, characteristics of gastrointestinal tract (GIT) and tibia, digesta pH, and phytase activity in layer pullets (16 to 28 wk of age). The low NPP diet increased average daily water intake (ADWI) to ADFI ratio (4.2%) from 16 to 17 wk, but decreased this ratio (2.8%) from 23 to 27 wk. It decreased ADFI (1.5%) and egg mass production (3.8%) from 19 to 22 wk. It decreased egg weight (0.29 g) and ADWI (2.1%) from 23 to 27 wk. At 22 wk, the GIT relative empty organ weights were (g/kg BW) higher for proventriculus + gizzard (0.96), duodeneum (0.94), and jejunum + ileum (1.95) with the low vs. moderate NPP diet. The low NPP diet decreased digesta phytase activity in crop and proventriculus+gizzard at 28 wk. The wet diet increased ADFI, ADWI, and ADWI/ADFI ratio from 16 to 27 wk, egg mass production (3.0%) from 19 to 22 wk, and egg weight (0.45 g) from 23 to 27 wk. The wet diet also increased digesta phytase activity in proventriculus+gizzard. The coarse diet decreased ADFI from 19 to 22 wk (1.7%) and 23 to 27 wk (1.2%). The coarse diet caused reduced egg mass production (2.6%) from 23 to 27 wk. Egg shell breaking strength was increased on the coarse diet (0.9 Newton). The coarse diet increased ADWI/ADFI ratio from 16 to 27 wk, and increased relative gizzard weight by 1.95 and 0.81 g/kg BW at 22 and 28 wk, respectively. The coarse diet increased jejunal/ileal pH with 0.16 units at 28 wk. None of the tested parameters affected tibia characteristics. It was concluded that a low NPP diet did not clearly affect the studied parameters. The wet diet increased ADFI, ADWI, and egg production. The coarse diet increased ADWI, egg shell breaking strength, relative gizzard weight, and reduced ADFI and egg production.

Screening of freshwater and seawater microalgae strains in fully controlled photobioreactors for biodiesel production.

Strain selection is one of the primary hurdles facing cost-effective microalgal biodiesel production. Indeed, the strain used affects both upstream and downstream biodiesel production processes. This study presents a screening procedure that considers the most significant criteria in microalgal biodiesel production including TAG production and wet extraction and recovery of TAGs. Fourteen freshwater and seawater strains were investigated. Large variation was observed between the strains in all the screening criteria. The overall screening procedure ultimately led to the identification of Parachlorella kessleri UTEX2229 and Nannochloropsis gaditana CCMP527 as the best freshwater and seawater strains, respectively. They featured the largest areal TAG productivity equal to 2.7×10(-3) and 2.3×10(-3)kgm(-2)d(-1), respectively. These two strains also displayed encouraging cell fragility in a high pressure bead milling process with 69% and 98% cell disruption at 1750bar making them remarkable strains for TAG extraction in wet environment.

CHK1 expression in Gastric Cancer is modulated by p53 and RB1/E2F1: implications in chemo/radiotherapy response.

Radiation has a limited but relevant role in the adjuvant therapy of gastric cancer (GC) patients. Since Chk1 plays a critical function in cellular response to genotoxic agents, we aimed to analyze the role of Chk1 in GC as a biomarker for radiotherapy resistance. We analyzed Chk1 expression in AGS and MKN45 human GC cell lines by RT-QPCR and WB and in a small cohort of human patient's samples. We demonstrated that Chk1 overexpression specifically increases resistance to radiation in GC cells. Accordingly, abrogation of Chk1 activity with UCN-01 and its expression with shChk1 increased sensitivity to bleomycin and radiation. Furthermore, when we assessed Chk1 expression in human samples, we found a correlation between nuclear Chk1 accumulation and a decrease in progression free survival. Moreover, using a luciferase assay we found that Chk1's expression is controlled by p53 and RB/E2F1 at the transcriptional level. Additionally, we present preliminary data suggesting a posttranscriptional regulation mechanism, involving miR-195 and miR-503, which are inversely correlated with expression of Chk1 in radioresistant cells. In conclusion, Chk1/microRNA axis is involved in resistance to radiation in GC, and suggests Chk1 as a potential tool for optimal stratification of patients susceptible to receive adjuvant radiotherapy after surgery.

Optimization of bead milling parameters for the cell disruption of microalgae: process modeling and application to Porphyridium cruentum and Nannochloropsis oculata.

A study of cell disruption by bead milling for two microalgae, Nannochloropsis oculata and Porphyridium cruentum, was performed. Strains robustness was quantified by high-pressure disruption assays. The hydrodynamics in the bead mill grinding chamber was studied by Residence Time Distribution modeling. Operating parameters effects were analyzed and modeled in terms of stress intensities and stress number. RTD corresponded to a 2 CSTR in series model. First order kinetics cell disruption was modeled in consequence. Continuous bead milling was efficient for both strains disruption. SI-SN modeling was successfully adapted to microalgae. As predicted by high pressure assays, N. oculata was more resistant than P. cruentum. The critical stress intensity was twice more important for N. oculata than for P. cruentum. SI-SN modeling allows the determination of operating parameters minimizing energy consumption and gives a scalable approach to develop and optimize microalgal disruption by bead milling.

Effects of Xylo-Oligosaccharides on Broiler Chicken Performance and Microbiota.

In broiler chickens, feed additives, including prebiotics, are widely used to improve gut health and to stimulate performance. Xylo-oligosaccharides (XOS) are hydrolytic degradation products of arabinoxylans that can be fermented by the gut microbiota. In the current study, we aimed to analyze the prebiotic properties of XOS when added to the broiler diet. Administration of XOS to chickens, in addition to a wheat-rye-based diet, significantly improved the feed conversion ratio. XOS significantly increased villus length in the ileum. It also significantly increased numbers of lactobacilli in the colon and Clostridium cluster XIVa in the ceca. Moreover, the number of gene copies encoding the key bacterial enzyme for butyrate production, butyryl-coenzyme A (butyryl-CoA):acetate CoA transferase, was significantly increased in the ceca of chickens administered XOS. In this group of chickens, at the species level, Lactobacillus crispatus and Anaerostipes butyraticus were significantly increased in abundance in the colon and cecum, respectively. In vitro fermentation of XOS revealed cross-feeding between L. crispatus and A. butyraticus. Lactate, produced by L. crispatus during XOS fermentation, was utilized by the butyrate-producing Anaerostipes species. These data show the beneficial effects of XOS on broiler performance when added to the feed, which potentially can be explained by stimulation of butyrate-producing bacteria through cross-feeding of lactate and subsequent effects of butyrate on gastrointestinal function.

Mad2 and BubR1 modulates tumourigenesis and paclitaxel response in MKN45 gastric cancer cells.

Aneuploidy and chromosomal instability (CIN) are common features of gastric cancer (GC), but their contribution to carcinogenesis and antitumour therapy response is still poorly understood. Failures in the mitotic checkpoint induced by changes in expression levels of the spindle assembly checkpoint (SAC) proteins cause the missegregation of chromosomes in mitosis as well as aneuploidy. To evaluate the possible contribution of SAC to GC, we analyzed the expression levels of proteins of the mitotic checkpoint complex in a cohort of GC cell lines. We found that the central SAC proteins, Mad2 and BubR1, were the more prominently expressed members in disseminated GC cell lines. Silencing of Mad2 and BubR1 in MKN45 and ST2957 cells decreased their cell proliferation, migration and invasion abilities, indicating that Mad2 and BubR1 could contribute to cellular transformation and tumor progression in GC. We next evaluated whether silencing of SAC proteins could affect the response to microtubule poisons. We discovered that paclitaxel treatment increased cell survival in MKN45 cells interfered for Mad2 or BubR1 expression. However, apoptosis (assessed by caspase-3 activation, PARP proteolysis and levels of antiapoptotic Bcl 2-family members), the DNA damage response (assessed by H2Ax phosphorylation) and exit from mitosis (assessed by Cyclin B degradation and Cdk1 regulation) were activated equally between cells, independently of Mad2 or BubR1-protein levels. In contrast, we observed that the silencing of Mad2 or BubR1 in MKN45 cells showed the induction of a senescence-like phenotype accompanied by cell enlargement, increased senescence-associated ß-galactosidase activity and increased IL-6 and IL-8 expression. In addition, the senescent phenotype is highly increased after treatment with PTX, indicating that senescence could prevent tumorigenesis in GC. In conclusion, the results presented here suggest that Mad2 and BubR1 could be used as prognostic markers of tumor progression and new pharmacological targets in the treatment for GC.

Selective extraction from microalgae Nannochloropsis sp. using different methods of cell disruption.

This work studies the extraction of intracellular components from microalgae Nannochloropsis sp. with application of different cell disruption techniques, including pulsed electric field (PEF) (20kV/cm, 1-4ms, 13.3-53.1kJ/kg), high voltage electrical discharge (HVED) (40kV/cm, 1-4ms, 13.3-53.1kJ/kg), ultrasonication (USN) (200W, 1-8min, 12-96kJ/kg), and high pressure homogenization (HPH) (150MPa, 1-10 passes, 150-1500kJ/kg). The data evidence that electrically based disruption techniques (PEF and HVED) allowed selective extraction of water soluble ionic components and microelements, small molecular weight organic compounds and water soluble proteins. Microscopic and sedimentation stability analyses have shown that microalgae cells in HVED-treated suspension were noticeably agglomerated and could be easily settled in centrifuge. The electrically based disruption techniques were ineffective for delivery of pigments (e.g., chlorophylls or carotenoids) and their extraction required subsequent application of more potent disruption techniques. The obtained data have shown that HPH disruption technique was the most effective; however, this mode required the highest power consumption.

Interest of second harmonic generation imaging for diagnosis in thick and opaque tissue.

In articular hyaline cartilage, chondrocytes are surrounded by an extracellular matrix which is mainly composed by collagen and proteoglycanes. Pathological specimens show a partial or complete degradation of this matrix. Therefore, it could be interesting to know how mechanical or biochemical constraints applied to cartilage specimens induce modifications of the cartilage network. Multiphoton technology combined to Second Harmonic Generation (SHG) enables to image cartilage specimens in a non-invasive mode with high resolution at deep penetration. By placing a band pass filter in front of the transmitted light detector, SHG signal with frequency doubled can be isolated for a new contrast imaging. SHG (second harmonic generation) is a diffusion process generated from organized structures and does not need any fluorescent staining. Due to their non-centrosymetric structure, collagen fibrilles present a high second-order non-linear susceptibility and thus give rise to a strong SHG signal when exposed to high enough electric fields produced by a focal point of a femtosecond pulsed laser (multiphoton microscopy). As the extracellular matrix of cartilage is in part constituted by collagen fibers, it can be imaged with this contrast tool. The intensity of SHG signals strongly depends on the organization of collagen fibers. Thus a modification of the extracellular matrix in terms of 3D-organization of collagen induced by mechanical stress can be shown with this contrast tool.

Influences of construct properties on the proliferation and matrix synthesis of dedifferentiated chondrocytes cultured in alginate gel.

To investigate whether the chondrocytes-alginate construct properties, such as cell seeding density and alginate concentration might affect the redifferentiation, dedifferentiated rat articular chondrocytes were encapsulated at low density (LD: 3 x 10(6) cells/ml) or high density (HD: 10 x 10(6) cells/ml) in two different concentrations of alginate gel (1.2% or 2%, w/v) to induce redifferentiation. Cell viability and cell proliferation of LD culture was higher than those of HD culture. The increase in alginate gel concentration did not make an obvious difference in cell viability, but reduced cell proliferation rate accompanied with the decrease of cell population in S phase and G2/M phase. Scan electron microscopy observation revealed that chondrocytes maintained round in shape and several direct cell-cell contacts were noted in HD culture. In addition, more extracellular matrix was observed in the pericellular region of chondrocytes in 2% alginate culture than those in 1.2% alginate culture. The same tendency was found for the synthesis of collagen type II. No noticeable expression of collagen type I was detected in all constructs at the end of 28-day cultures. These results suggested that construct properties play an important role in the process of chondrocytes' redifferentiation and should be considered for creating of an appropriate engineered articular cartilage.

Multiphoton microscopy for blood vessel imaging: new non-invasive tools (Spectral, SHG, FLIM).

Imaging thick and opaque tissue, like blood vessel, in a noninvasive mode with high resolution, is nowadays possible with multiphoton technology. A near-infrared excitation presents the advantage to be compatible with living specimens and allows a deep penetration into tissues. The nonlinear excitation process is followed by several deactivation ways, among which fluorescence emission can be represented with Spectral or Lifetime imaging. Applied to ex vivo blood vessel imaging, these techniques enabled us to discriminate cell structures (nucleus, cytoskeleton) by fluorescent labelling (Hoechst, QDots). Another method, based on 2-photon excitation and which doesn't need any exogenous dye has also been experimented on arteries: SHG (Second Harmonic Generation) is a diffusion process generated from organized structures. Collagen molecules give rise to a strong SHG signal, enabling us to image the arterial wall (3-dimensional extracellular matrix).

Decellularized umbilical artery treated with thin polyelectrolyte multilayer films: potential use in vascular engineering.

Decellularized allograft tissues have been identified as a potential extracellular matrix scaffold for tissue-engineered vascular substitutes. In order to improve the thromboresistance, it is necessary to pre-coat the intra-luminal vessel surface. Recently a new surface modification technique appeared, based on the alternate adsorption of positive and negative charged polyelectrolytes. Our objective was to develop an alternative vascular scaffold made of decellularized human umbilical arteries treated with a PAH/PSS polyelectrolyte multilayered film. The vessels luminal surfaces covered with the multilayer film were observed by electronic scanning microscopy. Our observations showed that the luminal surface is completely devoid of ECs following treatment with trypsin. A top view of the coated artery indicated that the multilayer uniformly covered internal surface of the vessels. The successful of the multilayer correct deposition and retention on the arterial wall were controlled by confocal microscopy using a fluorescent polyelectrolyte (rhodamine-PAH). The data suggest that decellularized cryopreserved arteries represent a potential scaffold for further vascular tissue engineering efforts. Moreover, the multilayer films can be used to coat biological surfaces and following the terminated layer (PAH or PSS), favour the cell adhesion or cell resistance.

The risk of post-operative complications associated with infliximab therapy for Crohn's disease: a controlled cohort study.

BACKGROUND: By temporarily suppressing the immune response, the anti-tumour necrosis factor agent, infliximab, may increase the risk of peri-operative complications. AIM: To test this hypothesis for intestinal resection in a cohort of 313 Crohn's disease patients treated with infliximab. Forty received one or more infusions prior to intestinal resection (31/40 within 12 weeks). METHODS: The post-operative events of these patients were compared with those of a control group (infliximab naive) of 39 patients adjusted for age, gender and surgical procedure. Early (10 days) and late (3 months) major or minor complications were identified. RESULTS: The incidence of early minor (15.0% vs. 12.8%) and major (12.5% vs. 7.7%) and late minor (2.5% vs. 5.1%) and major (17.5% vs. 12.8%) complications and the mean hospital stay after surgery (10.3 +/- 4.0 days vs. 9.9 +/- 5.5 days) were similar in both groups. A trend towards an increased early infection rate was found in infliximab pre-treated patients (6 vs. 1; P = 0.10), but more patients in this group received corticosteroids and/or immunosuppressives (29 vs. 16 patients; P < 0.05). CONCLUSION: The use of infliximab before intestinal resection does not prolong the hospital stay and does not increase the rate of post-operative complications.