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See detailIn vitro evaluation of biocompatibility of uncoated thermally reduced graphene and carbon nanotube-loaded PVDF membranes with adult neural stem cell-derived neurons and glia
Defteralı, Çağla; Verdejo, Raquel; Majeed, Shahid et al

in Frontiers in Bioengineering and Biotechnology (2016), 4(n° 64),

Graphene, graphene-based nanomaterials (GBNs), and carbon nanotubes (CNTs) are being investigated as potential substrates for the growth of neural cells. However, in most in vitro studies, the cells were ... [more ▼]

Graphene, graphene-based nanomaterials (GBNs), and carbon nanotubes (CNTs) are being investigated as potential substrates for the growth of neural cells. However, in most in vitro studies, the cells were seeded on these materials coated with various proteins implying that the observed effects on the cells could not solely be attributed to the GBN and CNT properties. Here, we studied the biocompatibility of uncoated thermally reduced graphene (TRG) and poly(vinylidene fluoride) (PVDF) membranes loaded with multi-walled CNTs (MWCNTs) using neural stem cells isolated from the adult mouse olfactory bulb (termed aOBSCs). When aOBSCs were induced to differentiate on coverslips treated with TRG or control materials (polyethyleneimine-PEI and polyornithine plus fibronectin- PLO/F) in a serum-free medium, neurons, astrocytes, and oligodendrocytes were generated in all conditions, indicating that TRG permits the multi-lineage differentiation of aOBSCs. However, the total number of cells was reduced on both PEI and TRG. In a serum-containing medium, aOBSC-derived neurons and oligodendrocytes grown on TRG were more numerous than in controls; the neurons developed synaptic boutons and oligodendrocytes were more branched. In contrast, neurons growing on PVDF membranes had reduced neurite branching, and on MWCNTs-loaded membranes oligodendrocytes were lower in numbers than in controls. Overall, these findings indicate that uncoated TRG may be biocompatible with the generation, differentiation, and maturation of aOBSC-derived neurons and glial cells, implying a potential use for TRG to study functional neuronal networks. [less ▲]

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See detailIn vitro evaluation of fermentation characteristics of two types of insects as potential novel protein feeds for pigs
POELAERT, Christine ULg; Beckers, Yves ULg; Despret, Xavier et al

in Journal of Animal Science (2016), 94(7S3), 198-201

Novel protein sources such as insects are suggested for pig nutrition. Protein availability might be impacted by the nature of the insect and by the thermal treatment applied to sanitize this ingredient ... [more ▼]

Novel protein sources such as insects are suggested for pig nutrition. Protein availability might be impacted by the nature of the insect and by the thermal treatment applied to sanitize this ingredient. Their influence on protein availability and colonic fermentation is unknown. Plant proteins (beans, lentils, peas, and soybean, raw and vapor cooked) were compared to adult house crickets (Acheta domesticus) and mealworm larvae (Tenebrio molitor) that had been autoclaved, oven cooked (150 and 200°C), or used raw. Ingredients were run in an in vitro model of the pig gastrointestinal tract combining enzymes to simulate digestion in the stomach and the small intestine and subsequent fermentation by fecal microbes to simulate hindgut fermentation. In vitro crude protein disappearance (IVCPD) of insects decreased with oven cooking at 150°C or autoclaving (P < 0.05) while that of plants was unaffected (P > 0.05), except for soybean. IVCPD of raw mealworms (0.726) equaled that of the best plants (0.725 to 0.763) while crickets were less digestible (P < 0.01). Consequences on fermentation metabolites were lower propionate (P < 0.01) and branched-chain fatty acids (BCFA; P < 0.05) molar ratio in raw insects against oven-cooked or autoclaved insects. Both insect sources displayed greater BCFA (P < 0.01) and lower propionate (P < 0.01) than plants. Crickets produced 50% as much BCFA as mealworms (P < 0.01). In conclusion, feeding insect-sourced protein requires a careful choice of the species as well as the thermal treatment to avoid possible detrimental consequences on digestibility and intestinal health in pigs. [less ▲]

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See detailIn vitro evaluation of fermentation characteristics of two types of insects, as potential novel protein feeds for pigs
POELAERT, Christine ULg; Beckers, Yves ULg; Despret, Xavier et al

in 13th International Symposium on Digestive Physiology of Pigs (2015, May)

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See detailIn vitro evaluation of glutathione peroxidase (GPx)-like activity and antioxidant properties of some Ebselen analogues
Mouithys-Mickalad, Ange ULg; Mareque-Faez, Juan; Chistiaens, U. et al

in Redox Report : Communications in Free Radical Research (2004), 9(2), 81-87

Four analogues of Ebselen were synthesized and their glutathione peroxidase activity and antioxidant property evaluated and compared to Ebselen. Among the studied compounds, only diselenide [3] exhibited ... [more ▼]

Four analogues of Ebselen were synthesized and their glutathione peroxidase activity and antioxidant property evaluated and compared to Ebselen. Among the studied compounds, only diselenide [3] exhibited both glutathione peroxidase activity and radical-scavenging capability. Compounds [3] and [4] showed a strong inhibitory effect (53% and 43%, respectively) on the lipid peroxidation of linoleic acid compared to Ebselen and selenide derivatives ([1] and [2]) which were less active (28%, 26% and 18% inhibition, respectively). A concentration-dependent inhibitory effect was also found in the model of the formation of ABTS*+ radical cation: 65% and 89% inhibition for compound [3] at 10(-4) M and 5 x 10(-5) M, respectively, and 68% and 90% for compound [4], compared to 14% and 52% inhibition for Ebselen and the diselenides [1] and [2] (29%, 46% and 45%, 68%, respectively). By EPR spin trapping technique, the following inhibitory profile of the Ebselen analogues was observed towards the formation of thiyl radicals: Ebselen = [3]>[1]>[2]>[4]. Studies with compound [3] are in progress on oxidative stress cell models. [less ▲]

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See detailAn in vitro evaluation of leakage of two Etch and Rinse and two Self-Etch adhesives after thermocycling
Geerts, Sabine ULg; BOLETTE, Amandine ULg; SEIDEL, Laurence ULg et al

in International Journal of Dentistry (2012)

Our experiment evaluated the microleakage in resin composite restorations bonded to dental tissues with different adhesive systems. 40 class V cavities were prepared on the facial and lingual surfaces of ... [more ▼]

Our experiment evaluated the microleakage in resin composite restorations bonded to dental tissues with different adhesive systems. 40 class V cavities were prepared on the facial and lingual surfaces of each tooth with coronal margins in enamel and apical margins in cementum (root dentin). The teeth were restored with Z100 resin composite bonded with different adhesive systems: Scotchbond! Multipurpose (SBMP) a 3-step Etch and Rinse adhesive, Adper! Scotchbond! 1 XT (SB1) a 2-step Etch and Rinse adhesive, AdheSE® One (ADSE-1) a 1-step Self-Etch adhesive and AdheSE® (ADSE) a 2-step Self-Etch adhesive. Teeth were thermocycled and immersed in 50 % silver nitrate solution. When both interfaces were considered, SBMP has exhibited significantly less microleakage than other adhesive systems (respectively for SB1, ADSE-1 and ADSE, p = 0.0007, p < 0.0001 and p < 0.0001). When enamel and dentin interfaces were evaluated separately: 1) for the Self-Etch adhesives, microleakage was found greater at enamel than at dentin interfaces (for ADSE, p = 0.024 and for ADSE-1, p < 0.0001); 2) for the Etch and Rinse adhesive systems, there was no significant difference between enamel and dentin interfaces; 3) SBMP was found significantly better than other adhesives both at enamel and dentin interfaces. In our experiment Etch and Rinse adhesives remain better than Self-Etch adhesives at enamel interface. In addition, there was no statistical difference between 1-step (ADSE-1) and 2-step (ADSE) Self-Etch adhesives. [less ▲]

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See detailIn vitro evaluation of lipid matrices for the development of a sustained-release sulfamethazine bolus for lambs
Evrard, Brigitte ULg; Delattre, Luc ULg

in Drug Development & Industrial Pharmacy (1996), 22(2), 111-118

The aim of this study is the development of a high-density bolus to be given to lambs in order to release sulfamethazine during 4 days after a single oral administration. The suitability of 12 lipid ... [more ▼]

The aim of this study is the development of a high-density bolus to be given to lambs in order to release sulfamethazine during 4 days after a single oral administration. The suitability of 12 lipid matrix formulations was assessed in order to maintain the integrity of the tablet even after complete dissolution of the active substance. The influence of both nature and concentration of the lipid excipient on the in vitro release of sulfamethazine was investigated. The influences of the granulation method and of the compression force were assessed on a formulation containing Cutina® HR as a binding and sustained-release agent. Finally, stability tests showed that the in vitro release characteristics remain unchanged during storage. [less ▲]

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See detailIn vitro evaluation of protein precipitation capacity of temperate browse species
Vandermeulen, Sophie ULg; Leblois, Julie ULg; Ramírez-Restrepo, Carlos Alberto et al

Poster (2016, February 05)

European agri-environmental policies are promoting the establishment of shrubs and trees on grasslands. The use of browse as fodder requires knowledge on their nutritive value since intensive production ... [more ▼]

European agri-environmental policies are promoting the establishment of shrubs and trees on grasslands. The use of browse as fodder requires knowledge on their nutritive value since intensive production systems are still relying on expensive and environment-costing protein sources. However, information on the influence of temperate condensed tannins (CT)-containing browse forage on rumen protein metabolism is elusive. The study aimed to assess the protein precipitation capacity (PPC) of 10 temperate browse species and establish the correlation between PPC values and plants CT content. PPC of foliage of 3 individuals per woody plants was measured using 2 model proteins: bovine serum albumin (BSA) and casein. The N content in protein solutions (4.6g/L; pH=6.8) was determined before and after adding each forage sample. Extractable CT concentration was quantified by spectrophotometry. The PPC varied across plant species (P<0.001). Corylus avellana had the highest ability to precipitate casein (52.4%). In contrast, the BSA precipitation (18.3%) of this plant was similar to Cornus sanguinea (12.7%), Quercus robur (12.1%) and Crataegus monogyna (11.0%). CT content ranged from 1.4 in Fraxinus excelsior to 82.7g/kg of depigmented sample in Corylus avellana (P<0.001) and was correlated to BSA (r=0.70; P<0.001) and casein PC (r=0.51; P<0.01). It was concluded that woody species could play a significant role in modifying protein metabolism, but further in vivo trials are required. [less ▲]

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See detailIn vitro evaluation of the anti-apoptotic drug Z-VAD-FMK on human ovarian granulosa cell lines for further use in ovarian tissue transplantation.
Fransolet, Maïté ULg; HENRY, Laurie ULg; Labied, Soraya et al

in Journal of Assisted Reproduction & Genetics (2015)

PURPOSE: Because ovarian granulosa cells are essential for oocyte survival, we examined three human granulosa cell lines as models to evaluate the ability of the pan-caspase inhibitor benzyloxycarbonyl ... [more ▼]

PURPOSE: Because ovarian granulosa cells are essential for oocyte survival, we examined three human granulosa cell lines as models to evaluate the ability of the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (Z-VAD-FMK) to prevent primordial follicle loss after ovarian tissue transplantation. METHODS: To validate the efficacy of Z-VAD-FMK, three human granulosa cell lines (GC1a, HGL5, COV434) were treated for 48 h with etoposide (50 mug/ml) and/or Z-VAD-FMK (50 muM) under normoxic conditions. To mimic the ischemic phase that occurs after ovarian fragment transplantation, cells were cultured without serum under hypoxia (1 % O2) and treated with Z-VAD-FMK. The metabolic activity of the cells was evaluated by WST-1 assay. Cell viability was determined by FACS analyses. The expression of apoptosis-related molecules was assessed by RT-qPCR and Western blot analyses. RESULTS: Our assessment of metabolic activity and FACS analyses in the normoxic experiments indicate that Z-VAD-FMK protects granulosa cells from etoposide-induced cell death. When cells are exposed to hypoxia and serum starvation, their metabolic activity is reduced. However, Z-VAD-FMK does not provide a protective effect. In the hypoxic experiments, the number of viable cells was not modulated, and we did not observe any modifications in the expressions of apoptosis-related molecules (p53, Bax, Bcl-xl, and poly (ADP-ribose) polymerase (PARP)). CONCLUSION: The death of granulosa cell lines was not induced in our ischemic model. Therefore, a protective effect of Z-VAD-FMK in vitro for further use in ovarian tissue transplantation could not be directly confirmed. It will be of interest to potentially use Z-VAD-FMK in vivo in xenograft models. [less ▲]

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See detailIn vitro evaluation of the fermentation characteristics in the pig intestines of hulless barleys differing in β-glucan content
Jha, Rajesh; Bindelle, Jérôme ULg; Rossnagel, Brian et al

in Journal of Animal Science (2009), 87(E-Suppl. 3), 103

Non-starch polysaccharides (NSP) in isolated form, especially β-glucans, are reported to have prebiotic effects in pigs. However, little information is available on the possible functional properties of ... [more ▼]

Non-starch polysaccharides (NSP) in isolated form, especially β-glucans, are reported to have prebiotic effects in pigs. However, little information is available on the possible functional properties of these NSP when the latter are still present in the fibrous matrix of whole cereals. Hulless barleys (HB) are good sources of β-glucans and the content is quite variable among varieties. In order to evaluate the potential of HB as functional feeds, an in vitro experiment was carried out to study the fermentation characteristics of 6 HB varieties varying in their β-glucan contents (36-99 g/kg DM) in comparison to 3 hulled barleys and 5 oats. After a pepsin-pancreatin hydrolysis, the ingredients were incubated in a buffer solution containing minerals and pig feces as inoculum. The accumulated gas production, proportional to the amount of fiber fermented, was measured for 48 h and modeled. Short-chain fatty acid (SCFA) and ammonia concentration were measured in the fermented solutions. A cereal type effect (P < 0.05) was observed on the fermentation kinetics parameters. Rates of degradation and total gas productions were higher in HB than in oats (P < 0.05) but no difference was observed between HB varieties. On the contrary, differences were found between HB for lag time and rate of degradation. The production of SCFA was also higher with HB (6.1 mMol/g DM incubated; P < 0.05) than with hulled barleys and oats (4.9 and 2.9 mMol/g DM incubated respectively). In contrast, oats generated higher ammonia (P <0.05) production (1.4 mMol/g DM incubated, on average) than barley (1.0 mMol/g). In conclusion, HB are better fermented in vitro, produce more beneficial (SCFA) and less harmful (ammonia) metabolites and have a better potential than other cereal species to modulate gut microbiota and improve gut health. [less ▲]

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See detailIn vitro evaluation of the fermentation characteristics of the carbohydrate fractions of hulless barley and other cereals in the gastrointestinal tract of pigs
Jha, Rajesh; Bindelle, Jérôme ULg; Rossnagel, Brian et al

in Animal Feed Science & Technology (2011), 163

An in vitro model was used to study the fermentation characteristics of carbohydrate fractions of hulless barley (hB), in comparison to hulled barley (HB), hulled oat and oat groats (OG) in the pig ... [more ▼]

An in vitro model was used to study the fermentation characteristics of carbohydrate fractions of hulless barley (hB), in comparison to hulled barley (HB), hulled oat and oat groats (OG) in the pig intestine. For this purpose, 6 hulless barley cultivars (hB), varying in β-glucan content (36-99 g/kg DM), were compared to 3 HB cultivars, 2 oat groat samples (OG), 3 oat varieties and a reference sample of wheat. The residue of a pepsin-pancreatin hydrolysis was incubated in a buffered mineral solution inoculated with pig faeces. Gas production, proportional to the amount of fermented carbohydrates, was measured for 48 h and kinetics modelled. The fermented solution was subsequently analyzed for microbial production of short-chain fatty acids (SCFA) and ammonia. In vitro dry matter degradability varied according to ingredient (P<0.001). Higher values were observed for OG, ranging from 0.88 to 0.99 as compared to oat, hB and HB, for which degradability ranged from 0.63 to 0.73, 0.68 to 0.80 and 0.69 to 0.71, respectively. A “cereal type” effect (P<0.05) was observed on fermentation kinetics parameters. Total gas production was higher (P<0.05) with hB (224 ml/g DM incubated) than with HB and oat (188 and 55 ml/g DM incubated, respectively). No difference was observed between hB cultivars (P>0.05) for total gas production but differences (P<0.001) were found for lag time and the fractional rate of degradation. Hulless barley cultivar CDC Fibar (waxy starch) and CDC McGwire (normal starch) started to ferment sooner (lag time of 0.7 and 0.9 h, respectively) than SH99250 (high amylose starch; 1.7 h). The fractional rate of degradation was similar in both hB and OG (0.15/h on average), which was higher than that of HB (0.12/h). The production of SCFA was also higher (P<0.05) with hB (6.1 mmol/g DM incubated, on average) than with HB and oat (4.9 and 2.9 mmol/g DM incubated, respectively). Similar trends were found for SCFA production expressed per g fermented carbohydrates, with higher butyrate and lower acetate ratio. In contrast, oat fermentation generated higher (P<0.05) ammonia concentration (1.4 mmol/g DM incubated, on average) than hB (1.0 mmol/g DM incubated). In summary, hulless barleys, irrespective of cultivar type had higher in vitro fermentability and produced more SCFA and less ammonia than hulled barley and oat. Thus, hulless barleys have a better potential to be used in pig nutrition to manipulate the fermentation activity in the intestine of pigs. [less ▲]

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See detailIn vitro expression of RC2-labeled antigens as radial glial cells markers in cultures derived from mouse embryonic brain
Leprince, P; Chanas-Sacré, G; Lewin, M et al

Poster (1996)

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See detailIn vitro fibre fermentation characteristics of specialty ingredients with varying NSP levels
Jha, Rajesh; Bindelle, Jérôme ULg; Rossnagel, Brian et al

Poster (2009)

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See detailIn vitro fibre fermentation of feed ingredients with varying fermentable carbohydrate and protein levels and protein synthesis by colonic bacteria isolated from pigs
Jha, Rajesh; Bindelle, Jérôme ULg; Van Kessel, Andrew et al

in Animal Feed Science & Technology (2011), 165

An in vitro experiment was carried out using the gas technique to study the fermentation characteristics of different feed ingredients differing in their fermentable carbohydrate and protein composition ... [more ▼]

An in vitro experiment was carried out using the gas technique to study the fermentation characteristics of different feed ingredients differing in their fermentable carbohydrate and protein composition by colonic bacteria isolated from pigs. The effect on in vitro bacterial protein synthesis was also evaluated. The ingredients used were wheat bran (WB), wood cellulose (Solka-floc®, SF), peas, pea hulls (PH), pea inner fibre (PIF), sugar beet pulp (SBP), flax seed meal (FSM) and corn distillers dried grains with solubles (DDGS). The samples were pre-treated with pepsin and pancreatin and the hydrolyzed substrates were then incubated with pig faeces in a buffered mineral solution. The nitrogen source in the buffer solution (NH4HCO3) was replaced by an equimolar quantity of 15N-labeled NH4Cl, used for the determination of the rate of bacterial protein synthesis. Gas production, proportional to the amount of fermented carbohydrate, was recorded for 48 h and modelled. The fermented product was subjected to short-chain fatty acids (SCFA) analysis. The source of fibre affected the in vitro dry matter degradability (IVDMD), the fermentation kinetics and the gas production profile (P<0.05). The highest (P<0.001) IVDMD values were observed for peas (0.80) and FSM (0.70), whereas SF was essentially undegraded (0.06). The fractional rate of degradation appeared to be lower (P<0.001) for WB and DDGS (0.07 and 0.05 h, respectively) and highest for SBP (0.20 h). Peas started to ferment rapidly (lag time 1.3 h). Half gas production (T/2) was achieved sooner for PIF (8.4 h) and was the longest for DDGS (19.8 h). The total gas production was the highest for PH, followed by SF, PIF and peas (276, 266, 264 and 253 ml/g DM incubated, respectively) and the lowest for FSM and WB (130 and 124 ml/g DM incubated, respectively). There was no difference (P>0.05) in SCFA production after the fermentation of SF, P, PH, PIF and SBP (ranging from 3.8 to 4.5 mmol/g DM incubated) while WB and FSM yielded lowest (P<0.05) SCFA. The bacterial nitrogen incorporation (BNI), both at T/2 and after 48 h of fermentation was the highest (P<0.001) for PIF (18.5 and 15.6 mg/g DM incubated, respectively) and the lowest for DDGS and WB. In conclusion, peas and pea fibres had higher rates of fermentability, produced more SCFA and had high bacterial protein synthesis capacity. They thus have the potential to be included in pig diets as a source of fermentable fibre to modulate the gut environment and reduce nitrogen excretion. [less ▲]

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