XIAP downregulation accompanies mebendazole growth inhibition in melanoma xenografts

Mebendazole (MBZ) was identified as a promising therapeutic on the basis of its ability to induce apoptosis in melanoma cell lines through a B-cell lymphoma 2 (BCL2)-dependent mechanism. We now show that in a human xenograft melanoma model, oral MBZ is as effective as the current standard of care temozolomide in reducing tumor growth. Inhibition of melanoma growth in vivo is accompanied by phosphorylation of BCL2 and decreased levels of X-linked inhibitor of apoptosis (XIAP). Reduced expression of XIAP on treatment with MBZ is partially mediated by its proteasomal degradation. Furthermore, exposure of melanoma cells to MBZ promotes the interaction of SMAC/DIABLO with XIAP, thereby alleviating XIAP's inhibition on apoptosis. XIAP expression on exposure to MBZ is indicative of sensitivity to MBZ as MBZ-resistant cells do not show reduced levels of XIAP after treatment. Resistance to MBZ can be reversed partially by siRNA knockdown of cellular levels of XIAP. Our data indicate that MBZ is a promising antimelanoma agent on the basis of its effects on key antiapoptotic proteins.

Viable chimaeric viruses confirm the biological importance of sequence specific maize streak virus movement protein and coat protein interactions

Background. A variety of interactions between up to three different movement proteins (MPs), the coat protein (CP) and genomic DNA mediate the inter- and intra-cellular movement of geminiviruses in the genus Begomovirus. Although movement of viruses in the genus Mastrevirus is less well characterized, direct interactions between a single MP and the CP of these viruses is also clearly involved in both intra- and intercellular trafficking of virus genomic DNA. However, it is currently unknown how specific these MP-CP interactions are, nor how disruption of these interactions might impact on virus viability. Results. Using chimaeric genomes of two strains of Maize streak virus (MSV) we adopted a genetic approach to investigate the gross biological effects of interfering with interactions between virus MP and CP homologues derived from genetically distinct MSV isolates. MP and CP genes were reciprocally exchanged, individually and in pairs, between maize (MSV-Kom)- and Setaria sp. (MSV-Set)-adapted isolates sharing 78% genome-wide sequence identity. All chimaeras were infectious in Zea mays c.v. Jubilee and were characterized in terms of symptomatology and infection efficiency. Compared with their parental viruses, all the chimaeras were attenuated in symptom severity, infection efficiency, and the rate at which symptoms appeared. The exchange of individual MP and CP genes resulted in lower infection efficiency and reduced symptom severity in comparison with exchanges of matched MP-CP pairs. Conclusion. Specific interactions between the mastrevirus MP and CP genes themselves and/or their expression products are important determinants of infection efficiency, rate of symptom development and symptom severity. © 2008 van der Walt et al; licensee BioMed Central Ltd.

HIV-1 sub-type C chimaeric VLPs boost cellular immune responses in mice

Several approaches have been explored to eradicate HIV; however, a multigene vaccine appears to be the best option, given their proven potential to elicit broad, effective responses in animal models. The Pr55 Gagprotein is an excellent vaccine candidate in its own right, given that it can assemble into large, enveloped, virus-like particles (VLPs) which are highly immunogenic, and can moreover be used as a scaffold for the presentation of other large non-structural HIV antigens. In this study, we evaluated the potential of two novel chimaeric HIV-1 Pr55 Gag-based VLP constructs - C-terminal fusions with reverse transcriptase and a Tat::Nef fusion protein, designated GagRT and GagTN respectively - to enhance a cellular response in mice when used as boost components in two types of heterologous prime-boost vaccine strategies. A vaccine regimen consisting of a DNA prime and chimaeric HIV-1 VLP boosts in mice induced strong, broad cellular immune responses at an optimum dose of 100 ng VLPs. The enhanced cellular responses induced by the DNA prime-VLP boost were two- to three-fold greater than two DNA vaccinations. Moreover, a mixture of GagRT and GagTN VLPs also boosted antigen-specific CD8+ and CD4+ T-cell responses, while VLP vaccinations only induced predominantly robust Gag CD4+ T-cell responses. The results demonstrate the promising potential of these chimaeric VLPs as vaccine candidates against HIV-1. © 2010 Pillay et al; licensee BioMed Central Ltd.

Effects of acute multinutrient supplementation on rugby union game performance and recovery

Purpose: To investigate the effects of an acute multinutrient supplement on game-based running performance, peak power output, anaerobic by-products, hormonal profiles, markers of muscle damage, and perceived muscular soreness before, immediately after, and 24 h following competitive rugby union games. Methods: Twelve male rugby union players ingested either a comprehensive multinutrient supplement (SUPP), [RE-ACTIVATE:01], or a placebo (PL) for 5 d. Participants then performed a competitive rugby union game (with global positioning system tracking), with associated blood draws and vertical jump assessments pre, immediately post and 24 h following competition. Results: SUPP ingestion resulted in moderate to large effects for augmented 1st half very high intensity running (VHIR) mean speed (5.9 ± 0.4 vs 4.8 ± 2.3 m·min–1; d= 0.93). Further, moderate increases in 2nd half VHIR distance (137 ± 119 vs 83 ± 89 m; d= 0.73) and VHIR mean speed (5.9 ± 0.6 v 5.3 ± 1.7 m·min–1; d= 0.56) in SUPP condition were also apparent. Postgame aspartate aminotransferase (AST; 44.1 ± 11.8 vs 37.0 ± 3.2 UL; d= 1.16) and creatine kinase (CK; 882 ± 472 vs. 645 ± 123 UL; d= 0.97) measures demonstrated increased values in the SUPP condition, while AST and CK values correlated with 2nd half VHIR distance (r= –0.71 and r= –0.76 respectively). Elevated C-reactive protein (CRP) was observed postgame in both conditions; however, it was significantly blunted with SUPP (P= .05). Conclusions: These findings suggest SUPP may assist in the maintenance of VHIR during rugby union games, possibly via the buffering qualities of SUPP ingredients. However, correlations between increased work completed at very high intensities and muscular degradation in SUPP conditions, may mask any anticatabolic properties of the supplement.

Effects of mixed-method cooling on recovery of medium-fast bowling performance in hot conditions on consecutive days

This investigation examined physiological and performance effects of cooling on recovery of medium-fast bowlers in the heat. Eight, medium-fast bowlers completed two randomised trials, involving two sessions completed on consecutive days (Session 1: 10-overs and Session 2: 4-overs) in 31 ± 3°C and 55 ± 17% relative humidity. Recovery interventions were administered for 20 min (mixed-method cooling vs. control) after Session 1. Measures included bowling performance (ball speed, accuracy, run-up speeds), physical demands (global positioning system, counter-movement jump), physiological (heart rate, core temperature, skin temperature, sweat loss), biochemical (creatine kinase, C-reactive protein) and perceptual variables (perceived exertion, thermal sensation, muscle soreness). Mean ball speed was higher after cooling in Session 2 (118.9 ± 8.1 vs. 115.5 ± 8.6 km · h−1; P = 0.001; d = 0.67), reducing declines in ball speed between sessions (0.24 vs. −3.18 km · h−1; P = 0.03; d = 1.80). Large effects indicated higher accuracy in Session 2 after cooling (46.0 ± 11.2 vs. 39.4 ± 8.6 arbitrary units [AU]; P = 0.13; d = 0.93) without affecting total run-up speed (19.0 ± 3.1 vs. 19.0 ± 2.5 km · h−1; P = 0.97; d = 0.01). Cooling reduced core temperature, skin temperature and thermal sensation throughout the intervention (P = 0.001–0.05; d = 1.31–5.78) and attenuated creatine kinase (P = 0.04; d = 0.56) and muscle soreness at 24-h (P = 0.03; d = 2.05). Accordingly, mixed-method cooling can reduce thermal strain after a 10-over spell and improve markers of muscular damage and discomfort alongside maintained medium-fast bowling performance on consecutive days in hot conditions.

Comparative effects of posterior eye cup tissues from Myopic and Hyperopic chick eyes on cultured scleral fibroblasts

The role of individual ocular tissues in mediating changes to the sclera during myopia development is unclear. The aim of this study was to examine the effects of retina, RPE and choroidal tissues from myopic and hyperopic chick eyes on the DNA and glycosaminoglycan (GAG) content in cultures of chick scleral fibroblasts. Primary cultures of fibroblastic cells expressing vimentin and -smooth muscle actin were established in serum-supplemented growth medium from 8-day-old normal chick sclera. The fibroblasts were subsequently co-cultured with posterior eye cup tissue (full thickness containing retina, RPE and choroid) obtained from untreated eyes and eyes wearing translucent diffusers (form-deprivation myopia, FDM) or -15D lenses (lens-induced myopia, LIM) for 3 days (post hatch day 5 to 8) (n=6 per treatment group). The effect of tissues (full thickness and individual retina, RPE, and choroid layers) from -15D (LIM) versus +15D (lens-induced hyperopia, LIH) treated eyes was also determined. Refraction changes in the direction predicted by the visual treatments were confirmed by retinoscopy prior to tissue collection. Glycosaminoglycan (GAG) and DNA content of the scleral fibroblast cultures were measured using GAG and PicoGreen assays. There was no significant difference in the effect of full thickness tissue from either FDM or LIM treated eyes on DNA and GAG content of scleral fibroblasts (DNA 8.9±2.6 µg and 8.4±1.1 µg, p=0.12; GAG 11.2±0.6 µg and 10.1±1.0 µg, p=0.34). Retina from LIM eyes did not alter fibroblast DNA or GAG content compared to retina from LIH eyes (DNA 27.2±1.7 µg versus 23.2±1.5 µg, p=0.21; GAG 28.1±1.7 µg versus. 28.7±1.2 µg, p=0.46). Similarly, the choroid from LIH and LIM eyes did not produce a differential effect on DNA content (DNA, LIM 46.9±6.4 versus LIH 51.5±4.7 µg, p=0.31), whereas GAG content was higher for cells in co-culture with choroid from LIH eyes (GAG 32.5±0.7 µg versus 18.9±1.2 µg, F1,6=9.210, p=0.0002). In contrast, fibroblast DNA was greater in co-culture with RPE from LIM eyes than the empty basket and DNA content less for co-culture with RPE from LIH eyes (LIM: 72.4±6.3 µg versus Empty basket: 46.03±1.0 µg; F1,6=69.99, p=0.0005 and LIH: 27.9±2.3 µg versus empty basket: 46.03±1.0 µg; p=0.0004). GAG content was higher with RPE from LIH eyes (LIH: 33.7±1.9 µg versus empty basket: 29.5±0.8 µg, F1,6=13.99, p=0.010) and lower with RPE from LIM eyes (LIM: 27.7±0.9 µg versus empty basket: 29.5±0.8 µg, p=0.021). GAG content of cells in co-culture with choroid from LIH eyes was higher compared to co-culture with choroid from LIM eyes (32.5±0.7 µg versus 18.9±1.2 µg respectively, F1,6=9.210, p=0.0002). In conclusion, these experiments provide evidence for a directional growth signal that is present (and remains) in the ex-vivo RPE, but that does not remain in the ex-vivo retina. The identity of this factor(s) that can modify scleral cell DNA and GAG content requires further research.

A coupled SPH-DEM model for fluid and solid mechanics of apple parenchyma cells during drying

A coupled SPH-DEM based two-dimensional (2-D) micro-scale single cell model is developed to predict basic cell-level shrinkage effects of apple parenchyma cells during air drying. In this newly developed drying model, Smoothed Particle Hydrodynamics (SPH) is used to model the low Reynolds Number fluid motions of the cell protoplasm, and a Discrete Element Method (DEM) is employed to simulate the polymer-like cell wall. Simulations results reasonably agree with published experimental drying results on cellular shrinkage properties such as cellular area, diameter and perimeter. These preliminary results indicate that the model is effective for the modelling and simulation of apple parenchyma cells during air drying.

Effects of various power system stabilizers on improving power system dynamic performance

To ensure the small-signal stability of a power system, power system stabilizers (PSSs) are extensively applied for damping low frequency power oscillations through modulating the excitation supplied to synchronous machines, and increasing interest has been focused on developing different PSS schemes to tackle the threat of damping oscillations to power system stability. This paper examines four different PSS models and investigates their performances on damping power system dynamics using both small-signal eigenvalue analysis and large-signal dynamic simulations. The four kinds of PSSs examined include the Conventional PSS (CPSS), Single Neuron based PSS (SNPSS), Adaptive PSS (APSS) and Multi-band PSS (MBPSS). A steep descent parameter optimization algorithm is employed to seek the optimal PSS design parameters. To evaluate the effects of these PSSs on improving power system dynamic behaviors, case studies are carried out on an 8-unit 24-bus power system through both small-signal eigenvalue analysis and large-signal time-domain simulations.

The effects of truth in the hinterlands and abused [in Portugese] Os efeitos de verdade em os sertões e abusado

O presente livro propõe-se a refletir sobre a construção dos efeitos de verdade em dois modos de produção de ordem discursiva: Os sertões, romance de Euclides da Cunha, e Abusado: o dono do morro Dona Marta, romance-reportagem do jornalista Caco Barcellos. A autora apresenta a fragilidade do discurso jornalístico em legitimar-se como o discurso do verdadeiro, em dura oposição ao discurso da literatura, lugar da ficção, expressão dos sentimentos, imaginário, da não-verdade. [Portugese]

Effects of acute multi-nutrient supplementation on rugby union match performance and recovery

Aim: To determine the effects of an acute multi-nutrient supplement on physiological, performance and recovery responses to intermittent-sprint running and muscular damage during rugby union matches. Methods: Using a randomised, double-blind, cross-over design, twelve male rugby union players ingested either 75 g of a comprehensive multi-nutrient supplement (SUPP), [Musashi] or 1 g of a taste and carbohydrate matched placebo (PL) for 5 days pre-competition. Competitive rugby union game running performance was then measured using 1 Hz GPS data (SPI10, SPI elite, GPSports), in addition to associated blood draws, vertical jump assessments and ratings of perceived muscular soreness (MS) pre, immediately post and 24 h post-competition. Baseline (BL) GPS data was collected during six competition rounds preceding data collection. Results: No significant differences were observed between supplement conditions for all game running, vertical jump, and ratings of perceived muscular soreness. However, effect size analysis indicated SUPP ingestion increased 1st half very high intensity running (VHIR) mean speed (d = 0.93) and 2nd half relative distance (m/min) (d = 0.97). Further, moderate increases in 2nd half VHIR distance (d = 0.73), VHIR m/min (d = 0.70) and VHIR mean speed (d = 0.56) in SUPP condition were also apparent. Moreover, SUPP demonstrated significant increases in 2nd half dist m/min, total game dist m/min and total game HIR m/min compared with BL data (P < 0.05). Further, large ES increases in VHIR time (d = 0.88) and moderate increases in 2nd half HIR m/min (d = 0.65) and 2nd half VHIR m/min (d = 0.74) were observed between SUPP and BL. Post-game aspartate aminotransferase (AST) (d = 1.16) and creatine kinase (CK) (d = 0.97) measures demonstrated increased ES values with SUPP, while AST and CK values correlated with 2nd half VHIR distance (r = −0.71 and r = −0.76 respectively). Elevated c-reactive protein (CRP) was observed post-game in both conditions, however was significantly blunted with SUPP (P = 0.05). Additionally, pre-game (d = 0.98) and post-game (d = 0.96) increases in cortisol (CORT) were apparent with SUPP. No differences were apparent between conditions for pH, lactate, glucose, HCO3, vertical jump assessments and MS (P > 0.05). Conclusion: These findings suggest SUPP may assist in the maintenance of VHIR speeds and distances covered during rugby union games, possibly via the buffering qualities of SUPP ingredients (i.e. caffeine, creatine, bicarbonate). While the mechanisms for these findings are unclear, the similar pH between conditions despite additional VHIR during SUPP may support this conclusion. Finally, correlations between increased work completed at very high intensities and muscular degradation in SUPP conditions, may mask any anti-catabolic properties of supplementation.

Effects of oxygen and culture system on in vitro propagation and redifferentiation of osteoarthritic human articular chondrocytes

Regenerative medicine-based approaches for the repair of damaged cartilage rely on the ability to propagate cells while promoting their chondrogenic potential. Thus, conditions for cell expansion should be optimized through careful environmental control. Appropriate oxygen tension and cell expansion substrates and controllable bioreactor systems are probably critical for expansion and subsequent tissue formation during chondrogenic differentiation. We therefore evaluated the effects of oxygen and microcarrier culture on the expansion and subsequent differentiation of human osteoarthritic chondrocytes. Freshly isolated chondrocytes were expanded on tissue culture plastic or CultiSpher-G microcarriers under hypoxic or normoxic conditions (5% or 20% oxygen partial pressure, respectively) followed by cell phenotype analysis with flow cytometry. Cells were redifferentiated in micromass pellet cultures over 4 weeks, under either hypoxia or normoxia. Chondrocytes cultured on tissue culture plastic proliferated faster, expressed higher levels of cell surface markers CD44 and CD105 and demonstrated stronger staining for proteoglycans and collagen type II in pellet cultures compared with microcarrier-cultivated cells. Pellet wet weight, glycosaminoglycan content and expression of chondrogenic genes were significantly increased in cells differentiated under hypoxia. Hypoxia-inducible factor-3alpha mRNA was up-regulated in these cultures in response to low oxygen tension. These data confirm the beneficial influence of reduced oxygen on ex vivo chondrogenesis. However, hypoxia during cell expansion and microcarrier bioreactor culture does not enhance intrinsic chondrogenic potential. Further improvements in cell culture conditions are therefore required before chondrocytes from osteoarthritic and aged patients can become a useful cell source for cartilage regeneration.

Molecular dynamics studies of the effects of branching characteristics on the crystalline structure of polyethylene

Molecular dynamics simulations were carried out on single chain models of linear low-density polyethylene in vacuum to study the effects of branch length, branch content, and branch distribution on the polymer’s crystalline structure at 300 K. The trans/gauche (t/g) ratios of the backbones of the modeled molecules were calculated and utilized to characterize their degree of crystallinity. The results show that the t/g ratio decreases with increasing branch content regardless of branch length and branch distribution, indicating that branch content is the key molecular parameter that controls the degree of crystallinity. Although t/g ratios of the models with the same branch content vary, they are of secondary importance. However, our data suggests that branch distribution (regular or random) has a significant effect on the degree of crystallinity for models containing 10 hexyl branches/1,000 backbone carbons. The fractions of branches that resided in the equilibrium crystalline structures of the models were also calculated. On average, 9.8% and 2.5% of the branches were found in the crystallites of the molecules with ethyl and hexyl branches while C13 NMR experiments showed that the respective probabilities of branch inclusion for ethyl and hexyl branches are 10% and 6% [Hosoda et al., Polymer 1990, 31, 1999–2005]. However, the degree of branch inclusion seems to be insensitive to the branch content and branch distribution.

Executive function development and stress effects on driving performance : preliminary findings from a young adult sample.

This chapter presents a pilot study examining the interactive contributions of executive function development/impairment and psychosocial stress to young adults’ (17-30 years old) driving behaviour in a simulator city scenario.