Chimaeric HIV-1 subtype C Gag molecules with large in-frame C-terminal polypeptide fusions form virus-like particles

HIV-1 Pr55 Gag virus-like particles (VLPs) are strong immunogens with potential as candidate HIV vaccines. VLP immunogenicity can be broadened by making chimaeric Gag molecules: however, VLPs incorporating polypeptides longer than 200 aa fused in frame with Gag have not yet been reported. We constructed a range of gag-derived genes encoding in-frame C-terminal fusions of myristoylation-competent native Pr55Gag and p6-truncated Gag (Pr50Gag) to test the effects of polypeptide length and sequence on VLP formation and morphology, in an insect cell expression system. Fused sequences included a modified reverse transcriptase-Tat-Nef fusion polypeptide (RTTN, 778 aa), and truncated versions of RTTN ranging from 113 aa to 450 aa. Baculovirus-expressed chimaeric proteins were examined by western blot and electron microscopy. All chimaeras formed VLPs which could be purified by sucrose gradient centrifugation. VLP diameter increased with protein MW, from ∼100 nm for Pr55Gag to ∼250 nm for GagRTTN. The presence or absence of the Gag p6 region did not obviously affect VLP formation or appearance. GagRT chimaeric particles were successfully used in mice to boost T-cell responses to Gag and RT that were elicited by a DNA vaccine encoding a GagRTTN polypeptide, indicating the potential of such chimaeras to be used as candidate HIV vaccines. © 2008 Elsevier B.V. All rights reserved.

A prime-boost immunisation regimen using recombinant BCG and Pr55 gag virus-like particle vaccines based on HIV type 1 subtype C successfully elicits Gag-specific responses in baboons

Mycobacterium bovis BCG is considered an attractive live bacterial vaccine vector. In this study, we investigated the immune response of baboons to a primary vaccination with recombinant BCG (rBCG) constructs expressing the gag gene from a South African HIV-1 subtype C isolate, and a boost with HIV-1 subtype C Pr55 gag virus-like particles (Gag VLPs). Using an interferon enzyme-linked immunospot assay, we show that although these rBCG induced only a weak or an undetectable HIV-1 Gag-specific response on their own, they efficiently primed for a Gag VLP boost, which strengthened and broadened the immune responses. These responses were predominantly CD8+ T cell-mediated and recognised similar epitopes as those targeted by humans with early HIV-1 subtype C infection. In addition, a Gag-specific humoral response was elicited. These data support the development of HIV-1 vaccines based on rBCG and Pr55 gag VLPs. © 2009 Elsevier Ltd. All rights reserved.

HIV-1 subtype C Pr55gag virus-like particle vaccine efficiently boosts baboons primed with a matched DNA vaccine

A DNA vaccine expressing human immunodeficiency virus type 1 (HIV-1) southern African subtype C Gag (pTHGag) and a recombinant baculovirus Pr55gag virus-like particle prepared using a subtype C Pr55gag protein (Gag VLP) was tested in a prime-boost inoculation regimen in Chacma baboons. The response of five baboons to Gag peptides in a gamma interferon (IFN-γ) enzyme-linked immunospot (ELISPOT) assay after three pTHGag immunizations ranged from 100 to 515 spot-forming units (s.f.u.) per 106 peripheral blood mononuclear cells (PBMCs), whilst the response of two baboons to the Gag VLP vaccine ranged from 415 to 465 s.f.u. per 106 PBMCs. An increase in the Gag-specific response to a range of 775-3583 s.f.u. per 106 PBMCs was achieved by boosting with Gag VLPs the five baboons that were primed with pTHGag. No improvement in Gag responses was achieved in this prime-boost inoculation regimen by increasing the number of pTHGag inoculations to six. IFN-γ responses were mapped to several peptides, some of which have been reported to be targeted by PBMCs from HIV-1 subtype C-infected individuals. Gag VLPs, given as a single-modality regimen, induced a predominantly CD8+ T-cell IFN-γ response and interleukin-2 was a major cytokine within a mix of predominantly Th1 cytokines produced by a DNA-VLP prime-boost modality. The prime-boost inoculation regimen induced high serum p24 antibody titres in all baboons, which were several fold above that induced by the individual vaccines. Overall, this study demonstrated that these DNA prime/VLP boost vaccine regimens are highly immunogenic in baboons, inducing high-magnitude and broad multifunctional responses, providing support for the development of these products for clinical trials. © 2008 SGM.

An investigation into the use of human papillomavirus type 16 virus-like particles as a delivery vector system for foreign proteins: N- and C-terminal fusion of GFP to the L1 and L2 capsid proteins

Development of vaccine strategies against human papillomavirus (HPV), which causes cervical cancer, is a priority. We investigated the use of virus-like particles (VLPs) of the most prevalent type, HPV-16, as carriers of foreign proteins. Green fluorescent protein (GFP) was fused to the N or C terminus of both L1 and L2, with L2 chimeras being co-expressed with native L1. Purified chimaeric VLPs were comparable in size (∼55 nm) to native HPV VLPs. Conformation-specific monoclonal antibodies (Mabs) bound to the VLPs, thereby indicating that they possibly retain their antigenicity. In addition, all of the VLPs encapsidated DNA in the range of 6-8 kb. © 2007 Springer-Verlag.

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.

DRD2/ANKK1 Taq1A (rs 1800497 C>T) genotypes are associated with susceptibility to second generation antipsychotic-induced akathisia

Rationale Although the advent of atypical, second-generation antipsychotics (SGAs) has resulted in reduced likelihood of akathisia, this adverse effect remains a problem. Extrapyramidal adverse effects are associated with increased drug occupancy of dopamine 2 receptors (DRD2). The A1 allele of the DRD2/ANKK1,rs1800497, is associated with decreased striatal DRD2 density. Objectives The aim of this study was to identify whether the A1(T) allele of the DRD2/ANKK1 was associated with akathisia (measured with the Barnes Akathisia Rating Scale) in a clinical sample of 234 patients treated with antipsychotics. Results Definite akathisia (a score≥ 2 for the global clinical assessment of akathisia) was significantly less common in subjects prescribed SGAs (16.8 %) than those prescribed FGAs (47.6%), p<0.0001. Overall, 24.1% of A1+ (A1A2/A1A1) patients treated with SGAs had akathisia compared to 10.8% of A1- (A2A2) patients. A1+ (A1A2/A1A1) patients administered SGAs also had higher global clinical assessment of akathisia scores than A1- subjects (p=0.01). SGAs maintained their advantage over FGAs regarding akathisia even in A1+ patients treated with SGAs. Conclusions These results strongly suggest that A1+ variants of the DRD2/ANKK1 Taq1A allele confer risk for akathisia in patients treated with SGAs and may explain inconsistencies across prior studies comparing FGAs and SGAs.

The effect of post-match alcohol ingestion on recovery from competitive Rugby League matches

This study investigated the effects of alcohol ingestion on lower body strength and power, and physiological and cognitive recovery following competitive Rugby League matches. Nine male Rugby players participated in two matches, followed by one of two randomized interventions; a control or alcohol ingestion session. Four hours post-match, participants consumed either beverages containing a total of 1g of ethanol per kg bodyweight (vodka and orange juice; ALC) or a caloric and taste matched non-alcoholic beverage (orange juice; CONT). Pre, post, 2 h post and 16 h post match measures of countermovement jump (CMJ), maximal voluntary contraction(MVC), voluntary activation (VA), damage and stress markers of creatine kinase (CK), C-reactive protein (CRP), cortisol, and testosterone analysed from venous blood collection, and cognitive function (modified Stroop test) were determined. Alcohol resulted in large effects for decreased CMJ height(-2.35 ± 8.14 and -10.53 ± 8.36 % decrement for CONT and ALC respectively; P=0.15, d=1.40), without changes in MVC (P=0.52, d=0.70) or VA (P=0.15, d=0.69). Furthermore, alcohol resulted in a significant slowing of total time in a cognitive test (P=0.04, d=1.59), whilst exhibiting large effects for detriments in congruent reaction time (P=0.19, d=1.73). Despite large effects for increased cortisol following alcohol ingestion during recovery (P=0.28, d=1.44), post-match alcohol consumption did not unduly affect testosterone (P-0.96, d=0.10), CK (P=0.66, d=0.70) or CRP(P=0.75, d=0.60). It appears alcohol consumption during the evening following competitive rugby matches may have some detrimental effects on peak power and cognitive recovery the morning following a Rugby League match. Accordingly, practitioners should be aware of the potential associated detrimental effects of alcohol consumption on recovery and provide alcohol awareness to athletes at post-match functions.

Near work-induced contrast adaptation in emmetropic and myopic children

Purpose. Contrast adaptation may induce an error signal for emmetropization. This research aims to determine whether reading causes contrast adaptation in children and, if so, to determine whether myopes exhibit greater contrast adaptation than emmetropes. Methods. Baseline contrast sensitivity was determined in 34 emmetropic and 34 spectacle-corrected myopic children for 0.5, 1.2, 2.7, 4.4, and 6.2 cycles per degree (cpd) horizontal sine-wave gratings. Effects of near tasks on contrast sensitivity were determined during periods spent looking at a 6.2 cpd horizontal grating and during periods spent reading lines of English text, with 1.2 cpd row frequency and 6 cpd stroke frequency. Results. Both emmetropic and myopic groups (mean ± SD; age, 10.3 ± 1.4 years) showed reduced contrast sensitivity during both near tasks, with greatest overall adaptation at 6.2 cpd. Adaptation induced by viewing the grating (0.15 ± 0.17 log unit [40%]; range, 0.07-0.27 log unit) was significantly greater than adaptation induced by reading text (0.11 ± 0.18 log unit [29%], 0.08-0.16 log unit) (F(1,594) = 10.7; P = 0.001). Myopic children showed significantly greater adaptation across the tasks (0.15 ± 0.18 log unit [42%]) than emmetropic children (0.10 ± 0.16 log unit [26%]) (F(1,66) = 7.30; P = 0.009), with the greatest difference occurring at 4.4 cpd (mean, 0.11 log unit [30%]). Conclusions. Grating and reading tasks induced contrast adaptation; viewing horizontal gratings induced greater adaptation than reading, and myopes exhibited greater adaptation than emmetropes. Contrast adaptation effects may underlie findings of prolonged near work being associated with myopia. However, our research does not show whether this is consequential or causal.

Prevalence of malnutrition in community-dwelling adults with Parkinson's disease

People with Parkinson’s disease (PD) have been reported to be at higher risk of malnutrition than an age-matched population due to PD motor and non-motor symptoms and pharmacotherapy side effects. The prevalence of malnutrition in PD has yet to be well-defined. Community-dwelling people with PD, aged > 18 years, were recruited (n = 97, 61 M, 36 F). The Patient-Generated Subjective Global Assessment (PGSGA) was used to assess nutritional status, the Parkinson’s Disease Questionnaire (PDQ-39) was used to assess quality of life, and the Beck’s Depression Inventory (BDI) was used to measure depression. Levodopa equivalent doses (LEDs) were calculated based on reported Parkinson’s disease medication. Weight, height, mid-arm circumference (MAC) and calf circumference were measured. Cognitive function was measured using the Addenbrooke’s Cognitive Examination. Average age was 70.0 (9.1, 35–92) years. Based on SGA, 16 (16.5%) were moderately malnourished (SGA B) while none were severely malnourished (SGA C). The well-nourished participants (SGA A) had a better quality of life, t(90) = −2.28, p < 0.05, and reported less depressive symptoms, t(94)= −2.68, p < 0.05 than malnourished participants. Age, years since diagnosis, cognitive function and LEDs did not signifi cantly differ between the groups. The well-nourished participants had lower PG-SGA scores, t(95) = −5.66, p = 0.00, higher BMIs, t(95) = 3.44, p < 0.05, larger MACs, t(95) = 3.54, p < 0.05 and larger calf circumferences, t(95) = 2.29, p < 0.05 than malnourished participants. Prevalence of malnutrition in community-dwelling adults with PD in this study is comparable to that in other studies with community-dwelling adults without PD and is higher than other PD studies where a nutritional status assessment tool was used. Further research is required to understand the primary risk factors for malnutrition in this group.

Automated speed enforcement in Australia : recent examples of the influence of public opinion on program sustainability

In Australia, speeding remains a substantial contributor to road trauma. The National Road Safety Strategy (2011-2020) highlighted the need to harness community support for current and future speed management strategies. Australia is known for intensive speed camera programs which are both automated and manual, employing covert and overt methods. Recent developments in the area of automated speed enforcement in Australia help to illustrate the important link between community attitudes to speed enforcement and subsequent speed camera policy developments. A perceived lack of community confidence in camera programs prompted reviews in New South Wales and Victoria in 2011 by the jurisdictional Auditor-General. This paper explores automated speed camera enforcement in Australia with particular reference to the findings of these two reports as they relate to the level of public support for and community attitudes towards automated speed enforcement. It also provides comment on the evolving nature of automated speed enforcement according to previously identified controversies and dilemmas associated with speed camera programs.

Effect of bone morphogenetic protein-4 (BMP-4) on the expression of Sox2, Oct-4 and c-Myc in human periodontal ligament cells during long-term culture

Recent studies demonstrated endogenous expression level of Sox2, Oct-4 and c-Myc is correlated with the pluripotency and successful induction of induced pluripotent stem cells (iPSCs). Periondontal ligament cells (PDLCs)have multi-lineage diferentiation capability and ability to maintain undifferentiated stage, which makes PDLCs a suitable cell source for tissue repair and regeneration. To elucidate the effect of in vitro culture condition on the stemness potential of PDLCs, we explored the cell growth, proliferation, cell cycle, and the expression of Sox2, Oct-4 and c-Myc in PDLCs from passage 1 to 7 with or without the addition of recombinant human BMP4(rhBMP4). Our results revealed that BMP-4 promoted cell growth and proliferation, arrested PDLCs in S phase of cell cycle and upregulated PI value. It was revealed that without the addition of rhBMP4, the expression of Sox2, Oct-4 and c-Myc in PDLCs only maintained nucleus location until passage 3, then lost nucleus location subsequently. The mRNA expression in PDLCs further confirmed that the level of Sox2 and Oct-4 peaked at passage 3, then decreased afterwards, whereas c-Myc maintained consistently upregulation along passages. after the treatment with rhBMP4, the expression of Sox2, Oct-4 and c-Myc in PDLCs maintained nucleus location even at passage 7 and the mRNA expression of Sox2 and Oct-4 significantly upregulated at passage 5 and 7. These results demonstrated that addition of rhBMP-4 in the culture media could improve the current culture condition for PDLCs to maintain in an undifferentiated stage.

RcppArmadillo : accelerating R with high-performance C++ linear algebra

The R statistical environment and language has demonstrated particular strengths for interactive development of statistical algorithms, as well as data modelling and visualisation. Its current implementation has an interpreter at its core which may result in a performance penalty in comparison to directly executing user algorithms in the native machine code of the host CPU. In contrast, the C++ language has no built-in visualisation capabilities, handling of linear algebra or even basic statistical algorithms; however, user programs are converted to high-performance machine code, ahead of execution. A new method avoids possible speed penalties in R by using the Rcpp extension package in conjunction with the Armadillo C++ matrix library. In addition to the inherent performance advantages of compiled code, Armadillo provides an easy-to-use template-based meta-programming framework, allowing the automatic pooling of several linear algebra operations into one, which in turn can lead to further speedups. With the aid of Rcpp and Armadillo, conversion of linear algebra centered algorithms from R to C++ becomes straightforward. The algorithms retains the overall structure as well as readability, all while maintaining a bidirectional link with the host R environment. Empirical timing comparisons of R and C++ implementations of a Kalman filtering algorithm indicate a speedup of several orders of magnitude.

Development of a survey instrument to measure TEFL academics' perceptions about, individual and workplace characteristics for conducting research

The Chinese Ministry of Education (MOE) initiatives to improve the English competence of college students, as well as increased proficiency level of entering college students (Cheng, 2002) have contributed to greater demands on Teaching English as a Foreign Language (TEFL) academics (MOE, 2004), as “the upgrading of national English proficiency, then, is predicated largely on the professional competence of the teaching force” (Hu, 2005, p. 655). For TEFL academics, one component of this competence is the capacity to conduct research (Shu, 2002), which also reflects other changes in Chinese higher education. The aspirations of higher education institutions at all levels have led to more rigorous recruitment policies and promotion requirements (Che, 2004; Wang, 2007), stressing research as an important indicator of academics’ performance (Shi, 2002; Pan, 2006). These changes highlight the role of research in higher education institutions’ efforts to raise their national status and world ranking (Zhang, Wang, & He, 2006), and have exerted influences on faculty’s academic role. Academics are obliged to engage in research activities, and this has posed challenges to teaching-oriented institutions and disciplines.

C-BN Single-Walled Nanotubes from Hybrid Connection of BN/C Nanoribbons: Prediction by ab initio Density Functional Calculations

We demonstrated for the first time by ab initio density functional calculation and molecular dynamics simulation that C0.5(BN)0.5 armchair single-walled nanotubes (NT) are gapless semiconductors and can be spontaneously formed via the hybrid connection of graphene/BN Nanoribbons (GNR/BNNR) at room temperature. The direct synthesis of armchair C0.5(BN)0.5 via the hybrid connection of GNR/BNNR is predicted to be both thermodynamically and dynamically stable. Such novel armchair C0.5(BN)0.5 NTs possess enhanced conductance as that observed in GNRs. Additionally, the zigzag C0.5(BN)0.5 SWNTs are narrow band gap semiconductors, which may have potential application for light emission. In light of recent experimental progress and the enhanced degree of control in the synthesis of GNRs and BNNR, our results highlight an interesting avenue for synthesizing a novel specific type of C0.5(BN)0.5 nanotube (gapless or narrow direct gap semiconductor), with potentially important applications in BNC-based nanodevices.

Breast cancer detection from thermal images using bispectral invariant features

Highly sensitive infrared (IR) cameras provide high-resolution diagnostic images of the temperature and vascular changes of breasts. These images can be processed to emphasize hot spots that exhibit early and subtle changes owing to pathology. The resulting images show clusters that appear random in shape and spatial distribution but carry class dependent information in shape and texture. Automated pattern recognition techniques are challenged because of changes in location, size and orientation of these clusters. Higher order spectral invariant features provide robustness to such transformations and are suited for texture and shape dependent information extraction from noisy images. In this work, the effectiveness of bispectral invariant features in diagnostic classification of breast thermal images into malignant, benign and normal classes is evaluated and a phase-only variant of these features is proposed. High resolution IR images of breasts, captured with measuring accuracy of ±0.4% (full scale) and temperature resolution of 0.1 °C black body, depicting malignant, benign and normal pathologies are used in this study. Breast images are registered using their lower boundaries, automatically extracted using landmark points whose locations are learned during training. Boundaries are extracted using Canny edge detection and elimination of inner edges. Breast images are then segmented using fuzzy c-means clustering and the hottest regions are selected for feature extraction. Bispectral invariant features are extracted from Radon projections of these images. An Adaboost classifier is used to select and fuse the best features during training and then classify unseen test images into malignant, benign and normal classes. A data set comprising 9 malignant, 12 benign and 11 normal cases is used for evaluation of performance. Malignant cases are detected with 95% accuracy. A variant of the features using the normalized bispectrum, which discards all magnitude information, is shown to perform better for classification between benign and normal cases, with 83% accuracy compared to 66% for the original.