Effects of modifying the tRNA(3Lys) anticodon on the initiation of human immunodeficiency virus type 1 reverse transcription

tRNA(3Lys) is a primer for reverse transcription in human immunodeficiency virus type 1 (HIV-1), and the anticodon of tRNA(3Lys) has been implicated in playing a role in both its placement onto the HIV-1 genome and its interaction with HIV-1 reverse transcriptase (RT). In this work, the anticodon in a tRNA(3Lys) gene was changed from UUU to CUA (tRNA(3Lys)Su+) or, in addition, G-73 was altered to A (tRNA(3Lys)Su+G73A). COS-7 cells were transfected with either wild-type or mutant tRNA(3Lys) genes, and both the wild-type and mutant tRNA(3Lys) produced were purified by using immobilized tRNA-specific hybridization probes. Each mutant tRNA(3Lys) was tested for its ability to prime reverse transcription in vitro, either alone or in competition with wild-type tRNA(3Lys). Short RT extensions of wild-type and mutant tRNALys could be distinguished from each other by their different mobilities in one-dimensional single-stranded conformation polymorphism polyacrylamide gel electrophoresis. These reverse transcription products show that heat-annealed tRNA(3Lys)Su+ has the same ability as heat-annealed wild-type tRNA(3Lys) to prime RT and competes equally well with wild-type tRNA(3Lys) for priming RT. tRNA(3Lys)Su+G73A has 60% of the wild-type ability to prime RT but competes poorly with wild-type tRNA(3Lys) for priming RT. However, the priming abilities of wild-type and mutant tRNA(3) are quite different when in vivo-placed tRNA is examined. HIV-1 produced in COS cells transfected with a plasmid containing both the HIV-1 proviral DNA and DNA coding for tRNA(3Lys)Su+ contains both endogenous, cellular wild-type tRNA(3Lys) and mutant tRNA(3Lys). When total viral RNA is used as the source of primer tRNA placed onto the genomic RNA in vivo, only wild-type tRNA(3Lys) is used as a primer. If the total viral RNA is first heated and exposed to hybridizing conditions, then both the wild-type and mutant tRNA(3Lys) act as primers for RT. These results indicate that the tRNA(3Lys)Su+ packaged into the virions is unable to act as a primer for RT, and a model is proposed to explain the disparate results between heat-annealed and in vivo-placed primer tRNA.

Using small reverse cycle air conditioners in relocatable classrooms - a case study

A 9-month study of four relocatable school buildings, each retro-fitted with small reverse cycle air conditioners (ACs), was conducted to investigate their effectiveness in heating and cooling the classrooms. A comparison with data from previous studies found the energy used by the ACs for heating these temporary classrooms was only 19–20% of the energy used by individual gas heaters installed in permanent classrooms. When equipment efficiencies were considered, the AC units supplied 20–27% less energy to heat the classrooms. The possible reasons for this reduction in supplied energy are explored in this paper. CO2 emissions for the AC units in heating mode, however, were calculated to be 16% greater than for individual gas heaters. The AC units were also used for cooling and on an average the total annual energy consumption for heating and cooling was found to be 11.6 kWh m−2. Responses to a small survey of staff and students about the use and operation of the conditioners are presented. Their responses were more favourable than the predictions of comfort levels in the classrooms using the Predicted Mean Vote–Predicted Percentage of Dissatisfaction (PMV–PPD) model, which indicated “uncomfortable” conditions on average summer days at 3:00 p.m. and average winter days at 10:00 a.m. Background noise levels in the classrooms with the air conditioners in use were above the recommended maximum design level of 45 dB(A); levels of up to 65 dB(A) were measured.

Lupin kernel fiber consumption modifies fecal microbiota in healthy men as determined by rRNA gene flourescent in situ by hybridization

Background Changes in the composition of gastrointestinal microbiota by dietary interventions using pro- and prebiotics provide opportunity for improving health and preventing disease. However, the capacity of lupin kernel fiber (LKFibre), a novel legume-derived food ingredient, to act as a prebiotic and modulate the colonic microbiota in humans needed investigation.

Aim of the study The present study aimed to determine the effect of LKFibre on human intestinal microbiota by quantitative fluorescent in situ hybridization (FISH) analysis.

Design A total of 18 free-living healthy males between the ages of 24 and 64 years consumed a control diet and a LKFibre diet (containing an additional 17–30 g/day fiber beyond that of the control—incorporated into daily food items) for 28 days with a 28-day washout period in a single-blind, randomized, crossover dietary intervention design.
Methods Fecal samples were collected for 3 days towards the end of each diet and microbial populations analyzed by FISH analysis using 16S rRNA gene-based oligonucleotide probes targeting total and predominant microbial populations.

Results Significantly higher levels of Bifidobacterium spp. (P = 0.001) and significantly lower levels of the clostridia group of C. ramosum, C. spiroforme and C. cocleatum (P = 0.039) were observed on the LKFibre diet compared with the control. No significant differences between the LKFibre and the control diet were observed for total bacteria, Lactobacillus spp., the Eubacterium spp., the C. histolyticum/C. lituseburense group and the Bacteroides–Prevotella group.
Conclusions Ingestion of LKFibre stimulated colonic bifidobacteria growth, which suggests that this dietary fiber may be considered as a prebiotic and may beneficially contribute to colon health.

Effective and strategic use of electronic reverse auctions

This paper reports the findings of an empirical study of Electronic Reverse Auctions (e-RAs). This study seeks to better understand the underlying and emerging issues in e-RAs to comprehend the implications of e-RAs on organizational procurement. The study contributes to practice and research by providing insight into emerging e-RA adoption issues, and by discussing strategies for improving e-RA processes, thus improving the effectiveness of e-RAs.

Reverse osmosis membrane fouling and its control

Reverse osmosis (RO) membranes can be used to treat ground water, surface water and wastewater, but a major problem in RO membrane applications is fouling. Many studies have investigated the mechanisms of fouling in RO membranes and suggested various methods to control the membrane fouling. This paper reviews several aspects of RO membranes, such as the types of membrane fouling, their effects on RO membrane processes and fouling control technologies, focussed on the appropriate feed pretreatment technologies and cleaning methods, especially on the membrane filtration pretreatment processes, including microfiltration (MF), ultrafiltration (UF) and nanofiltration (NF), which are the new trends in designing a pretreatment system in the application of RO.

The case for reverse mortgages in Australia : applying the USA experience

In the USA, reverse mortgages have been promoted as a means of accessing equity locked up in a residence, especially after the owner/s has retired. Although there have been some teething problems, the concept of mortgaging the family home after achieving freehold ownership has many merits. Often an asset-rich household must survive on relatively small regular income, and is unable to access the increasing wealth of the family home. A reverse mortgage overcomes this hurdle.

The largest asset for many ageing households is their primary place of residence, the traditional house and suburban land parcel. Recently, the Australian housing market has witnessed substantial growth in the value of its capital city housing, especially on the east coast of Australia. This can be attributed to factors such as owner-occupiers trading up to a better class of dwelling, and the continuing gentrification process for owners choosing not to relocate. At the same time, demographic changes have placed pressure on the regular income of retirees, many of whom have no superannuation fund. For example, life expectancy rates continue to rise and there are an increasing proportion of single person households in society. This has placed additional pressure on financial resources of retirees, especially those with a substantial investment in their family home and a relatively small pension.

This paper visits the reverse mortgage scenario in the USA and considers potential implications for the Australian market. Strengths and weaknesses of this product are contemplated, and the viability of reverse mortgages is discussed. Although there are obvious benefits for certain segments of society, reverse mortgages are a unique product and caution should be exercised to ensure the public is fully knowledgeable from the outset.

Sperm competition and offspring viability at hybridization in Australian tree frogs, Litoria peronii and L. tyleri

Hybridization between closely related species often leads to reduced viability or fertility of offspring. Complete failure of hybrid offspring (post-zygotic hybrid incompatibilities) may have an important role in maintaining the integrity of reproductive barriers between closely related species. We show elsewhere that in Peron's tree frog, Litoria peronii, males more closely related to a female sire more offspring in sperm competition with a less related rival male. Observations of rare 'phenotypic intermediate' males between L. peronii and the closely related L. tyleri made us suggest that these relatedness effects on siring success may be because of selection arising from risks of costly hybridization between the two species. Here, we test this hypothesis in an extensive sperm competition experiment, which shows that there is no effect of species identity on probability of fertilization in sperm competition trials controlling for sperm concentration and sperm viability. Instead, there was a close agreement between a male's siring success in isolation with a female and his siring success with the same female in competition with a rival male regardless of species identity. Offspring viability and survival, however, were strongly influenced by species identity. Over a 14-day period, hybrid offspring suffered increasing mortality and developed more malformations and an obvious inability to swim and right themselves, leading to compromised probability of survival. Thus, hybridization in these sympatric tree frogs does not compromise fertilization but has a strong impact on offspring viability and opportunity for reinforcement selection on mate choice for conspecific partners.

Reverse ascertainment bias in microsatellite allelic diversity in owls (Aves, Strigiformes)

A rich source of markers may be overlooked by screening for polymorphism in the source species only. We screened 129 microsatellite loci isolated from the powerful owl (Ninox strenua) against two closely related species; Ninox  connivens and Ninox novaeseelandiae. From the screening effort 20 polymorphic markers were isolated, including six loci which were originally discarded as they were monomorphic in the source species. Further cross-species amplification of all 20 loci across species from two families, Strigidae and Tytonidae, revealed unusually high levels of polymorphism within closely related species, and limited success within phylogenetically distant species. Routine screening of multiple  species during the marker development phase can yield a wider range of  polymorphic markers which can subsequently enhance cross-species  amplification attempts.

Reverse microemulsion-mediated synthesis of SiO2-coated ZnO composite nanoparticles : multiple cores with tunable shell thickness

Monodispersed SiO₂-shell/ZnO-core composite nanospheres have been prepared in an oil-in-water microemulsion system. By using cyclohexane as the oil phase and Triton X-100 as the surfactant, composite nanospheres with high core loading levels and tunable shell thickness were obtained. Utilization of PVP capping agent on ZnO allowed the synthesis of composite nanospheres without forming any coreless SiO2 spheres or shell-less ZnO particles. The photoactivity of ZnO nanoparticles was greatly reduced by SiO2-coating, which enables their applications as durable, safe, and nonreactive UV blockers in plastics, coating, and other products.

An analysis of microband orientation in hot deformed aluminium alloy AA5052 using forward and reverse torsion

The effects of strain path reversal on the macroscopic orientation of microbands in AA5052 have been studied using high resolution electron backscatter diffraction. Deformation was carried using two equal steps of forward/forward or forward/reverse torsion at a temperature of 300°C and strain rate of 1s-1 to a total equivalent tensile strain of 0.5. In both cases microbands were found in the majority of grains examined with many having more than one set. The microbands appear to cluster at specific angles to the macroscopic deformation. For the forward/forward condition microbands clustered around -20° and +45° to the maximum principle stress direction and at ± 30-35° to the principal strain direction. For the forward/reverse condition significantly more spread in microband angle was observed though peaks were visible at ±35° with respect to principal stress direction and at -40° and +30° with respect to the principal strain direction of the reverse torsion. This suggests the microbands formed in the forward deformation have or are dissolving and any new microbands formed are related to the deformation conditions of the final strain path.

Diagnosis of {alpha}+-thalassemias by determining the ratio of the two {alpha}-globin gene copies by oligonucleotide hybridization and melting curve analysis

Most cases of {alpha}-thalassemia result from large deletions at the {alpha}-globin locus (1). The {alpha}-globin gene cluster contains a tandem array of 2 nearly identical {alpha}-globin genes (HBA; Fig. 1A ) (2). The {alpha}0-thalassemias are characterized by deletions that inactivate both {alpha}-globin genes of a given chromosome, whereas in {alpha}+-thalassemias, one gene remains functional. The most widespread {alpha}+-thalassemias are those designated –{alpha}3.7 and –{alpha}4.2, according to the lengths of the deleted fragments (3).

Expression of the disintegrin metalloprotease ADAM-10 in prostate cancer and its regulation by dihydrotestosterone, insulin like growth factor -1 and epidermal growth factor in the prostate cell model LNCaP

Purpose: The disintegrin metalloprotease ADAM-10 is a multidomain metalloprotease that is potentially significant in tumor progression due to its extracellular matrix-degrading properties. Previously, ADAM-10 mRNA was detected in prostate cancer (PCa) cell lines; however, the presence of ADAM-10 protein and its cellular localization, regulation, and role have yet to be described. We hypothesized that ADAM-10 mRNA and protein may be regulated by growth factors such as 5α-dihydrotestosterone, insulin-like growth factor I, and epidermal growth factor, known modulators of PCa cell growth and invasion.

Experimental Design: ADAM-10 expression was analyzed by in situ hybridization and immunohistochemistry in prostate tissues obtained from 23 patients with prostate disease. ADAM-10 regulation was assessed using quantitative reverse transcription-PCR and Western blot analysis in the PCa cell line LNCaP.

Results: ADAM-10 expression was localized to the secretory cells of prostate glands, with additional basal cell expression in benign glands. ADAM-10 protein was predominantly membrane bound in benign glands but showed marked nuclear localization in cancer glands. By Western blot, the 100-kDa proform and the 60-kDa active form of ADAM-10 were synergistically up-regulated in LNCaP cells treated with insulin-like growth factor I plus 5α-dihydrotestosterone. Epidermal growth factor also up-regulated both ADAM-10 mRNA and protein.

Conclusions: This study describes for the first time the expression, regulation, and cellular localization of ADAM-10 protein in PCa. The regulation and membrane localization of ADAM-10 support our hypothesis that ADAM-10 has a role in extracellular matrix maintenance and cell invasion, although the potential role of nuclear ADAM-10 is not yet known.

A reverse pathway? Actual and perceived skill proficiency and physical activity

Purpose: Motor skills are considered a prerequisite to physical activity, yet the relationship may be reciprocal and perceived sports competence might mediate associations.
Methods: In 2006/2007, 215 adolescents completed motor skill proficiency (Get Skilled Get Active), perceived sport competence (Physical Self-Perception Profile) and physical activity assessments (Adolescent Physical Activity Recall Questionnaire) as part of the Physical Activity and Skills Study. Using AMOS (Version 7.0), reciprocal relationships were examined between motor skill (object control and locomotor) and moderate to vigorous physical activity (MVPA). Both models were then run in different versions to understand the role of perceived sports competence as a potential mediator.
Results: Mean age was 16.4 yr (SD = 0.6), 51.6% (111/215) were females. A reciprocal relationship between object control and MVPA and a one-way relationship from MVPA to locomotor skill was found. When perceived sports competence was examined as a mediator, the best-fitting model versions explained 16% (R² = 0.16)
MVPA variation, and 30% object control (R² = 0.30), and 12% locomotor skill variation (R² = 0.12) (reverse relationship). Perceived sports competence partially mediates the relationship between object control proficiency and physical activity for both directions and fully mediates the relationship between physical activity and locomotor skill; but only when locomotor skill is the outcome.
Conclusions: If the relationship between object control skill and physical activity is viewed as a ‘‘positive feedback loop,’’ skill development and increasing physical activity should simultaneously be targeted in physical activity interventions. Increasing perceived sport competence should also be an intervention focus.