“We’re not like these weird feather boa-covered AIDS-spreading monsters” : How LGBT young people and service providers think riskiness informs LGBT youth–police interactions

Research has suggested that lesbian, gay, bisexual and transgender (LGBT) young people are “at-risk” of victimization and/or legally “risky.” Relatively few studies have examined the social construction of risk in “risk factor” research and whether risk as a concept influences the everyday lives of LGBT young people. This article reports how 35 LGBT young people and seven service provider staff in Brisbane, Queensland, Australia perceived LGBT youth–police interactions as reflecting discourses about LGBT riskiness and danger. The participants specifically note how they thought looking at-risk and/or looking risky informed their policing experiences. The article concludes with recommendations for improving future policing practice.

"They did not believe me” Responding to child sexual abuse by Church personnel in Australia

Child sexual abuse (CSA) by Church personnel has been subject to study internationally. Such studies have often held a specific focus on the Roman Catholic Church in nations such as Ireland, Belgium and the United States of America (USA). This paper discusses the findings of a study conducted by the author which considers the perspectives of 81 survivors of child sexual abuse by Church personnel in Australia. Participants in this study completed an online survey and then nominated to undertake an in depth interview. The majority of respondents to the survey (66 - 69%) reported experiencing CSA in a Roman Catholic Church, school or children’s home. This paper explores the voices of survivors and recognises the complex and dynamic ways in which they continue to construct and manage their experiences of CSA by Church personnel. In particular, this study considers survivors’ perspectives of the ways in which Churches have responded to their informal disclosures and official complaints of CSA by Church personnel. Similarly to other locations across the world, participants in this study reported feeling re-victimised by Church processes. Participants reported high levels of dissatisfaction with Church policy and procedure in managing child protection issues, as well as high levels of dissatisfaction with the outcome of their complaints.

Not all dimensions of work self-efficacy are equal : understanding the role of tertiary work placements in the development of the elements of work self-efficacy

This paper examines the relationship between a final year tertiary work placement for criminology students at Griffith University in Brisbane and the development of their work self-efficacy. Using a work self-efficacy instrument developed by Professor Joe Raelin at Northeastern University in Boston, a pilot phase in 2006 and a larger study in 2007 investigated the students’ responses across seven self-efficacy factors of learning, problem-solving, teamwork, sensitivity, politics, pressure, and role expectations. Both studies utilised a pre- and post-test and comparisons between these indicated that they believed their abilities to participate constructively in their professional work contexts significantly improved as a result of their placement experience except in the areas of learning, teamwork and sensitivity. This finding will allow us to continue to refine the processes of work placements in order to ensure the integrity of this method for student learning.

Carbonic anhydrase IX expression, a novel surrogate marker of tumor hypoxia, is associated with a poor prognosis in non-small-cell lung cancer

Purpose To evaluate carbonic anhydrase (CA) IX as a surrogate marker of hypoxia and investigate the prognostic significance of different patterns of expression in non-small-cell lung cancer (NSCLC). Methods Standard immunohistochemical techniques were used to study CA IX expression in 175 resected NSCLC tumors. CA IX expression was determined by Western blotting in A549 cell lines grown under normoxic and hypoxic conditions. Measurements from microvessels to CA IX positivity were obtained. Results CA IX immunostaining was detected in 81.8% of patients. Membranous (m) (P = .005), cytoplasmic (c) (P = .018), and stromal (P < .001) CA IX expression correlated with the extent of tumor necrosis (TN). The mean distance from vascular endothelium to the start of tumor cell positivity was 90 μm, which equates to an oxygen pressure of 5.77 mmHg. The distance to blood vessels from individual tumor cells or tumor cell clusters was greater if they expressed mCA IX than if they did not (P < .001). Hypoxic exposure of A549 cells for 16 hours enhanced CAIX expression in the nuclear and cytosolic extracts. Perinuclear (p) CA IX (P = .035) was associated with a poor prognosis. In multivariate analysis, pCA IX (P = .004), stage (P = .001), platelet count (P = .011), sex (P = .027), and TN (P = .035) were independent poor prognostic factors. Conclusion These results add weight to the contention that mCA IX is a marker of tumor cell hypoxia. The absence of CA IX staining close to microvessels suggests that these vessels are functionally active. pCA IX expression is representative of an aggressive phenotype. © 2003 by American Society of Clinical Oncology.

A comparison of the histochemical and image-derived proteoglycan content of articular cartilage

There are several methods for determining the proteoglycan content of cartilage in biomechanics experiments. Many of these include assay-based methods and the histochemistry or spectrophotometry protocol where quantification is biochemically determined. More recently a method based on extracting data to quantify proteoglycan content has emerged using the image processing algorithms, e.g., in ImageJ, to process histological micrographs, with advantages including time saving and low cost. However, it is unknown whether or not this image analysis method produces results that are comparable to those obtained from the biochemical methodology. This paper compares the results of a well-established chemical method to those obtained using image analysis to determine the proteoglycan content of visually normal (n=33) and their progressively degraded counterparts with the protocols. The results reveal a strong linear relationship with a regression coefficient (R2) = 0.9928, leading to the conclusion that the image analysis methodology is a viable alternative to the spectrophotometry.

A study of the phosphate mineral kapundaite NaCa(Fe3+)4(PO4)4(OH)3⋅5(H2O) using SEM/EDX and vibrational spectroscopic methods

Vibrational spectroscopy enables subtle details of the molecular structure of kapundaite to be determined. Single crystals of a pure phase from a Brazilian pegmatite were used. Kapundaite is the Fe3+ member of the wardite group. The infrared and Raman spectroscopy were applied to compare the structure of kapundaite with wardite. The Raman spectrum of kapundaite in the 800–1400 cm−1 spectral range shows two intense bands at 1089 and 1114 cm−1 assigned to the ν1PO43- symmetric stretching vibrations. The observation of two bands provides evidence for the non-equivalence of the phosphate units in the kapundaite structure. The infrared spectrum of kapundaite in the 500–1300 cm−1 shows much greater complexity than the Raman spectrum. Strong infrared bands are found at 966, 1003 and 1036 cm−1 and are attributed to the ν1PO43- symmetric stretching mode and ν3PO43- antisymmetric stretching mode. Raman bands in the ν4 out of plane bending modes of the PO43- unit support the concept of non-equivalent phosphate units in the kapundaite structure. In the 2600–3800 cm−1 spectral range, Raman bands for kapundaite are found at 2905, 3151, 3311, 3449 and 3530 cm−1. These bands are broad and are assigned to OH stretching vibrations. Broad infrared bands are also found at 2904, 3105, 3307, 3453 and 3523 cm−1 and are attributed to water. Raman spectroscopy complimented with infrared spectroscopy has enabled aspects of the structure of kapundaite to be ascertained and compared with that of other phosphate minerals.

Facile mutant identification via a single parental backcross method and application of whole genome sequencing based mapping pipelines

Forward genetic screens have identified numerous genes involved in development and metabolism, and remain a cornerstone of biological research. However, to locate a causal mutation, the practice of crossing to a polymorphic background to generate a mapping population can be problematic if the mutant phenotype is difficult to recognize in the hybrid F2 progeny, or dependent on parental specific traits. Here in a screen for leaf hyponasty mutants, we have performed a single backcross of an Ethane Methyl Sulphonate (EMS) generated hyponastic mutant to its parent. Whole genome deep sequencing of a bulked homozygous F2 population and analysis via the Next Generation EMS mutation mapping pipeline (NGM) unambiguously determined the causal mutation to be a single nucleotide polymorphisim (SNP) residing in HASTY, a previously characterized gene involved in microRNA biogenesis. We have evaluated the feasibility of this backcross approach using three additional SNP mapping pipelines; SHOREmap, the GATK pipeline, and the samtools pipeline. Although there was variance in the identification of EMS SNPs, all returned the same outcome in clearly identifying the causal mutation in HASTY. The simplicity of performing a single parental backcross and genome sequencing a small pool of segregating mutants has great promise for identifying mutations that may be difficult to map using conventional approaches.

Pathogenicity of three RPV isolates of Barley Yellow Dwarf Virus on barley, wheat and wheat alien addition lines

Barleys with and without the Yd2 resistance factor, wheat alien addition stocks with other barley yellow dwarf virus (BYDV) resistance factors and true wheats were challenged with three Australian isolates of BYDV-RPV. Yd2 resistance was effective against two of the BYDV-RPV isolates and inoculated barleys which carry Yd2 did not develop BYD symptoms and shoot growth was not affected. However, barleys with Yd2 were susceptible to the third BYDV-RPV isolate. All barley lines inoculated with the third virus isolate developed typical BYD symptoms (yellowing), shoot growth was reduced compared to uninfected controls and virus titres determined by ELISA were high and similar in barleys with and without Yd2. In contrast, resistances from Thinopyrum intermedium and Agropyron pulcherrimum in wheat backgrounds were effective against all three BYDV-RPV isolates. Shoot growth of inoculated plants with either of these resistance factors did not differ from uninfected controls and virus titres determined by ELISA were very low.

Expression of Caenorhabditis elegans RNA-directed RNA polymerase in transgenic Drosophila melanogaster does not affect morphological development

Drosophila melanogaster, along with all insects and the vertebrates, lacks an RdRp gene. We created transgenic strains of Drosophila melanogaster in which the rrf-1 or ego-1 RdRp genes from C. elegans were placed under the control of the yeast GAL4 upstream activation sequence. Activation of the gene was performed by crossing these lines to flies carrying the GAL4 transgene under the control of various Drosophila enhancers. RT-PCR confirmed the successful expression of each RdRp gene. The resulting phenotypes indicated that introduction of the RdRp genes had no effect on D. melanogaster morphological development. © 2010 Springer Science+Business Media B.V.

Posttranscriptional gene silencing is not compromised in the Arabidopsis CARPEL FACTORY (DICER-LIKE1) mutant, a homolog of dicer-1 from Drosophila

Posttranscriptional silencing (PTGS) in plants, nematodes, Drosophila, and perhaps all eukaryotes operates by sequence-specific degradation or translational inhibition of the target mRNA. These processes are mediated by duplexed RNA. In Drosophila and nematodes, double-stranded (ds)RNA or self-complementary RNA is processed into fragments of approximately 21 nt by Dicer-1 [1, 2]. These small interfering RNAs (siRNAs) serve as guides to target degradation of homologous single-stranded (ss)RNA [1, 3]. In some cases, the approximately 21 nt guide fragments derived from endogenous, imperfectly self-complementary RNAs cause translational inhibition of their target mRNAs, with which they have substantial, but not perfect sequence complementarity [4-6]. These small temporal RNAs (stRNAs) belong to a class of noncoding microRNAs (miRNAs), 20-24 nt in length, that are found in flies, plants, nematodes, and mammals [4, 6-12]. In nematodes, the Dicer-1 enzyme catalyzes the production of both siRNA and stRNA [2, 13-15]. Mutation of the Arabidopsis Dicer-1 homolog, CARPEL FACTORY (CAF), blocks miRNA production [1, 4, 16-18]. Here, we report that the same caf mutant does not block either PTGS or siRNA production induced by self-complementary hairpin RNA. This suggests either that this mutation only impairs miRNA formation or, more interestingly, that plants have two distinct dicer-like enzymes, one for miRNA and another for siRNAi production.

The complete nucleotide sequence of subterranean clover mottle virus

The complete nucleotide sequence of Subterranean clover mottle virus (SCMoV) genomic RNA has been determined. The SCMoV genome is 4,258 nucleotides in length. It shares most nucleotide and amino acid sequence identity with the genome of Lucerne transient streak virus (LTSV). SCMoV RNA encodes four overlapping open reading frames and has a genome organisation similar to that of Cocksfoot mottle virus (CfMV). ORF1 and ORF4 are predicted to encode single proteins. ORF2 is predicted to encode two proteins that are derived from a -1 translational frameshift between two overlapping reading frames (ORF2a and ORF2b). A search of amino acid databases did not find a significant match for ORF1 and the function of this protein remains unclear. ORF2a contains a motif typical of chymotrypsin-like serine proteases and ORF2b has motifs characteristically present in positive-stranded RNA-dependent RNA polymerases. ORF4 is likely to be expressed from a subgenomic RNA and encodes the viral coat protein. The ORF2a/ORF2b overlapping gene expression strategy used by SCMoV and CfMV is similar to that of the poleroviruses and differ from that of other published sobemoviruses. These results suggest that the sobemoviruses could now be divided into two distinct subgroups based on those that express the RNA-dependent RNA polymerase from a single, in-frame polyprotein, and those that express it via a -1 translational frameshifting mechanism.

Rice ragged stunt oryzavirus genome segments S7 and S10 encode non-structural proteins of M(r) 68 025 (Pns7) and M(r) 32364 (Pns10) : brief report

The nucleotide sequences of genome segments S7 and S10 of a Thai-isolate of rice ragged stunt virus (RRSV) were determined. The 1938 bp S7 sequence contains a single large open reading frame (ORF) spanning nucleotides 20 to 1 843 that is predicted to encode a protein of M(r) 68 025. The 1 162 bp S10 sequence has a major ORF spanning nucleotides 142 to 1 032 that is predicted to encode a protein of M(r) 32364. This S10 ORF is preceded by a small ORF (nt 20-55) which is probably a minicistron. Coupled in vitro transcription-translation from the two major ORFs gave protein products of the expected sizes. However, no protein was visualised from S10 when the small ORF sequence was included. Proteins were expressed in Escherichia coli from the full length ORF of S7 (P7) and from a segment of the S10 ORF (P10) fused to the ORF of glutathione S-transferase (GST). Neither fusion protein was recognised by polyclonal antibodies raised against RRSV particles. Furthermore, polyclonal antibodies raised against GST-P7 fusion protein did not recognise any virion structural polypeptides. These data strongly suggest that the proteins P7 and P10 do not form part of RRSV particle. This is further supported by observed sequence homology (though very weak) of predicted.

Genome segment 5 of rice ragged stunt virus encodes a virion protein

The complete nucleotide sequence of the genome segment 5 (S5) of a Thai isolate of rice ragged stunt virus (RRSV) was determined. The 2682 nucleotide sequence contains a single long open reading frame capable of encoding a polypeptide with a molecular mass of ~91 kDa. Polypeptides encoded by various truncated cDNAs of S5 were expressed using the pGEX fusion protein vector and the highest level of fusion protein was obtained from a construct encoding a hydrophilic region of S5 protein. Antibodies raised against this fusion protein recognized a minor polypeptide, with a molecular mass of ~ 91 kDa, that was present in purified preparations of RRSV particles, infected insect vectors and infected rice plants. This indicates that RRSV S5 encodes a minor structural protein. Comparing the RRSV S5 sequence with sequences of other reo-viruses did not reveal any significant sequence similarities.

Fine tuning of elasticity via crosslinking collagen-based materials to mediate mechanotransduction and stability using corona treatment

The common goal of tissue engineering is to develop substitutes that can closely mimic the structure of extracellular matrix (ECM). However, similarly important is the intensive material properties which have often been overlooked, in particular, for soft tissues that are not to bear load assumingly. The mechanostructural properties determine not only the structural stability of biomaterials but also their physiological functionality by directing cellular activity and regulating cell fate decision. The aim here is to emphasize that cells could sense intensive material properties like elasticity and reside, proliferate, migrate and differentiate accordinglyno matter if the construct is from a natural source like cartilage, skin etc. or of synthetic one. Meanwhile, the very objective of this work is to provide a tunable scheme for manipulating the elasticity of collagen-based constructs to be used to demonstrate how to engineer cell behavior and regulate mechanotransduction. Articular cartilage was chosen as it represents one of the most complex hierarchical arrangements of collagen meshwork in both connective tissues and ECM-like biomaterials. Corona discharge treatment was used to produce constructs with varying density of crosslinked collagen and stiffness accordingly. The results demonstrated that elastic modulus increased up to 33% for samples treated up to one minute as crosslink density was found to increase with exposure time. According to the thermal analysis, longer exposure to corona increased crosslink density as the denaturation enthalpy increased. However the spectroscopy results suggested that despite the stabilization of the collagen structure the integrity of the triple helical structure remained intact. The in vitro superficial culture of heterologous chondrocytes also determined that the corona treatment can modulate migration with increased focal adhesion of cells due to enhanced stiffness, without cytotoxicity effects, and providing the basis for reinforcing three-dimensional collagen-based biomaterials in order to direct cell function and mediate mechanotransduction.

Influence of stop position on spectacle magnification

Purpose To develop and use equations of spectacle magnification when the limiting stop is either the entrance pupil of the eye or an artificial pupil in front of a lens. Methods Spectacle magnification was determined for ophthalmic lenses in air and for water environments. The reference was the retinal image for an uncorrected eye in air with a natural pupil. Results When an artificial pupil is placed in front of lenses, spectacle magnification is hardly affected by lens power, unlike the usual situation where the natural pupil is used. The water environment provides interesting influences in which spectacle magnification is highly sensitive to the distance between the cornea and eye entrance pupil. In water, retinal images are approximately 18% bigger than in air. Wearing air-filled goggles in water increases retinal image size by about 13% compared with that when they are not worn. Conclusions The equations extend earlier understanding of spectacle magnification and should be useful for those wishing to determine magnification of ophthalmic lens systems when artificial pupils and environments such as water are used.