Genetic, functional, and histopathological evaluation of two C-terminal BRCA1 missense variants

Background: The vast majority of BRCA1 missense sequence variants remain uncharacterised for their possible effect on protein expression and function, and therefore are unclassified in terms of their pathogenicity. BRCA1 plays diverse cellular roles and it is unlikely that any single functional assay will accurately reflect the total cellular implications of missense mutations in this gene. Objective: To elucidate the effect of two BRCA1 variants, 5236G>C (G1706A) and 5242C>A (A1708E) on BRCA1 function, and to survey the relative usefulness of several assays to direct the characterisation of other unclassified variants in BRCA genes. Methods and Results: Data from a range of bioinformatic, genetic, and histopathological analyses, and in vitro functional assays indicated that the 1708E variant was associated with the disruption of different cellular functions of BRCA1. In transient transfection experiments in T47D and 293T cells, the 1708E product was mislocalised to the cytoplasm and induced centrosome amplification in 293T cells. The 1708E variant also failed to transactivate transcription of reporter constructs in mammalian transcriptional transactivation assays. In contrast, the 1706A variant displayed a phenotype comparable to wildtype BRCA1 in these assays. Consistent with functional data, tumours from 1708E carriers showed typical BRCA1 pathology, while tumour material from 1706A carriers displayed few histopathological features associated with BRCA1 related tumours. Conclusions: A comprehensive range of genetic, bioinformatic, and functional analyses have been combined for the characterisation of BRCA1 unclassified sequence variants. Consistent with the functional analyses, the combined odds of causality calculated for the 1706A variant after multifactorial likelihood analysis (1:142) indicates a definitive classification of this variant as "benign". In contrast, functional assays of the 1708E variant indicate that it is pathogenic, possibly through subcellular mislocalisation. However, the combined odds of 262:1 in favour of causality of this variant does not meet the minimal ratio of 1000:1 for classification as pathogenic, and A1708E remains formally designated as unclassified. Our findings highlight the importance of comprehensive genetic information, together with detailed functional analysis for the definitive categorisation of unclassified sequence variants. This combination of analyses may have direct application to the characterisation of other unclassified variants in BRCA1 and BRCA2.

Policing of road safety in Oman: Perceptions and beliefs of traffic police officers

With increasing motorisation, road safety has become a major concern within Oman. Internationally, traffic policing plays a major role in improving road safety. Within Oman, the Royal Oman Police's (ROP) Directorate General of Traffic is responsible for policing traffic laws. Many common enforcement approaches originate from culturally different jurisdictions. The ROP is a relatively young policing force and may have different operational practices. Prior to applying practices from other jurisdictions it is important to understand the beliefs and expectations within the Directorate General of Traffic. Further, there is a need for individuals to understand their role and what is expected of them. Therefore, it is important to explore the agreement between levels of the ROP to determine how strategies and expectations transfer within the organisation. Interviews were conducted with 19 police officers from various levels of the ROP. A number of themes and findings emerged. Individuals at the upper level of the traffic police had a clear knowledge of the role of the ROP, believed that traffic police know what is expected of them, are well trained in their role and can have a very positive influence on road safety. These beliefs were less certain lower within the organisations with traffic officers having little knowledge of the role of the ROP or what was expected of them, felt undertrained, and believed their peers have little positive impact on road safety. There is a need to address barriers within the ROP in order to positively impact road safety.

Cloning and gastrointestinal expression of rat hephaestin: Relationship to other iron transport proteins

The membrane-bound ceruloplasmin homolog hephaestin plays a critical role in intestinal iron absorption. The aims of this study were to clone the rat hephaestin gene and to examine its expression in the gastrointestinal tract in relation to other genes encoding iron transport proteins. The rat hephaestin gene was isolated from intestinal mRNA and was found to encode a protein 96% identical to mouse hephaestin. Analysis by ribonuclease protection assay and Western blotting showed that hephaestin was expressed at high levels throughout the small intestine and colon. Immunofluorescence localized the hephaestin protein to the mature villus enterocytes with little or no expression in the crypts. Variations in iron status had a small but nonsignificant effect on hephaestin expression in the duodenum. The high sequence conservation between rat and mouse hephaestin is consistent with this protein playing a central role in intestinal iron absorption, although its precise function remains to be determined.

An Evaluation of Inventory Management and Performance in Indian Machine Tool SMEs: An Exploratory Study

Inventory Management (IM) plays a decisive role in the enhancement of efficiency and competitiveness of manufacturing enterprises. Therefore, major manufacturing enterprises are following IM practices as a strategy to improve efficiency and achieve competitiveness. However, the spread of IM culture among Small and Medium Enterprises (SMEs) is limited due to lack of initiation, expertise and financial limitations in developed countries, leave alone developing countries. With this backdrop, this paper makes an attempt to ascertain the role and importance of IM practices and performance of SMEs in the machine tools industry of Bangalore, India. The relationship between inventory management practices and inventory cost are probed based on primary data gathered from 91 SMEs. The paper brings out that formal IM practices have a positive impact on the inventory performance of SMEs.

The potential role of a late gene expression factor, lef2, from Bombyx mori nuclear polyhedrosis virus in very late gene transcription and DNA replication

Several late gene expression factors (Lefs) have been implicated in fostering high levels of transcription from the very late gene promoters of polyhedrin and p10 from baculoviruses. We cloned and characterized from Bombyx mori nuclear polyhedrosis virus a late gene expression factor (Bmlef2) that encodes a 209-amino-acid protein harboring a Cys-rich C-terminal domain. The temporal transcription profiles of lef2 revealed a 1.2-kb transcript in both delayed early and late periods after virus infection. Transcription start site mapping identified the presence of an aphidicolin-sensitive late transcript arising from a TAAG motif located at -352 nucleotides and an aphidicolin-insensitive early transcript originating from a TTGT motif located 35 nucleotides downstream to a TATA box at -312 nucleotides, with respect to the +1 ATG of lef2. BmLef2 trans-activated very late gene expression from both polyhedrin and p10 promoters in transient expression assays. Internal deletion of the Cys-rich domain from the C-terminal region abolished the transcriptional activation. Inactivation of Lef2 synthesis by antisense lef2 transcripts drastically reduced the very late gene transcription but showed little effect on the expression from immediate early promoter. Decrease in viral DNA synthesis and a reduction in virus titer were observed only when antisense lef2 was expressed under the immediate early (ie-1) promoter. Furthermore, the antisense experiments suggested that lef2 plays a direct role in very late gene transcription.

Determination of diffusion parameters and activation energy of diffusion in V3Si phase with A15 crystal structure

Diffusion such is the integrated diffusion coefficient of the phase, the tracer diffusion coefficient of species at different temperatures and the activation energy for diffusion, are determined in V3Si phase with A15 crystal structure. The tracer diffusion coefficient of Si Was found to be negligible compared to the tracer diffusion coefficient of V. The calculated diffusion parameters will help to validate the theoretical analysis of defect structure of the phase, which plays an important role in the superconductivity.

Destinations: 3 Large Cast Plays for Young People

This book is a collection of three large-cast plays written in response to a very specific problem. My work as a teacher of drama often required me to locate a script that would somehow miraculously work for a cast of unknown number and gender, and most likely uneven skills and enthusiasm, who I hadn’t even met yet. It’s a familiar dilemma for teachers and students of drama in education contexts, at whatever level you’re teaching. I’d first addressed this creative problem with scripts such as Gate 38 (2010). I had tried using scripts that already existed, but found they required such extensive editing to suit the parameters of cast and performance duration that I may as well have been writing them myself. Even in the setting of a closed studio, in altering these plays I felt I was bending the vision of the playwright, and certainly their narrative structure, out of shape. Everyone who’s attempted to stage a performance with a large cast of students in an educational setting knows it takes time to truly connect with a play, its social contexts, themes and characters. It also takes a lot of time to get on top of the practicalities of learning, rehearsing, directing and running a performance with young people. Often the curtain goes up on something unfinished and unstable. I was looking for ways to reduce the complexity of staging a script, while maintaining the potential of this process as a site of rich, enjoyable learning. Two of the plays (Duty Free and Please Be Seated) are comprised of multiple monologues, combined with music-driven ensemble sequences. The monologues enable individuals to develop and polish their own performances, work in small groups, and cut down on the laborious detail of directing naturalistic scenes based in character interaction. The third (Australian Drama) involves a lot of duologues, meaning that its rehearsal process can happily employ that mainstay of the drama classroom: small group work. There’s plenty of room to move in terms of gender-blind casting as well. Please be Seated is mainly young women. The scripts also contain ensemble-based interludes which are non-verbal, music driven, with a choreographic element. They have also springboarded further explorations in form. The ethical and aesthetic complexities of verbatim works; the interaction between music and theatre; and meta-concerns related to the performing of performance: ‘how can the act of acting ‘acted’. The narratives of all three of these plays are deliberately open, enabling the flexible casting and on-the hop editing that large-group, time-poor processes sometimes necessitate. Duty Free is about the overseas ‘adventures’ of young people. Please Be Seated is based in verbatim text about young people falling in and out of love. Australian Drama is about young people in a drama classroom trying to connect with each other and put their own shine on dull fragments of the theatrical canon. The plays were published as a collection in hardcopy and digital editions by Playlab Press in 2015. Please be Seated is a co-write with a large group. These co-author’s names are listed in the publication, and below in ‘additional information’.

Studies On Friction And Formation Of Transfer Layer In Hcp Metals

Surface texture plays an important role in the frictional behavior and transfer layer formation of contacting surfaces. In the present investigation, basic experiments were conducted using an inclined pin-on-plate sliding apparatus to better understand the role of surface texture on the coefficient of friction and the formation of a transfer layer. In the experiments, soft HCP materials such as pure Mg and pure Zn were used for the pins and a hardened 080 M40 steel was used for the plate. Two surface parameters of the steel plates—roughness and texture—were varied in tests that were conducted at a sliding speed of 2 mm/s in ambient conditions under both dry and lubricated conditions. The morphologies of the worn surfaces of the pins and the formation of the transfer layer on the counter surfaces were observed using a scanning electron microscope. In the experiments, the occurrence of stick-slip motion, the formation of a transfer layer, and the value of friction were recorded. With respect to the friction, both adhesion and plowing components were analyzed. Based on the experimental results, the effect of surface texture on the friction was attributed to differences in the amount of plowing. Both the plowing component of friction and the amplitude of stick-slip motion were determined to increase surface textures that promote plane strain conditions and decrease the textures that favor plane stress conditions.

Glycodelin A, an immunomodulatory protein in the endometrium, inhibits proliferation and induces apoptosis in monocytic cells

Glycodelin A (GdA), is a lipocalin with an immunomodulatory role, secreted by the endometrium under progesterone regulation and proposed to play a role in protecting the fetus from maternal immune attack. Glycodelin A has an inhibitory effect on the proliferation of T cells and B cells and also on the activity of natural killer cells. We have earlier demonstrated that the inhibitory effect of glycodelin A on T cell proliferation is due to apoptosis induced in these cells through the caspase-dependent intrinsic mitochondrial pathway. Studies reported until now have shown that glycodelin modulates the adaptive immune responses. We, therefore, decided to look at its effect, if any, on the innate immune system. The effect of glycodelin on monocytes was studied using human monocytic cell lines, THP1 and U937, and primary human monocytes as model systems. We demonstrated that glycodelin inhibited the proliferation of THP1 and U937 and induced apoptosis in these cells as well as in primary monocytes. We found that this signaling was caspase-independent but followed the intrinsic mitochondrial pathway of apoptosis. No effect of glycodelin was seen on the phagocytic ability of monocytes post-differentiation into macrophages. These observations suggest that, at the fetomaternal interface, glycodelin plays a protective role by deleting the monocytes that could become pro-inflammatory. Importantly, leaving the macrophages untouched to carry on with efficient clearance of the apoptotic cells.

Synthesis of 2-(5-((5-(4-chlorophenyl)furan 2-yl)methylene)-4-oxo-2-thioxo-thiazolidi n-3-yl)acetic acid derivatives and evaluation of their cytotoxicity and induction of apoptosis in human leukemia cells

In order to explore the anticancer effect associated with the thiazolidinone framework, several 2-(5-((5-(4-chlorophenyl)furan-2-yl)methylene)-4-oxo-2-thioxothiazolidin-3-yl)acetic acid derivatives 5(a-1) were synthesized. Variation in the functional group at C-terminal of the thiazolidinone led to set of compounds bearing amide moiety. Their chemical structures were confirmed by H-1 NMR, IR and Mass Spectra analysis. These thiazolidinone compounds containing furan moiety exhibits moderate to strong antiproliferative activity in a cell cycle stage-dependent and dose dependent manner in two different human leukemia cell lines. The importance of the electron donating groups on thiazolidinone moiety was confirmed by MTT and Trypan blue assays and it was concluded that the 4th position of the substituted aryl ring plays a dominant role for its anticancer property. Among the synthesized compounds, 5e and 5f have shown potent anticancer activity on both the cell lines tested. To rationalize the role of electron donating group in the induction of cytotoxicity we have chosen two molecules (5e and 5k) having different electron donating group at different positions. LDH assay, Flow cytometric analysis and DNA fragmentation suggest that 5e is more cytotoxic and able to induce the apoptosis. (C) 2009 Elsevier Ltd. All rights reserved.

Oestrogenic regulation and differential expression of WNT4 in the bonnet monkey and rodent epididymis

A role for oestrogen in regulating fluid reabsorption in the monkey epididymis was recently demonstrated. Here, these Studies are extended to identify potential oestrogen-regulated proteins in the cauda region of monkey epididymis treated with vehicle and oestrogen receptor antagonist (ICI 182780). Two-dimensional electrophoretic analysis was used to identify the proteins. The results indicated down-regulation of WNT4 in the ICI-182780-treated monkey cauda. In addition. the Wnt4f mRNA concentration was also reduced in the caput regions of ICI-182780-treated rats and oestrogen receptor knockout mice. WNT4 is a key regulator of gonadal differentiation in humans and mice and plays a pivotal role in early mouse embryogenesis. The results of the present Study establish the presence of WNT4 in the monkey epididymis and its regulation by oestrogen, and Suggest a role for WNT4 in maintaining epididymal homeostasis.

Impact of rRNA methylations on ribosome recycling and fidelity of initiation in Escherichia coli

Ribosomal RNA (rRNA) contains a number of modified nucleosides in functionally important regions including the intersubunit bridge regions. As the activity of ribosome recycling factor (RRF) in separating the large and the small subunits of the ribosome involves disruption of intersubunit bridges, we investigated the impact of rRNA methylations on ribosome recycling. We show that deficiency of rRNA methylations, especially at positions 1518 and 1519 of 16S rRNA near the interface with the 50S subunit and in the vicinity of the IF3 binding site, adversely affects the efficiency of RRF-mediated ribosome recycling. In addition, we show that a compromise in the RRF activity affords increased initiation with a mutant tRNA(fMet) wherein the three consecutive G-C base pairs ((29)GGG(31):39CCC41), a highly conserved feature of the initiator tRNAs, were mutated to those found in the elongator tRNA(Met) ((29)UCA(31):(39)psi GA(41)). This observation has allowed us to uncover a new role of RRF as a factor that contributes to fidelity of initiator tRNA selection on the ribosome. We discuss these and earlier findings to propose that RRF plays a crucial role during all the steps of protein synthesis.

Groucho homologue Grg5 interacts with the transcription factor Runx2-Cbfa1 and modulates its activity during postnatal growth in mice

Runx2-Cbfal, a Runt transcription factor, plays important roles during skeletal development. It is required for differentiation and function of osteoblasts. In its absence, chondrocyte hypertrophy is severely impaired and there is no vascularization of cartilage templates during skeletal development. These tissue-specific functions of Runx2 are likely to be dependent on its interaction with other proteins. We have therefore searched for proteins that may modulate the activity of Runx2. The yeast two-hybrid system was used to identify a groucho homologue, Grg5, as a Runx2-interacting protein. Grg5 enhances Runx2 activity in a cell culture-based assay and by analyses of postnatal growth in mice we demonstrate that Grg5 and Runx2 interact genetically. We also show that Runx2 haploinsufficiency in the absence of Grg5 results in a more severe delay in ossification of cranial sutures and fontanels than occurs with Runx2 haploinsufficiency on a wild-type background. Finally, we find shortening of the proliferative and hypertrophic zones, and expansion of the resting zone in the growth plates of Runx2(+/-)Grg5(-/-) mice that are associated with reduced Ihh expression and Indian hedgehog (Ihh) signaling. We therefore conclude that Grg5 enhances Runx2 activity in vivo.

'Somalia lives with me' : Homeland and Religion in the Lives of Finnish Somalis

The purpose of this thesis is to examine Somalis who live in Finland through their memories. These memories are interpreted as subjective experiences emphasizing the significance of the past and religion in these memories. On the other hand, these memories are understood as a part of collective social memory of Somalis. This study also constructs a comparative perspective for practising Islam in Somalia and Finland. The methodological framework is based on theories of social memory, oral history and narrative analysis. The empirical data is collected by interviewing Somalis living in Finland. The interviews were conducted by using the method of half-structured thematic interview. The data consists of seven interviews. The interviewees are in the focus of this study since their experiences are considered as the main sources of information for this study. The empirical data of the study reveals that Somalis have maintained strong relations to Somalia. The relationship to Somalia is mainly constructed on positive memories. Memories from Somalia have acquired a significant role in the lives of the interviewees. Those memories will define their relation to both past and the present. In the context of religious memories, Islam is described as a way of living which provides advice and defines the terms of everyday life. As a part of those religious memories, the transmitting of Islamic and Somali values plays a significant role in the lives of Somalis in Finland. In such transmitting process of the values, the social religious memory has acquired a significant role. In the context of Islam in Finland, the religious education of children is mentioned as one of the most important features of the Islamic faith by the interviewees. In general, the practice of Islam does not create any major problems for the interviewees in Finland. The interviewees describe their practice of Islam quite similar when compared to their religious life in Somalia. The empirical data also points out the fact that the meaning of Islam has not changed after moving to Finland. Keywords: Somalis, Somalia, Islam, oral history, narrative research

Corrosive and abrasive wear in ore grinding

The relative significance of corrosive and abrasive wear in ore grinding is discussed. Laboratory marked ball wear tests were carried out with magnetic taconite and quartzite under different conditions, namely dry, wet and in the presence of an organic liquid. The effect of different modes of aeration and of pyrrhotite addition on the ball wear using mild steel, high carbon low alloy steel and austenitic stainless steel balls was evaluated. Results indicate that abrasive wear plays a significant role in ore grinding in the absence of sulfides, and rheological properties of the ore slurry influenced such wear. The effect of oxygen on corrosive wear becomes increasingly felt in the presence of a sulfide mineral such as pyrrhotite. Wear characteristics of the three types of ball materials under different grinding conditions are illustrated.