GATA3 : a promising marker for metastatic breast carcinoma in serous effusion specimens

Background The utility of GATA3 as a marker for metastatic breast carcinoma in serous effusion specimens was investigated. Methods Cell block sections from 74 serous effusion specimens (32 ascitic, 2 pericardial and 40 pleural fluids) were stained with an anti-GATA3 murine monoclonal antibody. The specimens included 62 confirmed metastatic carcinomas from breast (30), female genital tract (13), gastrointestinal tract (7), lung adenocarcinoma (9), pancreas (1), kidney (1) and bladder (1). The breast carcinoma cases included 15 ductal carcinomas, 8 lobular carcinomas and in 7 the histology sub-type was not available. Twelve cases containing florid reactive mesothelial cells were also stained. The breast carcinoma cases were also stained for mammaglobin and GCDFP-15 to compare sensitivity with GATA3. Results Positive nuclear staining for GATA3 was present in 90% (27/30) of metastatic breast carcinoma specimens. All non-breast metastatic carcinomas tested were negative with the exception of the single case of metastatic urothelial carcinoma. No staining was observed in any of the benign reactive cases or in benign mesothelial cells present in the malignant cell block preparations. Two cases showed weak positivity of benign lymphoid cells. Staining results were unambiguous as all positive cases showed intense nuclear staining in >50% of tumor cells. Mammaglobin (57%; 17/30) and GCDFP-15 (33%; 10/30) were less sensitive markers of breast carcinoma. If used in a panel, mammaglobin and GCFP-15 staining would have identified only one additional case to those stained with GATA3. Conclusions GATA3 may be a useful addition to immunostaining panels for serous effusion specimens when metastatic breast carcinoma is a consideration.

The MyD88+ phenotype is an adverse prognostic factor in epithelial ovarian cancer

The prognosis of epithelial ovarian cancer is poor in part due to the high frequency of chemoresistance. Recent evidence points to the Toll-like receptor-4 (TLR4), and particularly its adaptor protein MyD88, as one potential mediator of this resistance. This study aims to provide further evidence that MyD88 positive cancer cells are clinically significant, stem-like and reproducibly detectable for the purposes of prognostic stratification. Expression of TLR4 and MyD88 was assessed immunohistochemically in 198 paraffin-embedded ovarian tissues and in an embryonal carcinoma model of cancer stemness. In parallel, expression of TLR4 and MyD88 mRNA and regulatory microRNAs (miR-21 and miR-146a) was assessed, as well as in a series of chemosensitive and resistant cancer cells lines. Functional analysis of the pathway was assessed in chemoresistant SKOV-3 ovarian cancer cells. TLR4 and MyD88 expression can be reproducibly assessed via immunohistochemistry using a semi-quantitative scoring system. TLR4 expression was present in all ovarian epithelium (normal and neoplastic), whereas MyD88 was restricted to neoplastic cells, independent of tumour grade and associated with reduced progression-free and overall survival, in an immunohistological specific subset of serous carcinomas, p<0.05. MiR-21 and miR-146a expression was significantly increased in MyD88 negative cancers (p<0.05), indicating their participation in regulation. Significant alterations in MyD88 mRNA expression were observed between chemosensitive and chemoresistant cells and tissue. Knockdown of TLR4 in SKOV-3 ovarian cells recovered chemosensitivity. Knockdown of MyD88 alone did not. MyD88 expression was down-regulated in differentiated embryonal carcinoma (NTera2) cells, supporting the MyD88+ cancer stem cell hypothesis. Our findings demonstrate that expression of MyD88 is associated with significantly reduced patient survival and altered microRNA levels and suggest an intact/functioning TLR4/MyD88 pathway is required for acquisition of the chemoresistant phenotype. Ex vivo manipulation of ovarian cancer stem cell (CSC) differentiation can decrease MyD88 expression, providing a potentially valuable CSC model for ovarian cancer.

Combined marrow stromal cell-sheet techniques and high-strength biodegradable composite scaffolds for engineered functional bone grafts

In this study, cell sheets comprising multilayered porcine bone marrow stromal cells (BMSC) were assembled with fully interconnected scaffolds made from medical-grade polycaprolactone–calcium phosphate (mPCL–CaP), for the engineering of structural and functional bone grafts. The BMSC sheets were harvested from culture flasks and wrapped around pre-seeded composite scaffolds. The layered cell sheets integrated well with the scaffold/cell construct and remained viable, with mineralized nodules visible both inside and outside the scaffold for up to 8 weeks culture. Cells within the constructs underwent classical in vitro osteogenic differentiation with the associated elevation of alkaline phosphatase activity and bone-related protein expression. In vivo, two sets of cell-sheet-scaffold/cell constructs were transplanted under the skin of nude rats. The first set of constructs (554mm3) were assembled with BMSC sheets and cultured for 8 weeks before implantation. The second set of constructs (10104mm3) was implanted immediately after assembly with BMSC sheets, with no further in vitro culture. For both groups, neo cortical and well-vascularised cancellous bone were formed within the constructs with up to 40% bone volume. Histological and immunohistochemical examination revealed that neo bone tissue formed from the pool of seeded BMSC and the bone formation followed predominantly an endochondral pathway, with woven bone matrix subsequently maturing into fully mineralized compact bone; exhibiting the histological markers of native bone. These findings demonstrate that large bone tissues similar to native bone can be regenerated utilizing BMSC sheet techniques in conjunction with composite scaffolds whose structures are optimized from a mechanical, nutrient transport and vascularization perspective.

Evaluation of a hybrid scaffold/cell construct in repair of high-load-bearing osteochondral defects in rabbits

The objective of this study was to evaluate the feasibility and potential of a hybrid scaffold system in large- and high-load-bearing osteochondral defects repair. The implants were made of medical-grade PCL (mPCL) for the bone compartment whereas fibrin glue was used for the cartilage part. Both matrices were seeded with allogenic bone marrow-derived mesenchymal cells (BMSC) and implanted in the defect (4 mm diameter×5 mm depth) on medial femoral condyle of adult New Zealand White rabbits. Empty scaffolds were used at the control side. Cell survival was tracked via fluorescent labeling. The regeneration process was evaluated by several techniques at 3 and 6 months post-implantation. Mature trabecular bone regularly formed in the mPCL scaffold at both 3 and 6 months post-operation. Micro-Computed Tomography showed progression of mineralization from the host–tissue interface towards the inner region of the grafts. At 3 months time point, the specimens showed good cartilage repair. In contrast, the majority of 6 months specimens revealed poor remodeling and fissured integration with host cartilage while other samples could maintain good cartilage appearance. In vivo viability of the transplanted cells was demonstrated for the duration of 5 weeks. The results demonstrated that mPCL scaffold is a potential matrix for osteochondral bone regeneration and that fibrin glue does not inherit the physical properties to allow for cartilage regeneration in a large and high-load-bearing defect site. Keywords: Osteochondral tissue engineering; Scaffold; Bone marrow-derived precursor cells; Fibrin glue

Critical literacy finds a "Place": Writing and social action in a low-income Australian grade 2/3 classroom

In a study of socioeconomically disadvantaged children's acquisition of school literacies, a university research team investigated how a group of teachers negotiated critical literacies and explored notions of social power with elementary children in a suburban school located in an area of high poverty. Here we focus on a grade 2/3 classroom where the teacher and children became involved in a local urban renewal project and on how in the process the children wrote about place and power. Using the students' concerns about their neighborhood, the teacher engaged her class in a critical literacy project that not only involved a complex set of literate practices but also taught the children about power and the possibilities for local civic action. In particular, we discuss examples of children's drawing and writing about their neighborhoods and their lives. We explore how children's writing and drawing might be key elements in developing "critical literacies" in elementary school settings. We consider how such classroom writing can be a mediator of emotions, intellectual and academic learning, social practice, and political activism.

Investigation of the expression of the EphB4 receptor tyrosine kinase in prostate carcinoma

Background: The EphB4 receptor tyrosine kinase has been reported as increased in tumours originating from several different tissues and its expression in a prostate cancer xenograft model has been reported. Methods: RT-PCR, western blotting and immunohistochemical techniques were used to examine EphB4 expression and protein levels in human prostate cancer cell lines LNCaP, DU145 and PC3. Immunohistochemistry was also used to examine localisation of EphB4 in tissue samples from 15 patients with prostate carcinomas. Results: All three prostate cancer cell lines expressed the EphB4 gene and protein. EphB4 immunoreactivity in vivo was significantly greater in human prostate cancers as compared with matched normal prostate epithelium and there appeared to be a trend towards increased expression with higher grade disease. Conclusions: EphB4 is expressed in prostate cancer cell lines with increased expression in human prostate cancers when compared with matched normal tissue. EphB4 may therefore be a useful anti-prostate cancer target. © 2005 Lee et al., licensee BioMed Central Ltd.

Experimental studies of wireless communication and GNSS kinematic positioning performance in high-mobility vehicle environments

In order to support intelligent transportation system (ITS) road safety applications such as collision avoidance, lane departure warnings and lane keeping, Global Navigation Satellite Systems (GNSS) based vehicle positioning system has to provide lane-level (0.5 to 1 m) or even in-lane-level (0.1 to 0.3 m) accurate and reliable positioning information to vehicle users. However, current vehicle navigation systems equipped with a single frequency GPS receiver can only provide road-level accuracy at 5-10 meters. The positioning accuracy can be improved to sub-meter or higher with the augmented GNSS techniques such as Real Time Kinematic (RTK) and Precise Point Positioning (PPP) which have been traditionally used in land surveying and or in slowly moving environment. In these techniques, GNSS corrections data generated from a local or regional or global network of GNSS ground stations are broadcast to the users via various communication data links, mostly 3G cellular networks and communication satellites. This research aimed to investigate the precise positioning system performances when operating in the high mobility environments. This involves evaluation of the performances of both RTK and PPP techniques using: i) the state-of-art dual frequency GPS receiver; and ii) low-cost single frequency GNSS receiver. Additionally, this research evaluates the effectiveness of several operational strategies in reducing the load on data communication networks due to correction data transmission, which may be problematic for the future wide-area ITS services deployment. These strategies include the use of different data transmission protocols, different correction data format standards, and correction data transmission at the less-frequent interval. A series of field experiments were designed and conducted for each research task. Firstly, the performances of RTK and PPP techniques were evaluated in both static and kinematic (highway with speed exceed 80km) experiments. RTK solutions achieved the RMS precision of 0.09 to 0.2 meter accuracy in static and 0.2 to 0.3 meter in kinematic tests, while PPP reported 0.5 to 1.5 meters in static and 1 to 1.8 meter in kinematic tests by using the RTKlib software. These RMS precision values could be further improved if the better RTK and PPP algorithms are adopted. The tests results also showed that RTK may be more suitable in the lane-level accuracy vehicle positioning. The professional grade (dual frequency) and mass-market grade (single frequency) GNSS receivers were tested for their performance using RTK in static and kinematic modes. The analysis has shown that mass-market grade receivers provide the good solution continuity, although the overall positioning accuracy is worse than the professional grade receivers. In an attempt to reduce the load on data communication network, we firstly evaluate the use of different correction data format standards, namely RTCM version 2.x and RTCM version 3.0 format. A 24 hours transmission test was conducted to compare the network throughput. The results have shown that 66% of network throughput reduction can be achieved by using the newer RTCM version 3.0, comparing to the older RTCM version 2.x format. Secondly, experiments were conducted to examine the use of two data transmission protocols, TCP and UDP, for correction data transmission through the Telstra 3G cellular network. The performance of each transmission method was analysed in terms of packet transmission latency, packet dropout, packet throughput, packet retransmission rate etc. The overall network throughput and latency of UDP data transmission are 76.5% and 83.6% of TCP data transmission, while the overall accuracy of positioning solutions remains in the same level. Additionally, due to the nature of UDP transmission, it is also found that 0.17% of UDP packets were lost during the kinematic tests, but this loss doesn't lead to significant reduction of the quality of positioning results. The experimental results from the static and the kinematic field tests have also shown that the mobile network communication may be blocked for a couple of seconds, but the positioning solutions can be kept at the required accuracy level by setting of the Age of Differential. Finally, we investigate the effects of using less-frequent correction data (transmitted at 1, 5, 10, 15, 20, 30 and 60 seconds interval) on the precise positioning system. As the time interval increasing, the percentage of ambiguity fixed solutions gradually decreases, while the positioning error increases from 0.1 to 0.5 meter. The results showed the position accuracy could still be kept at the in-lane-level (0.1 to 0.3 m) when using up to 20 seconds interval correction data transmission.

The relationship between fundamental movement skills and self-reported physical activity during Finnish junior high school

Background: Previous studies have shown that fundamental movement skills (FMS) and physical activity are related. Specifically, earlier studies have demonstrated that the ability to perform a variety of FMS increases the likelihood of children participating in a range of physical activities throughout their lives. To date, however, there have not been studies focused on the development of, or the relationship between, these variables through junior high school (that is, between the ages of 13 and 15). Such studies might provide important insights into the relationships between FMS and physical activity during adolescence, and suggest ways to design more effective physical education programmes for adolescents. Purpose: The main purposes of the study are: (1) to investigate the development of the students' self-reported physical activity and FMS from Grade 7 to Grade 9, (2) to analyse the associations among the students' FMS and self-reported physical activity through junior high school, (3) to analyse whether there are gender differences in research tasks one and/or two. Participants and setting: The participants in the study were 152 Finnish students, aged 13 and enrolled in Grade 7 at the commencement of the study. The sample included 66 girls and 86 boys who were drawn from three junior high schools in Middle Finland. Research design and data collection: Both the FMS tests and questionnaires pertaining to self-reported physical activity were completed annually during a 3 year period: in August (when the participants were in Grade 7), January (Grade 8), and in May (Grade 9). Data analysis: Repeated measures multivariate analysis of variances (MANOVAs) were used to analyse the interaction between gender and time (three measurement points) in FMS test sumscores and self-reported physical activity scores. The relationships between self-reported physical activity scores and fundamental movement skill sumscores through junior high school were analysed using Structural Equation Modelling (SEM) with LISREL 8.80 software. Findings: The MANOVA for self-reported physical activity demonstrated that both genders' physical activity decreased through junior high school. The MANOVA for the FMS revealed that the boys' FMS sumscore increased whereas the girls' skills decreased through junior high school. The SEM and squared multiple correlations revealed FMS in Grades 7 and 8 as well as physical activity in Grade 9 explained FMS in Grade 9. The portion of prediction was 69% for the girls and 55% for the boys. Additionally, physical activity measured in Grade 7 and FMS measured in Grade 9 explained physical activity in Grade 9. The portion of prediction was 12% for the girls and 29% for the boys. In the boys' group, three additional paths were found; FMS in Grade 7 explained physical activity in Grade 9, physical activity in Grade 7 explained FMS in Grade 8, and physical activity in Grade 7 explained physical activity in Grade 8. Conclusions: The study suggests that supporting and encouraging FMS and physical activity are co-related and when considering combined scores there is a greater likelihood of healthy lifelong outcomes. Therefore, the conclusion can be drawn that FMS curriculum in school-based PE is a plausible way to ensure good lifelong outcomes. Earlier studies support that school physical education plays an important role in developing students FMS and is in a position to thwart the typical decline of physical activity in adolescence. These concepts are particularly important for adolescent girls as this group reflects the greatest decline in physical activity during the adolescent period.

Expression of MUC1 and MUC2 mucins in epithelial ovarian tumours

This is the first study to describe the association between expression of MUC1 and MUC2 mucins and prognosis in ovarian cancer. Paraffin sections of epithelial ovarian tumours (n=182: 29 benign, 21 low malignant potential, and 132 invasive tumours) were analysed immunohistochemically for expression of MUC1 and MUC2 mucin core proteins. Most benign, low malignant potential, and invasive tumours showed high MUC1 expression in the cytoplasm. Low cytoplasmic expression of MUC1 was a predictor for good prognosis, particularly within stage III tumours. A minority of benign epithelial tumours, but most low malignant potential and invasive non-mucinous tumours, showed high MUC1 expression on the cell membrane. High apical MUC1 reactivity was associated with non-mucinous tumours. Low expression of MUC1 in the apical membrane was associated with early stage and good outcome for invasive tumours. Most benign and low malignant potential tumours, but only a minority of invasive tumours, showed MUC2 expression. MUC2 was found in non-mucinous as well as in mucinous tumours. The presence of MUC2 was inversely associated with high tumour grade but was not associated with altered survival. These results support experimental evidence that MUC1 influences the metastatic ability of ovarian cancer.

Paclitaxel resistance and multicellular spheroid formation are induced by kallikrein-related peptidase 4 in serous ovarian cancer cells in an ascites mimicking microenvironment

High tumor kallikrein-related-peptidase 4 (KLK4) levels are associated with a poor outcome for women with serous epithelial ovarian cancer (EOC), for which peritoneal dissemination and chemoresistance are key events. To determine the role of KLK4 in these events, we examined KLK4-transfected SKOV-3 and endogenous KLK4 expressing OVCA432 cells in 3-dimensional (3D) suspension culture to mimic the ascites microenvironment. KLK4-SKOV-3 cells formed multicellular aggregates (MCAs) as seen in ascites, as did SKOV-3 cells treated with active KLK4. MCA formation was reduced by treatment with a KLK4 blocking antibody or the selective active site KLK4 sunflower trypsin inhibitor (SFTI-FCQR). KLK4-MCAs formed larger cancer cell foci in mesothelial cell monolayers than those formed by vector and native SKOV-3 cells, suggesting KLK4-MCAs are highly invasive in the peritoneal microenvironment. A high level of KLK4 is expressed by ascitic EOC cells compared to matched primary tumor cells, further supporting its role in the ascitic microenvironment. Interestingly, KLK4 transfected SKOV-3 cells expressed high levels of the KLK4 substrate, urokinase plasminogen activator (uPA), particularly in 3D-suspension, and high levels of both KLK4 and uPA were observed in patient cells taken from ascites. Importantly, the KLK4-MCAs were paclitaxel resistant which was reversed by SFTI-FCQR and to a lesser degree by the general serine protease inhibitor, Aprotinin, suggesting that in addition to uPA, other as yet unidentified substrates of KLK4 must be involved. Nonetheless, these data suggest that KLK4 inhibition, in conjunction with paclitaxel, may improve the outcome for women with serous epithelial ovarian cancer and high KLK4 levels in their tumors.

The association between motivation in school physical education and self-reported physical activity during Finnish junior high school : a self-determination theory approach

The main purpose of this longitudinal study was to investigate the role of motivational climates, perceived competence and motivational regulations as antecedents of self-reported physical activity during junior high school years. The participants included 237 Finnish students (101 girls, 136 boys) that were 13 years old at the first stage of the study. Students completed the motivational climate and perceived competence questionnaires at Grade 7, motivation towards physical education questionnaire at Grade 8, and self-reported physical activity questionnaire at Grade 9. A path analysis revealed a path from task-involving motivational climate via perceived competence and intrinsic motivation to self-reported physical activity. Perceived competence and intrinsic motivation were statistically significant mediators between task-involving motivational climate and self-reported physical activity. This finding supports the four-stage causal sequence model of motivation.

Melanocortin-1 receptor genotype is a risk factor for basal and squamous cell carcinoma

MC1R gene variants have previously been associated with red hair and fair skin color, moreover skin ultraviolet sensitivity and a strong association with melanoma has been demonstrated for three variant alleles that are active in influencing pigmentation: Arg151Cys, Arg160Trp, and Asp294His. This study has confirmed these pigmentary associations with MC1R genotype in a collection of 220 individuals drawn from the Nambour community in Queensland, Australia, 111 of whom were at high risk and 109 at low risk of basal cell carcinoma and squamous cell carcinoma. Comparative allele frequencies for nine MC1R variants that have been reported in the Caucasian population were determined for these two groups, and an association between prevalence of basal cell carcinoma, squamous cell carcinoma, solar keratosis and the same three active MC1R variant alleles was demonstrated [odds ratio = 3.15 95% CI (1.7, 5.82)]. Three other commonly occurring variant alleles: Val60Leu, Val92Met, and Arg163Gln were identified as having a minimal impact on pigmentation phenotype as well as basal cell carcinoma and squamous cell carcinoma risk. A significant heterozygote effect was demonstrated where individuals carrying a single MC1R variant allele were more likely to have fair and sun sensitive skin as well as carriage of a solar lesion when compared with those individuals with a consensus MC1R genotype. After adjusting for the effects of pigmentation on the association between MC1R variant alleles and basal cell carcinoma and squamous cell carcinoma risk, the association persisted, confirming that presence of at least one variant allele remains informative in terms of predicting risk for developing a solar-induced skin lesion beyond that information wained through observation of pigmentation phenotype.

The effects of risk factors and protective factors on influencing engagement in risky behaviours and injury experiences for high-risk adolescents

High-risk adolescents are most vulnerable to the negative outcomes of risk taking behaviour, such as injury. It has been theorised by Jessor (1987) that adolescent risk behaviours (e.g. violence, alcohol use) can be predicted by assessing the risk factors (e.g. peer models for violence) and protective factors (e.g. school connectedness) in a young person’s life. The aim of this research is to examine the influence of risk factors and protective factors on the proneness of high-risk adolescents to engage in risky behaviour. 2,521 Grade 9 students (13-14 years of age) from 35 schools in Queensland, Australia participated in this study. The findings examine the influence of risk factors and protective factors on self-reported risky behaviour and injury experiences for adolescents who have been categorized as high-risk. Thereby, providing insight that may be used to target preventive interventions aimed at high-risk adolescents.

Tracking structural development through data modelling in highly able Grade 1 students

A 3-year longitudinal study Transforming Children’s Mathematical and Scientific Development integrates, through data modelling, a pedagogical approach focused on mathematical patterns and structural relationships with learning in science. As part of this study, a purposive sample of 21 highly able Grade 1 students was engaged in an innovative data modelling program. In the majority of students, representational development was observed. Their complex graphs depicting categorical and continuous data revealed a high level of structure and enabled identification of structural features critical to this development.

Phase III trial of gemcitabine and carboplatin versus mitomycin, ifosfamide, and cisplatin or mitomycin, vinblastine, and cisplatin in patients with advanced nonsmall cell lung carcinoma

BACKGROUND. The authors compared gemcitabine and carboplatin (GC) with mitomycin, ifosfamide, and cisplatin (MIC) or mitomycin, vinblastine, and cisplatin (MVP) in patients with advanced nonsmall cell lung carcinoma (NSCLC). The primary objective was survival. Secondary objectives were time to disease progression, response rates, evaluation of toxicity, disease-related symptoms, World Health Organization performance status (PS), and quality of life (QoL). METHODS. Three hundred seventy-two chemotherapy-naïve patients with International Staging System Stage III/IV NSCLC who were ineligible for curative radiotherapy or surgery were randomized to receive either 4 cycles of gemcitabine (1000 mg/m2 on Days 1, 8, and 15) plus carboplatin (area under the serum concentration-time curve, 5; given on Day 1) every 4 weeks (the GC arm) or MIC/MVP every 3 weeks (the MIC/MVP arm). RESULTS. There was no significant difference in median survival (248 days in the MIC/MVP arm vs. 236 days in the GC arm) or time to progression (225 days in the MIC/MVP arm vs. 218 days in the GC arm) between the 2 treatment arms. The 2-year survival rate was 11.8% in the MIC/MVP arm and 6.9% in the GC arm. The 1-year survival rate was 32.5% in the MIC/MVP arm and 33.2% in the GC arm. In the MIC/MVP arm, 33% of patients responded (4 complete responses [CRs] and 57 partial responses [PRs]) whereas in the GC arm, 30% of patients responded (3 CRs and 54 PRs). Nonhematologic toxicity was comparable for patients with Grade 3-4 symptoms, except there was more alopecia among patients in the MIC/MVP arm. GC appeared to produce more hematologic toxicity and necessitated more transfusions. There was no difference in performance status, disease-related symptoms, of QoL between patients in the two treatment arms. Fewer inpatient stays for complications were required with GC. CONCLUSIONS. The results of the current study failed to demonstrate any difference in efficacy between the newer regimen of GC and the older regimens of MIC and MVP. © 2003 American Cancer Society.