Gene expression variation between distinct areas of breast cancer measured from paraffin-embedded tissue cores

BACKGROUND: Diagnosis and prognosis in breast cancer are mainly based on histology and immunohistochemistry of formalin-fixed, paraffin-embedded (FFPE) material. Recently, gene expression analysis was shown to elucidate the biological variance between tumors and molecular markers were identified that led to new classification systems that provided better prognostic and predictive parameters. Archived FFPE samples represent an ideal source of tissue for translational research, as millions of tissue blocks exist from routine diagnostics and from clinical studies. These should be exploited to provide clinicians with more accurate prognostic and predictive information. Unfortunately, RNA derived from FFPE material is partially degraded and chemically modified and reliable gene expression measurement has only become successful after implementing novel and optimized procedures for RNA isolation, demodification and detection. METHODS: In this study we used tissue cylinders as known from the construction of tissue microarrays. RNA was isolated with a robust protocol recently developed for RNA derived from FFPE material. Gene expression was measured by quantitative reverse transcription PCR. RESULTS: Sixteen tissue blocks from 7 patients diagnosed with multiple histological subtypes of breast cancer were available for this study. After verification of appropriate localization, sufficient RNA yield and quality, 30 tissue cores were available for gene expression measurement on TaqMan(R) Low Density Arrays (16 invasive ductal carcinoma (IDC), 8 ductal carcinoma in situ (DCIS) and 6 normal tissue), and 14 tissue cores were lost. Gene expression values were used to calculate scores representing the proliferation status (PRO), the estrogen receptor status and the HER2 status. The PRO scores measured from entire sections were similar to PRO scores determined from IDC tissue cores. Scores determined from normal tissue cores consistently revealed lower PRO scores than cores derived from IDC or DCIS of the same block or from different blocks of the same patient. CONCLUSION: We have developed optimized protocols for RNA isolation from histologically distinct areas. RNA prepared from FFPE tissue cores is suitable for gene expression measurement by quantitative PCR. Distinct molecular scores could be determined from different cores of the same tumor specimen.

"Coaching soccer is a man’s job!" – The influence of gender stereotypes on structures for recruiting female coaches in soccer clubs

Statistics published by the German Football Association indicate that women are significantly under-represented amongst soccer coaches, especially in the amateur leagues. In this paper we analyse how gender stereotypes influence the structural conditions surrounding the recruitment of coaches to soccer clubs that contribute to the exclusion of women from coaching positions. A qualitative study of five selected soccer clubs which are members of a German regional soccer association reveals that informal and personal decision-making practices still play a fundamental role in the recruitment of coaches. These practices undermine formal guidelines and give disproportionate weight to stereotyped preferences in the recruitment of soccer coaches. In this context, decision-makers' (functionaries') professed support of women largely serves a symbolic purpose and creates an illusion of equality concerning decisions that have already been taken at an informal level.

Tn6198, a novel transposon containing the trimethoprim resistance gene dfrG embedded into a Tn916 element in Listeria monocytogenes

OBJECTIVES: To characterize Tn6198, a novel conjugative transposon from the clinical Listeria monocytogenes strain TTH-2007, which contains the tetracycline and trimethoprim resistance genes tet(M) and dfrG, respectively, and to assess its transferability in vitro and in situ. METHODS: The complete sequence of Tn6198 was determined using a primer walking strategy. Horizontal gene transfer studies were performed by filter matings, as well as on the surface of smear-ripened cheese and smoked salmon. The presence of Tn916-like circular intermediates was determined by PCR. Antibiotic resistance was determined by the broth microdilution method and microarray hybridization. RESULTS: Sequencing of Tn6198 revealed that a 3.3 kb fragment containing dfrG was integrated between open reading frames 23 and 24 of Tn916. Furthermore, an additional copy of Tn916 was present in L. monocytogenes TTH-2007. Both elements were transferred simultaneously and separately in vitro to recipients L. monocytogenes 10403S and Enterococcus faecalis JH2-2 by conjugation, resulting in either tetracycline- and trimethoprim-resistant or solely tetracycline-resistant transconjugants. On the surface of cheese and salmon, only L. monocytogenes 10403S transconjugants were detected. CONCLUSIONS: This study reports the first Tn916-like element associated with a trimethoprim resistance gene, as well as the first fully characterized transposon conferring multidrug resistance in L. monocytogenes. This is of concern, as trimethoprim is administered to listeriosis patients with β-lactam allergy and as Tn6198 has a large potential for dissemination, indicated by both intra-species and inter-genus transfer.

Does work stress make you shorter? An ambulatory field study of daily work stressors, job control, and spinal shrinkage

Body height decreases throughout the day due to fluid loss from the intervertebral disk. This study investigated whether spinal shrinkage was greater during workdays compared with nonwork days, whether daily work stressors were positively related to spinal shrinkage, and whether job control was negatively related to spinal shrinkage. In a consecutive 2-week ambulatory field study, including 39 office employees and 512 days of observation, spinal shrinkage was measured by a stadiometer, and calculated as body height in the morning minus body height in the evening. Physical activity was monitored throughout the 14 days by accelerometry. Daily work stressors, daily job control, biomechanical workload, and recreational activities after work were measured with daily surveys. Multilevel regression analyses showed that spinal disks shrank more during workdays than during nonwork days. After adjustment for sex, age, body weight, smoking status, biomechanical work strain, and time spent on physical and low-effort activities during the day, lower levels of daily job control significantly predicted increased spinal shrinkage. Findings add to knowledge on how work redesign that increases job control may possibly contribute to preserving intervertebral disk function and preventing occupational back pain.

Gender differences in the association of a high quality job and self-esteem over time: A multiwave study

High self-esteem often predicts job-related outcomes, such as high job satisfaction or high status. Theoretically, high quality jobs (HQJs) should be important for self-esteem, as they enable people to use a variety of skills and attribute accomplishments to themselves, but research findings are mixed. We expected reciprocal relationships between self-esteem and HQJ. However, as work often is more important for the status of men, we expected HQJ to have a stronger influence on self-esteem for men as compared to women. Conversely, task-related achievements violate gender stereotypes for women, who may need high self-esteem to obtain HQJs. In a 4-year cross-lagged panel analysis with 325 young workers, self-esteem predicted HQJ; the lagged effect from HQJ on self-esteem was marginally significant. In line with the hypotheses, the multigroup model showed a significant path only from self-esteem to HQJ for women, and from HQJ to self-esteem for men. The reverse effect was not found for women, and only marginally significant for men. Overall, although there were some indications for reciprocal effects, our findings suggest that women need high self-esteem to obtain HQJs to a greater degree than men, and that men base their self-esteem on HQJs to a greater extent than women.

Changes in situational and dispositional factors as predictors of job satisfaction

Arguably, job satisfaction is one of the most important variables with regard to work. When explaining job satisfaction, research usually focuses on predictor variables in terms of levels but neglects growth rates. Therefore it remains unclear how potential predictors evolve over time and how their development affects job satisfaction. Using multivariate latent growth modeling in a study with 1145 young workers over five years, we analyzed how well job satisfaction is predicted a) by levels of situational (i.e., job control) and dispositional (i.e., Core Self-Evaluations (CSE)) factors and b) by growth per year of these predictors. Results showed both intercepts and slopes to be related to each other, suggesting a joint growth of job control and CSE during early careers. Job satisfaction after five years was best predicted by the slopes of job control (β = .31, p < .001) and CSE (β = .34, p < .01). These findings provide further longitudinal evidence for the role of situational as well as dispositional factors for predicting job satisfaction. In addition, growth rates per year were better predictors than initial levels. Furthermore, a lack of change in job control or CSE went along with a drop in job satisfaction, implying that young workers need to perceive things to be improving in order to increase, or at least maintain, their level of job satisfaction. In terms of theory, the relative importance of levels versus changes deserves more attention. In terms of practical implications, our results suggest a double emphasis on job design (i.e., granting sufficient, and increasing, control) and on personal development (e.g., through training) so that people experience a match between both. Finally, negative associations between initial levels and growth rates suggest that people are quite successful in achieving a reasonable fit between their job characteristics and their needs and goals.