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The Spatial-Numerical Association of Response Codes (SNARC) effect - i.e., faster responses to small numbers with the left compared to the right side and to large numbers with the right compared to the left side - suggests that numbers are associated with space. However, it remains unclear whether the SNARC effect evolves from a number's magnitude or the ordinal position of a number in working memory. One problem is that, in different paradigms, the task demands influence the role of ordinality and magnitude. While single-task setups in which participants judge the parity of a displayed number indicate the importance of magnitude for the SNARC effect, evidence for ordinal influences usually comes from experiments where ordinal sequences have to be memorized or setups in which participants possess pre-existing knowledge of the ordinality of stimuli. Therefore, in this preregistered study, we employed a SNARC task without secondary ordinal sequence memorization. We dissociate ordinal and magnitude accounts by carefully manipulating experimental stimulus sets. The results indicate that even though the magnitude model better accounts for the observed data, the ordinal position seems to matter as well. Hence, numbers are associated with space in both a magnitude- and an order-respective manner, yielding a mixture of both compatibility effects. Moreover, a multiple coding framework may most accurately explain the roots of the SNARC effect. [ABSTRACT FROM AUTHOR]

This study investigated the interplay between agency and therapeutic bond in predicting patient symptoms in outpatient psychotherapy. A total of N = 731 patients provided measurements of agency (Therapeutic Agency Inventory; TAI), therapeutic bond (bond subscale of Working Alliance Inventory-Short Form Revised [WAI-SR]), and symptoms (Symptom Checklist Short Form [SCL-K11]) every fifth session of long-term treatment for up to 60 sessions. When investigated in separate models, both more agency and a stronger therapeutic bond predicted symptom improvement. However, within-person changes and between-person differences in agency predicted symptom improvement over and above the effects of therapeutic bond. Multilevel response surface analysis was used to further investigate the interplay between agency and therapeutic bond. When both agency and therapeutic bond levels were high, symptoms improved the most. When agency and therapeutic bond ratings differed, symptom ratings were significantly lower when agency exceeded therapeutic bond levels than when therapeutic bond ratings exceeded agency. Findings suggest that both agency and therapeutic bond are important treatment factors, but outcome could be improved when a strong therapeutic bond is combined with an equally strong sense of agency that empowers patients to pursue changes in their lives. When a strong therapeutic bond is present, but the patient feels less agentic, therapists may want to foster agency to improve outcomes. [ABSTRACT FROM AUTHOR]

The Spatial-Numerical Association of Response Codes (SNARC) effect - i.e., faster responses to small numbers with the left compared to the right side and to large numbers with the right compared to the left side - suggests that numbers are associated with space. However, it remains unclear whether the SNARC effect evolves from a number's magnitude or the ordinal position of a number in working memory. One problem is that, in different paradigms, the task demands influence the role of ordinality and magnitude. While single-task setups in which participants judge the parity of a displayed number indicate the importance of magnitude for the SNARC effect, evidence for ordinal influences usually comes from experiments where ordinal sequences have to be memorized or setups in which participants possess pre-existing knowledge of the ordinality of stimuli. Therefore, in this preregistered study, we employed a SNARC task without secondary ordinal sequence memorization. We dissociate ordinal and magnitude accounts by carefully manipulating experimental stimulus sets. The results indicate that even though the magnitude model better accounts for the observed data, the ordinal position seems to matter as well. Hence, numbers are associated with space in both a magnitude- and an order-respective manner, yielding a mixture of both compatibility effects. Moreover, a multiple coding framework may most accurately explain the roots of the SNARC effect. [ABSTRACT FROM AUTHOR]

Background: Patients with anorexia nervosa (AN) show overgeneralization of memory (OGM) when generating autobiographical episodes related to food and body shape. These memories are central for the construction of a coherent self‐concept, interpersonal relationships, and problem‐solving abilities. The current study aims to investigate changes in autobiographical memory following weight gain. Methods: OGM was assessed with an adapted version of the Autobiographical Memory Test including food‐, body‐, depression‐related, and neutral cues. N = 41 female patients with AN (28 restricting‐, 13 binge‐eating/purging‐subtype; mean disease duration: 4.5 years; mean BMI: 14.5 kg/m2) and N = 27 healthy controls (HC) were included at baseline. After inpatient treatment (mean duration: 11 weeks), 24 patients with AN and 24 age‐matched HC were reassessed. Group differences were assessed using independent samples t‐tests for cross‐sectional comparisons and repeated measures ANOVAs for longitudinal data. Results: At baseline, patients with AN generated significantly fewer specific memories than HC, independent of word category (F(1.66) = 27.167, p < 0.001). During inpatient stay, the average weight gain of patients with AN was 3.1 body mass index points. At follow‐up, patients with AN showed a significant improvement in the number of specific memories for both depression‐related and neutral cues, but not for food‐ and body‐related cues. Conclusions: Generalised OGM (i.e., independent of word category) in patients with AN before weight restoration may be a general incapacity to recall autobiographical memory. After weight gain, the previously well‐studied pattern of eating disorder‐related OGM emerges. The clinical relevance of the continuing disorder‐related OGM in patients with AN after weight gain is discussed. Highlights: 'Overgeneral Memory' (OGM) in severely underweight patients (mean body mass index = 14.5 kg/m2) with anorexia nervosa (AN) might be a state marker in the form of a general incapacity to recall autobiographical memories, regardless of emotional valence.Reduced symptoms of depression in patients with AN after weight gain, compared with the beginning of inpatient treatment, may be an indication of an overall improvement in mental health as negative memories become more specific.The persisting eating disorder‐related OGM after weight gain might be a maintaining factor of AN facilitating relapse and could also reflect increased dysfunctional emotion processing despite decreased eating disorder symptoms: functional avoidance might be a protective mechanism in the form of less confrontation with eating disorder‐related content through generalised memories. [ABSTRACT FROM AUTHOR]

Objective: This study examined the reciprocal effects between therapeutic agency, working alliance, and symptoms during psychotherapy. We aimed to predict symptom improvement by previous changes in either agency or alliance. In addition, we examined whether alliance development was predicted by previous changes in agency. Method: A sample of 386 patients in psychodynamic outpatient psychotherapy answered the Therapeutic Agency Inventory (TAI), the Working Alliance Inventory-SR (WAI-SR), and the Symptom Checklist-K11 (SCL-K11) after Sessions 1, 5, 10, 15, and 20. Dynamic panel models were estimated using structural equation modeling. Associations were tested while controlling for autoregressive effects and differentiating within-person changes over time from between-person differences. Results: Increases in agency predicted subsequent symptom improvement. Similarly, increases in alliance predicted subsequent symptom improvement. For agency and alliance, we found a more complex pattern with varying reciprocal effects over time. Conclusions: Findings show evidence for agency and alliance as curative change factors in psychodynamic psychotherapy. The study supports the importance of both agency and alliance and further suggests that both mechanisms may need to be balanced in successful psychotherapies. Public Health Significance: This study highlights the importance of including patients as active agents of change in psychotherapy. Patients who experience higher intentional influence over the process of psychotherapeutic change show an improved symptom response. [ABSTRACT FROM AUTHOR]

Pollen sterility (PS) in mother lines is crucial for hybrid crossings in plant breeding. 21 di‐ (2x) and tetraploid (4x) accessions of the hermaphroditic chamomile were screened for spontaneous, thermic or gametocide induced PS. Two crossing trials with the diploid accessions 'Bona' (BON) and 'Hungary 2' (HUN2) should specify maternally inherited cytoplasmic male sterility (CMS). Progeny thereof showed increased PS. BON as mother (BH) entailed means of 30 and 22% PS, versus 8% and 20% after reciprocal crossings. Backcrosses of BH with HUN2 in paternal and maternal direction revealed increasing mean PS (24% and 32%), without verification of CMS. A split plot design of 100 individuals treated with four chemical gametocides analyzed PS, seed set, germination rate and whole plant damage. Tri‐iodobenzoic acid caused the highest PS (43%). Progeny of two plants (2x, 4x) with degenerated anthers without pollen, showed 100% PS either in tetraploid F1 and F2 (4% and 11%) or in diploid F2 and F3 (26% and 23%). High temperature (30/28°C day/night) decreased the male fertility to some extent. [ABSTRACT FROM AUTHOR]

Much of the research into history teaching and ethnicity concludes that the historical narratives that children from minority ethnic groups construct differ significantly from 'mainstream' or official national narratives and are often accompanied by a sense of disengagement or even alienation from the dominant history narratives taught in schools. Our research suggests that in England (or more specifically in London) the picture is more complicated than this. First, we did not find compelling evidence that students from black and minority ethnic backgrounds feel alienated or disengaged from the British history they are taught. Second, we found a surprising similarity across the narratives that different ethnic groups chose to tell us about British history, suggesting amongst other things some curriculum inertia in schools. Where differences did exist, we suggest that these can be explained as much by gender and broad cultural influences as by ethnicity. Third, whilst what students chose to include in their narratives was broadly similar across different ethnic groups, the reasons for including them did differ. Finally, and perhaps most positively, we conclude that students of all ethnic groups are keen to engage more critically with British narratives and would relish more opportunities to do so than current school curricula appear to encourage. [ABSTRACT FROM AUTHOR]

Background: Innovative teaching models promoting beneficial therapist behaviour are central to the improvement of psychotherapeutic practice. This study investigated the effects of peer role‐play (RP) and standardised patients (SP) in the context of an intervention skill training course during psychodynamic psychotherapy training. Methods: In a mixed‐method approach, effects of intervention training with RP and SP on trainees' self‐efficacy and intervention use during treatment were investigated using trainees' self‐report questionnaires, qualitative interviews and psychotherapy session observer ratings. Twenty graduate psychotherapy students (17 female; mean age 36 years) participated in an intervention skill seminar. We assessed subjective ratings of intervention competence before and after training as well as acceptance of the training format. For a subsample of 11 participants, we examined changes in frequency and competency of intervention use with session‐based observer ratings during their current outpatient treatments. Furthermore, 10 participants reflected on their training experiences in open interviews, evaluated using inductive content analysis. Findings: The trainees' self‐efficacy for the trained interventions increased significantly after training. The training was well accepted and subjectively associated with learning success. Training transfer effects were observed regarding higher frequency and competence in the domain of clarifying and mirroring interventions during participants' outpatient psychotherapy sessions. Qualitative analysis showed training effects for self‐reflection regarding intervention use, while also promoting the development of therapeutic self‐understanding. Conclusion: Our findings provide further evidence that RP and SP training can assist the development of psychotherapy skills. Hence, the regular implementation of RP and SP training during graduate psychotherapy training seems feasible. [ABSTRACT FROM AUTHOR]

Objectives: Therapeutic agency is defined as a patient's intentional influence over the process of psychotherapeutic change. However, there is a lack of conceptually sound self-report measures with adequate psychometric properties. The aim of this study was to develop and psychometrically evaluate the patient-rated Therapeutic Agency Inventory (TAI). Method: Based on the literature, we developed items related to therapeutic agency and investigated their psychometric properties in a naturalistic study with a sample of 334 psychotherapy participants. We assessed changes in TAI scores in a subsample of 58 patients over the course of inpatient psychotherapy and related TAI scores to therapeutic improvement. Results: The TAI consists of 15 items. We performed exploratory factor analyses, and the following three factors were extracted: In-session activity, therapy-related processing, and therapist-oriented passivity. Internal consistency was.84 for the total score and ranged between.73 and.80 for each of the factors. The TAI was significantly associated with other psychotherapy process factors, self-efficacy expectations, control beliefs, lower overall psychological distress, and lower depression scores. Changes in agency during psychotherapy predicted therapy outcome, even after controlling for baseline distress. Conclusions: The TAI is a reliable, valid, and change-sensitive self-report instrument that can be used to assess agency in psychotherapy. [ABSTRACT FROM AUTHOR]

Objective: The aim of this study was to investigate associations between patients' subjective agency, their observable in‐session behavior, and the patient–therapist interaction during the early phase of psychotherapy. Methods: The sample included 52 depressed patients in psychodynamic psychotherapy. After Session 5, the patients' agency and the quality of the therapeutic alliance were assessed. Based on session recordings, two independent observers rated the patients' involvement, their interpersonal behavior, and the therapists' directiveness. Results: Higher agency was associated with stronger therapeutic alliances. Patients who indicated higher agency in their therapy participated more actively in the session and showed less hostile impact messages. Patients' agency was not related to therapists' directiveness. Conclusions: Patients' sense of agency in psychotherapy was associated with more active involvement and affiliative interaction. The findings support the idea that patients need to feel capable of acting within and having an influence on their therapy to benefit from it. [ABSTRACT FROM AUTHOR]

Objective: An increased understanding of repetitive dysfunctional patterns and their relationship to an individual's life history is regarded as a key mechanism of change in insight-oriented therapies. At the same time, empirical research on the insight-outcome relationship is rare, and its generalizability is restricted by the use of a wide range of definitions and methods among studies. The authors conducted a meta-analysis to systematically examine the association between patient insight and psychotherapy outcome across a range of treatment modalities.Method: Insight was defined as patients' understanding of associations between past and present experiences, typical relationship patterns, and the relation between interpersonal challenges, emotional experience, and psychological symptoms. From 13,849 initially identified abstracts, the authors extracted 23 independent effect sizes. A random-effects meta-analysis was performed to assess the magnitude of the insight-outcome relationship. Risk of publication bias was assessed with funnel plot inspections, Egger's regression test, and Duval and Tweedie's trim-and-fill procedure as sensitivity analyses.Results: A significant, moderate correlation (r=0.31) was observed between insight and treatment outcome. Sensitivity analyses demonstrated the robustness of the results.Conclusions: The findings support the importance of insight for psychotherapy outcome. Insight may be a relevant mechanism of change across different treatment modalities. [ABSTRACT FROM AUTHOR]

Fossil energy input, energy output, and energy use- efficiency (EUE) are important indicators of the environmental effects, resource consumption and economic performance of farming systems. In this study, the energy balance (process analysis) and EUE of different organic and conventional farming systems – mixed farming, arable farming, and agroforestry (AGFS) systems – were analyzed due to their importance in Germany. The analysis was based on long-term experiments at the Scheyern Research Farm in southern Germany. The results showed that the conversion from multi-structured organic mixed farming to a specialized organic arable farming system reduced the fossil energy input in crop production only marginally (from 5.9 to 5.5 GJ ha −1 yr −1 ), but considerably decreased dry matter yield (from 5.4 to 2.5 Mg ha −1 yr −1 ), energy output (from 99 to 46 GJ ha −1 yr −1 ) and EUE (from 16.8 to 8.3). Improved management in the conventional arable farming system (with high-yielding varieties and better N management) reduced the energy input from 14.0 to 12.2 GJ ha −1 yr −1 , increased the energy output from 155 to 179 GJ ha −1 yr −1 , and elevated the EUE from 11.1 to 14.6. The establishment of AGFS with short rotation trees (without fertilization and pesticide use) led to the reduction of energy input. Based on the results, we concluded that mixed farming is one of the best ways of producing food with high EUE under the conditions of organic farming. Therefore the conversion from organic mixed farming to an organic arable farming system is not recommended. Our AGFS results were from the first stage of a long-term experiment; it showed no negative effects on DM yield and energy output and positive effects on energy-use efficiency at this stage. However, further research is needed to know the long-term influence of AGFS. [ABSTRACT FROM AUTHOR]

Background: The focus of the outlined work is the establishment of a three-dimensional lung model for various drug-screening applications. Methods: The non-small cell lung cancer (NSCLC) cell line Colo699 was cultivated as monolayer (2D) on plates for 5 days or as microtissues (3D) using a hanging-drop system for 5 and 10 days. Cells and microtissues were treated with afatinib (10-80 µM), cisplatin (100-800 µM) or vinorelbine (25-200 µM) for 24 or 48 hours (h). Cell proliferation and viability were analysed by intra-cellular adenosine triphosphate (ATP) and lactate dehydrogenase release (LDH) assays, annexin V/propidium iodide (PI) staining, and cell cycle determination. Microtissue morphology and size, as well as cell death were evaluated via phase contrast microscopy. Results: Our results demonstrate the valid determination of viability and cell death using established assays in the 3D system for drug testing. The comparison of ATP, LDH and cytometry data showed moderate (0.40) to very strong (0.99) correlations. Thereby, we observed partially significant differences in drug efficacy between microtissues and 2D cultures dependent from the applied treatment and read-out method. Altogether, microtissues developed resistance to cisplatin and vinorelbine; but remained more vulnerable to afatinib. These findings were confirmed with microscopy. Conclusion: In summary, we established an NSCLC 3D test system with multiple assays compatible for drug-testing applications of substances with different mechanisms of action. In addition, our data support the usage of microtissues as more accurate tools for drug-efficacy testing with the possibility of long-term cultivation and treatment. [ABSTRACT FROM AUTHOR]

Angiogenesis is a key process in tumor growth and progression, which is controlled by vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs). In order to better understand the prevalence and prognostic value of VEGFR1 expression in breast cancer, a tissue microarray containing >2,100 breast cancer specimens, with clinical follow-up data, was analyzed by immunohistochemistry using an antibody directed against the membrane-bound full-length receptor protein. The results demonstrated that membranous VEGFR1 staining was detected in all (5 of 5) normal breast specimens. In carcinoma specimens, membranous staining was negative in 3.1%, weak in 6.3%, moderate in 10.9%, and strong in 79.7% of the 1,630 interpretable tissues. Strong staining was significantly associated with estrogen receptor and progesterone receptor expression, but was inversely associated with advanced tumor stage (P=0.0431), high Bloom-Richardson-Ellis Score for Breast Cancer grade and low Ki67 labeling index (both P<0.0001). Cancers with moderate to strong (high) VEGFR1 expression were associated with significantly improved overall survival, as compared with tumors exhibiting negative or weak (low) expression (P=0.0015). This association was also detected in the subset of nodal-positive cancers (P=0.0018), and in the subset of 185 patients who had received tamoxifen as the sole therapy (P=0.001). In conclusion, these data indicated that membrane-bound VEGFR1 is frequently expressed in normal and cancerous breast epithelium. In addition, reduced or lost VEGFR1 expression may serve as a marker for poor prognosis in patients with breast cancer, who might not optimally benefit from endocrine therapy. [ABSTRACT FROM AUTHOR]

Objective First studies revealed overgeneral autobiographical memories in anorexia nervosa (AN) patients. The aim of the present study was to investigate frequency, generalization and valence of autobiographical memories in AN patients in response to eating disorder-related cue words. Method Autobiographical memory was examined in 21 AN patients and 21 healthy controls (HC) using a modified version of the Autobiographical Memory Test, incorporating body-related, food-related, perfectionism-related, depression-related and neutral cues. Results Anorexia nervosa patients recalled fewer and more general autobiographical memories compared with HC. For eating disorder-related cues as against neutral ones, AN patients compared with HC showed fewer memories for food-related and body-related cues, an elevated overgeneralization for food-related cues, while the valence of the retrieved memories was more negative in response to body-related cues. Discussion This study detects disorder-related autobiographical memory alterations in AN, which are intensified in response to symptom-related cues. The findings are discussed with regard to their maladaptive function in emotion regulation. Copyright © 2015 John Wiley & Sons, Ltd and Eating Disorders Association. [ABSTRACT FROM AUTHOR]

In arthropods the cuticle forms an exoskeleton with its physical and chemical properties adapted to functions of distinct skeletal elements. The cuticle of the partes incisivae (PI) in mandibles of terrestrial isopods is a composite of chitin-protein fibrils/fibres and minerals. It consists of an unmineralized tip, a middle region with organic fibrils reinforced mainly with amorphous calcium phosphate and a base region mineralized with amorphous calcium carbonate and calcite. In this study we extend our work on the structure and material properties of the incisive cuticle employing electron backscatter diffraction (EBSD), and investigate calcite orientation patterns in the PI of two terrestrial isopod species from different habitats. We trace small-scale differences in texture sharpness and calcite microstructure, and compare calcite organization and orientation patterns in the PI with those in the tergites of the same isopod species. We observe that in the PI calcite orientation, the degree of crystal alignment, and mode of crystalline domain assemblage is highly varied within short length scales. This contrasts to calcite organization in the tergite cuticle, where calcite has only one specific texture pattern. Such a large range in the variation of calcite organization has not been observed in other carbonate biological hard tissues, such as shells and teeth, where one specific texture and microstructure prevails. Thus, the investigated isopod species are able to control crystallization of the amorphous carbonate precursor in a differential way, most probably related to the function of the individual skeletal element and the animals’ behavior. [ABSTRACT FROM AUTHOR]

In terrestrial isopods the mandibles consist of a corpus carrying strong muscle tissue, and a pars incisiva (PI) that cuts dry leaves into smaller ingestible pieces. We studied the cuticle of the PI of Porcellio scaber in order to understand region-dependent differences in its ultrastructure, composition, and the resulting mechanical properties, employing several microscopic and analytical techniques as well as nanoindentation experiments. The cuticle of the incisive tip is not mineralized and consists of an unusually thick epicuticle containing thin fibrils, two subjacent cuticular layers, and a central core containing fibrils of different orientation, either longitudinal or circumferential. A thick epicuticle of the middle region just behind the tip projects long epicuticular extensions into the subjacent endocuticle, likely to prevent delamination. A distinct exocuticular layer is lacking in the middle region. Most chitin–protein fibrils within the endocuticle are oriented in parallel pointing towards the tip. Surprisingly, the middle region is mineralized by amorphous calcium phosphate (ACP) only. Near the base, ACP is successively replaced by amorphous calcium carbonate and calcite is restricted to a distal layer in the base. At the transition between middle and base, the epicuticle forms a hybrid material containing fibrils of the exocuticle. Nanoindentation experiments reveal an increase of the stiffness and hardness from the tip towards the base and significantly higher values on transversal in comparison to longitudinal planes. The results suggest that ultrastructure and composition are adapted for conveying high forces from a rather thin cutting edge to the stable base of the PI. [ABSTRACT FROM AUTHOR]

Background: Patients with Anorexia Nervosa (AN) show a moderate deficit in overall neuropsychological functioning. Since previous studies on memory performance mainly employed cross‐sectional designs, the present study aims to investigate changes in verbal memory following weight‐gain. Methods: Verbal memory was assessed with the Wechsler Memory Scale‐Revised (WMS‐R; 'logical memory'‐story‐recall‐subtest) and the California Verbal Learning Test‐II (CVLT‐II; 'verbal learning'). Included were 31 female patients with AN (18 restricting‐, 13 purging‐subtype; average disease duration: 5.1 years; average baseline BMI: 14.4 kg/m2) and 24 medication‐free normal‐weight healthy women adjusted for age at baseline (T0). In a post‐treatment assessment of approx. 6 weeks with weight increase (T1), 18 patients with AN and 20 healthy women were assessed again. Group differences in verbal memory (i.e., WMS‐R, CVLT‐II) were assessed for the baseline comparisons with a multivariate ANOVA and longitudinal data were analysed with repeated measures (RM) ANOVAs. Results: At baseline, patients with AN as compared to healthy women displayed deficits in logical memory. In the follow‐up assessment, patients with AN improved their logical memory significantly compared to healthy controls (p < 0.006). Furthermore, groups did not differ in verbal learning neither before nor after inpatient treatment. Conclusions: Enhanced logical memory in patients with AN following weight‐gain is probably due to the impaired memory as compared to healthy controls at T0. A survivorship bias could explain the improved memory performance in longitudinal data in contrast to cross‐sectional studies. Patients with AN with poorer memory performance before inpatient treatment are at higher risk to drop out and need support. Key points: Impaired logical memory before inpatient treatment improves in patients with anorexia nervosa (AN) during weight‐gain, while verbal learning seems to be intact before as well as after inpatient treatment as compared to healthy women.Drop‐out‐analyses revealed that patients with AN with poorer logical memory performance before inpatient treatment are at higher risk to drop out, and should receive further therapeutic support.Results imply, that therapeutic interventions using cognitive strategies may be overwhelming in severely ill patients with AN during early stages of the disease, while early cognitive interventions might be effective in a less sick, more motivated subgroup of patients with AN. [ABSTRACT FROM AUTHOR]

The present study investigates the coevolution of a particular male genital process and the female spermathecal ducts in a clade of stalk-eyed flies (Diptera, Diopsidae) and debates the underlying evolutionary mechanisms. The fine morphology and interaction of the male and female genitalic structures are reconstructed from serial sections of mating pairs in one of the species. It is found that the male genital process traverses the common spermathecal duct to enter the base of one of the separate spermathecal ducts during the mating. Spermatozoa and accessory secretions are not transferred through the male genital process but can be discharged only from the male gonopore near its base. A detailed morphometric study reveals low intraspecific variation and hypoallometry of the male genital process. Across 17 species studied comparatively, the lengths of the male genital process and the female common and separate spermathecal ducts are highly variable. The length of the male genital process is correlated significantly with that of the female common spermathecal duct, but not with that of the separate spermathecal ducts. Based on the combined evidence it is concluded that the male genital process and the female common spermathecal duct have coevolved, and that sexual selection by cryptic female choice constitutes a possible and parsimonious explanation for their coevolution. Alternative or additional explanations in terms of sexually antagonistic coevolution cannot be ruled out conclusively, but are not supported by the available evidence. [ABSTRACT FROM AUTHOR]

Bortezomib is a well-established treatment option for patients with multiple myeloma (MM). It is a selective and reversible inhibitor of the proteasome that is responsible for the degradation of many regulatory proteins that are involved in apoptosis, cell-cycle regulation, or transcription. Because patients with MM are prone to develop acute renal failure, we evaluated the influence of Bortezomib on renal ischemia-reperfusion injury (IRI). Mice were subjected to renal IRI by having the renal pedicles clamped for 30 mm followed by reperfusion for 3, 24, and 48 h. Mice were either pretreated with 0.5 mg/kg body wt Bortezomib or vehicle intravenously 12 h before induction of IRI. Serum creatinine and tubular necrosis were significantly increased in Bortezomib compared with vehicle-treated mice. The inflammatory response was found to be significantly decreased in Bortezomib-treated mice as reflected by a decreased infiltration of CD4+ T cells and a significantly decreased ml cytokine expression in the kidneys. In contrast, apoptosis was significantly increased in kidneys of Bortezomib-treated mice compared with vehicle-treated controls. Increased numbers of TUNEL-positive cells/mm2 and increased mRNA expression of proapoptotic factors were detected in kidneys of Bortezomib-treated mice. Of note, p21, a cell senescence marker, was also significantly increased in kidneys of Bortezomib-treated mice. In summary, we provide evidence that Bortezomib worsens the outcome of renal IRI by leading to increased apoptosis of tubular cells despite decreased infiltrating T cells and proinflammatory mediators. [ABSTRACT FROM AUTHOR]

Pathological biopsy protocols require tissue marking dye (TMD) for orientation. In some cases (e.g., close margin), additional immunohistochemical analyses can be necessary. Therefore, the correlation between the applied TMD during macroscopy and the examined TMD during microscopy is crucial for the correct orientation, the residual tumour status and the subsequent therapeutic regime. In this context, our group observed colour changes during routine immunohistochemistry. Tissue specimens were marked with various TMD and processed by two different methods. TMD (blue, red, black, yellow and green) obtained from three different providers (A, B and C, and Whiteout/Tipp-Ex®) were used. Immunohistochemistry was performed manually via stepwise omission of reagents to identify the colour changing mechanism. Blue colour from provider A changed during immunohistochemistry into black, when 3,3′-Diaminobenzidine-tetrahydrochloride-dihydrate (DAB) and H2O2 was applied as an immunoperoxidase-based terminal colour signal. No other applied reagents, nor tissue texture or processing showed any influence on the colour. The remaining colours from provider A and the other colours did not show any changes during immunohistochemistry. Our results demonstrate an interesting and important pitfall in routine immunohistochemistry-based diagnostics that pathologists should be aware of. Furthermore, the chemical rationale behind the observed misleading colour change is discussed. [ABSTRACT FROM AUTHOR]

Huber J, Fröhlich-Reiterer EE, Sudi K, Suppan E, Weinhandl G, Jasser-Nitsche H, Aigner R, Borkenstein MH. The influence of physical activity on ghrelin and IGF-1/IGFBP-3 levels in children and adolescents with type 1 diabetes mellitus. Objectives: The aim of the study was to determine the influence of regular physical activity on ghrelin and IGF-1/IGFBP-3 levels during a diabetes camp. Methods: Twenty-eight children and adolescents (14 boys; mean age 12.1 yr) with type 1 diabetes mellitus (T1DM, mean duration of diabetes 4.8 yr) attending a 2-wk diabetes camp that features increased regular physical activities have been studied. Serum levels of ghrelin (total and acylated), growth hormone (GH), insulin-like growth factor-1 (IGF-1), insulin-like growth factor-bindng protein-3 (IGFBP-3) and insulin were measured in fasting state on day 1 and day 14. Improvement of metabolic control was documented by haemoglobin A1c (HbA1c). Glucose levels and insulin doses were determined daily. Results: Mean insulin dosage decreased from 0.87 to 0.78 U/kg/d, mean HbA1c levels decreased from 8.6 to 8.3%, but the changes were not statistical. There was a significant decline in total ghrelin. IGFBP-3 and IGF-1 decreased also significantly. Total basal ghrelin was inversely related to the change in IGFBP-3. Conclusions: We hypothesize an association between ghrelin and metabolic control in T1DM. Higher ghrelin levels might be associated with poor metabolic control. The dynamic of IGFBP-3 levels appears to be under the influence of basal ghrelin concentrations in T1DM. [ABSTRACT FROM AUTHOR]

Hintergrund: Psychodynamische Psychotherapien spielen eine wichtige Rolle im deutschen Gesundheitssystem. Der Nachweis ihrer Lehrbarkeit bekommt vor dem Hintergrund des neuen Psychotherapiestudiums eine neue Bedeutung. Dies bezieht sich insbesondere auf erfahrungsbasierte di-daktische Modelle und auf das spezifische Üben von Interventionen. Hier ist bisher unklar, welche Aspekte kompetenz-orientierter psychodynamischer Lehre mit dem subjektiven Lernerfolg zusammenhängen. Methode: In einer Stichprobe mit 47 Kandidatinnen und Kandidaten eines Ausbildungsinstitutes wurden psychodynamische Interventionen mit -Videofeedback, Peer-Rollenspielen und Simulationspatientinnen und -patienten unterrichtet. Der subjektive Kompetenzgewinn von Prä zu Post wurde in Bezug auf die Interventionen Klarifizieren, Konfrontieren und Deuten mithilfe von t-Tests abgebildet. Zudem wurden in einer multiplen linearen Regression verschiedene Prädiktoren für den subjektiven Lernerfolg getestet. Ergebnisse: Die subjektive Kompetenz nahm für die Interventionen Klarifizieren und Konfrontieren mit großen Effekten, für Deuten mit einem kleinen bis -mittleren Effekt zu. Der Lernerfolg war korreliert mit einem antizipierten Transfer in den therapeutischen Alltag, einem positiven Erleben der Arbeit mit den Simulationspatientinnen und -patienten sowie mit hilfreich erlebtem Feedback durch die Dozentinnen und Dozenten. Schlussfolgerungen: Psychodynamische Interventionen sind mit erfahrungsbasierten und kompetenzorientierten Unterrichtsformaten lehrbar. Dabei ist wichtig, dass praxisorientierte Formate eine Lernumgebung schaffen, in der die Arbeit mit Simulationspatientinnen und -patienten und das Feedback als konstruktiv und hilfreich erlebt werden. Zukünftige Untersu-chungen sollten den Transfer in den Alltag sowie longitudinale Curricula in den Fokus nehmen. [ABSTRACT FROM AUTHOR]

The transcription factor FOXO3 is associated with poor outcome in high-stage neuroblastoma (NB), as it facilitates chemoprotection and tumor angiogenesis. In other tumor entities, FOXO3 stimulates metastasis formation, one of the biggest challenges in the treatment of aggressive NB. However, the impact of FOXO3 on the metastatic potential of neuronal tumor cells remains largely unknown. In the present study, we uncover the small leucine-rich proteoglycan family member lumican (LUM) as a FOXO3-regulated gene that stimulates cellular migration in NB. By a drug-library screen we identified the small molecular weight compound repaglinide (RPG) as a putative FOXO3 inhibitor. Here, we verify that RPG binds to the FOXO3-DNA-binding-domain (DBD) and thereby silences the transcriptional activity of FOXO3. Consistent with the concept that the FOXO3/LUM axis enhances the migratory capacity of aggressive NB cells, we demonstrate that stable knockdown of LUM abrogates the FOXO3-mediated increase in cellular migration. Importantly, FOXO3 inhibition by RPG represses the binding of FOXO3 to the LUM promoter, inhibits FOXO3-mediated LUM RNA and protein expression, and efficiently abrogates FOXO3-triggered cellular “wound healing” as well as spheroid-based 3D-migration. Thus, silencing the FOXO3/LUM axis by the FDA-approved compound RPG represents a promising strategy for novel therapeutic interventions in NB and other FOXO3-dependent tumors. [ABSTRACT FROM AUTHOR]

Biomass equations for tree species and the early stages of growth used in short-rotation coppices and agroforestry systems are still lacking. Further, discussion about the structure and parameters of biomass equations are still ongoing. Yield estimations should be precise, while keeping efforts low. To determine the influence of tree species, farming system, and tree position (inner and outer row) on allometric relationships, we derived biomass equations for various tree species from organic and conventional silvoarable agroforestry systems with an alley-cropping configuration. The allometric equations were based on the power relationship between aboveground dry biomass and stem base diameter (SBD) as a single variable or in combination with tree height (H) and were calculated by log-linear mixed-effect regression. Equations span the third and fourth growth year of the first rotation and were validated on the fourth year. Neither farming system nor row position influenced allometric relationship, although biometric variables varied between trees from inner and outer rows. A general model across species explained 95% ($$R_{\text{cond}}^{2}$$Rcond2) with H as covariate. Yet, for the sake of precision, species-specific equations were necessary. The best fitting equation with only SBD as predictor had species-specific allometric factors and a general exponent across species. However, predicted yields were biased by 8–31%. Thus, functions incorporating H are recommended, as compensation for variances in height-diameter relationships due to the ontogenetic stage, site differences, or social status of the tree reduced the bias of biomass estimation (<10%).Biomass equations for tree species and the early stages of growth used in short-rotation coppices and agroforestry systems are still lacking. Further, discussion about the structure and parameters of biomass equations are still ongoing. Yield estimations should be precise, while keeping efforts low. To determine the influence of tree species, farming system, and tree position (inner and outer row) on allometric relationships, we derived biomass equations for various tree species from organic and conventional silvoarable agroforestry systems with an alley-cropping configuration. The allometric equations were based on the power relationship between aboveground dry biomass and stem base diameter (SBD) as a single variable or in combination with tree height (H) and were calculated by log-linear mixed-effect regression. Equations span the third and fourth growth year of the first rotation and were validated on the fourth year. Neither farming system nor row position influenced allometric relationship, although biometric variables varied between trees from inner and outer rows. A general model across species explained 95% ($$R_{\text{cond}}^{2}$$Rcond2) with H as covariate. Yet, for the sake of precision, species-specific equations were necessary. The best fitting equation with only SBD as predictor had species-specific allometric factors and a general exponent across species. However, predicted yields were biased by 8–31%. Thus, functions incorporating H are recommended, as compensation for variances in height-diameter relationships due to the ontogenetic stage, site differences, or social status of the tree reduced the bias of biomass estimation (<10%).Biomass equations for tree species and the early stages of growth used in short-rotation coppices and agroforestry systems are still lacking. Further, discussion about the structure and parameters of biomass equations are still ongoing. Yield estimations should be precise, while keeping efforts low. To determine the influence of tree species, farming system, and tree position (inner and outer row) on allometric relationships, we derived biomass equations for various tree species from organic and conventional silvoarable agroforestry systems with an alley-cropping configuration. The allometric equations were based on the power relationship between aboveground dry biomass and stem base diameter (SBD) as a single variable or in combination with tree height (H) and were calculated by log-linear mixed-effect regression. Equations span the third and fourth growth year of the first rotation and were validated on the fourth year. Neither farming system nor row position influenced allometric relationship, although biometric variables varied between trees from inner and outer rows. A general model across species explained 95% ($$R_{\text{cond}}^{2}$$Rcond2) with H as covariate. Yet, for the sake of precision, species-specific equations were necessary. The best fitting equation with only SBD as predictor had species-specific allometric factors and a general exponent across species. However, predicted yields were biased by 8–31%. Thus, functions incorporating H are recommended, as compensation for variances in height-diameter relationships due to the ontogenetic stage, site differences, or social status of the tree reduced the bias of biomass estimation (<10%).Biomass equations for tree species and the early stages of growth used in short-rotation coppices and agroforestry systems are still lacking. Further, discussion about the structure and parameters of biomass equations are still ongoing. Yield estimations should be precise, while keeping efforts low. To determine the influence of tree species, farming system, and tree position (inner and outer row) on allometric relationships, we derived biomass equations for various tree species from organic and conventional silvoarable agroforestry systems with an alley-cropping configuration. The allometric equations were based on the power relationship between aboveground dry biomass and stem base diameter (SBD) as a single variable or in combination with tree height (H) and were calculated by log-linear mixed-effect regression. Equations span the third and fourth growth year of the first rotation and were validated on the fourth year. Neither farming system nor row position influenced allometric relationship, although biometric variables varied between trees from inner and outer rows. A general model across species explained 95% ($$R_{\text{cond}}^{2}$$Rcond2) with H as covariate. Yet, for the sake of precision, species-specific equations were necessary. The best fitting equation with only SBD as predictor had species-specific allometric factors and a general exponent across species. However, predicted yields were biased by 8–31%. Thus, functions incorporating H are recommended, as compensation for variances in height-diameter relationships due to the ontogenetic stage, site differences, or social status of the tree reduced the bias of biomass estimation (<10%).Biomass equations for tree species and the early stages of growth used in short-rotation coppices and agroforestry systems are still lacking. Further, discussion about the structure and parameters of biomass equations are still ongoing. Yield estimations should be precise, while keeping efforts low. To determine the influence of tree species, farming system, and tree position (inner and outer row) on allometric relationships, we derived biomass equations for various tree species from organic and conventional silvoarable agroforestry systems with an alley-cropping configuration. The allometric equations were based on the power relationship between aboveground dry biomass and stem base diameter (SBD) as a single variable or in combination with tree height (H) and were calculated by log-linear mixed-effect regression. Equations span the third and fourth growth year of the first rotation and were validated on the fourth year. Neither farming system nor row position influenced allometric relationship, although biometric variables varied between trees from inner and outer rows. A general model across species explained 95% ($$R_{\text{cond}}^{2}$$Rcond2) with H as covariate. Yet, for the sake of precision, species-specific equations were necessary. The best fitting equation with only SBD as predictor had species-specific allometric factors and a general exponent across species. However, predicted yields were biased by 8–31%. Thus, functions incorporating H are recommended, as compensation for variances in height-diameter relationships due to the ontogenetic stage, site differences, or social status of the tree reduced the bias of biomass estimation (<10%).Biomass equations for tree species and the early stages of growth used in short-rotation coppices and agroforestry systems are still lacking. Further, discussion about the structure and parameters of biomass equations are still ongoing. Yield estimations should be precise, while keeping efforts low. To determine the influence of tree species, farming system, and tree position (inner and outer row) on allometric relationships, we derived biomass equations for various tree species from organic and conventional silvoarable agroforestry systems with an alley-cropping configuration. The allometric equations were based on the power relationship between aboveground dry biomass and stem base diameter (SBD) as a single variable or in combination with tree height (H) and were calculated by log-linear mixed-effect regression. Equations span the third and fourth growth year of the first rotation and were validated on the fourth year. Neither farming system nor row position influenced allometric relationship, although biometric variables varied between trees from inner and outer rows. A general model across species explained 95% ($$R_{\text{cond}}^{2}$$Rcond2) with H as covariate. Yet, for the sake of precision, species-specific equations were necessary. The best fitting equation with only SBD as predictor had species-specific allometric factors and a general exponent across species. However, predicted yields were biased by 8–31%. Thus, functions incorporating H are recommended, as compensation for variances in height-diameter relationships due to the ontogenetic stage, site differences, or social status of the tree reduced the bias of biomass estimation (<10%). [ABSTRACT FROM AUTHOR]

A letter to the editor is presented which describes four cases of cervical lymphadenitis caused by Mycobacterium bohemicum.

Background: Anorexia nervosa (AN) is characterized by an overgeneralization of food/body-related autobiographical memories (AM). This is regarded as an emotion regulation strategy with adverse long-term effects implicated in disorder maintenance and treatment resistance. Therefore, we aimed to examine neural correlates of food/body-related AM-recall in AN. Methods: Twenty-nine female patients with AN and 30 medication-free age-sex-matched normal-weight healthy controls (HC) underwent functional magnetic resonance imaging while recalling AMs in response to food/body-related and neutral cue words. To control for general knowledge retrieval, participants engaged in a semantic generation and riser detection task. Results: In comparison to HC, patients with AN generated fewer and less specific AMs in response to food/body-related words, but not for neutral cue words. Group comparisons revealed reduced activation in regions associated with self-referential processing and memory retrieval (precuneus and angular gyrus) during the retrieval of specific food/body-related AM in patients with AN. Brain connectivity in regions associated with memory functioning and executive control was reduced in patients with AN during the retrieval of specific food/body-related AM. Finally, resting-state functional connectivity analysis revealed no differences between groups, arguing against a general underlying disconnection of brain networks implicated in memory and emotional processing in AN. Conclusions: These results indicate impaired neural processing of food/body-related AM in AN, with a reduced involvement of regions involved in self-referential processing. Our findings are discussed as possible neuronal correlates of emotional avoidance in AN and provide new insights of AN-pathophysiology underscoring the importance of targeting dysfunctional emotion regulation strategies during treatment. [ABSTRACT FROM AUTHOR]

Objective: Depressive disorders are among the most common psychiatric disorders. For severely depressed patients, day clinic and inpatient settings represent important treatment options. However, little is known about patients' perceptions of the different levels of care. This study aimed to obtain an in-depth analysis of depressive patients' experiences of day clinic and inpatient treatment in a combined clinical setting. Methods: Following a randomized controlled trial comparing day clinic and inpatient psychotherapy for depression (Dinger et al. in Psychother Psychosom 83:194-195, 2014), a sample of depressive patients (n = 35) was invited to participate in a semi-structured interview during an early follow up 4 weeks after discharge. A qualitative analysis of interview transcripts was performed following the principles of constructivist thematic analysis. Results: Following analysis, 1355 single codes were identified from which five main categories and 26 themes were derived for both groups. In regard to patient group integration and skill transfer to everyday life, distinct differences could be observed between the day clinic and inpatient group. Conclusion: While adjustment to therapeutic setting and patient group integration seem to be facilitated by inpatient treatment, the day clinical setting appears to promote treatment integration into patients' everyday contexts, aiding treatment-related skill transfer to everyday life as well as alleviating discharge from clinic treatment. Further studies on depressive subject groups in day clinic and inpatient treatment should investigate aspects of group cohesion and treatment integration in relation to therapeutic outcome. [ABSTRACT FROM AUTHOR]

Introduction The interaction between the immune system and malignant diseases is a proven key target for cancer therapy. We describe an innovative 3D cell culture system comprising both immune and cancer cells to evaluate their interaction and immune cell infiltration to provide an innovative in vitro screening of immunomodulatory agents and biomarkers. Methods 3D tumor microtissues were cultivated using a hanging drops system. Human non-small-cell lung cancer cell lines were incubated for 7 days to form microtissues. On day 5, peripheral blood mononuclear cells (PBMC) were added with or without interleukin-2 (IL-2) for 24 or 48 h. Viability of cancer cells and the infiltrating PBMC subpopulations were investigated by flow cytometry. Aggregation of tumor cells and PBMC and the infiltration of the PBMC into the tumor microtissues were analyzed by immunohistochemistry. Quantification of infiltration was measured by applying the TissueFAXS system. Results Immunohistochemistry revealed PBMC infiltration in all cell lines which increased under IL-2 stimulation. Analysis of infiltrating populations showed both lymphocyte subpopulations and monocytes within the tumor microtissues. In all three co-cultures, CD3+CD8+ and CD3+CD8+CD45R0+CD28+ lymphocytes were increased with IL-2, whereas CD3+CD8+CD45R0-CD28+ PBMCs were decreased with and without IL-2 stimulation. Conclusion In summary, we present a novel cell culture system to study the interaction between cancer cells and immune cells in 3-dimensional microtissues. In addition, we report for the first time an in vitro infiltration assay based on 3D microtissues. This model has the potential to provide a tool for ex-vivo drug testing and biomarker screening of immunomodulatory agents. [ABSTRACT FROM AUTHOR]

The epithelial-to-mesenchymal transition (EMT) is highly involved in the development of metastases. EMT transforms epithelial carcinoma cells into mesenchymal-like cells, characterized by increased cell migration and invasiveness. Transforming growth factor β (TGFβ) appears to be crucial in this process. Metformin and salinomycin have demonstrated an EMT inhibitory effect. The current experiments indicate that these substances specifically inhibit TGFβ-induced EMT in non-small cell lung cancer (NSCLC) cell lines. The NSCLC cell lines A549 and HCC4006 were stimulated with TGFβ for 48 h to induce EMT. Metformin or salinomycin was added simultaneously with TGFβ to inhibit TGFβ-induced EMT. Western blot analyses of E-cadherin and vimentin were performed to detect changes in EMT marker expression, and a wound healing assay was conducted to determine the potential effects on cell migration. The effects of the two drugs on cell viability were also investigated using MTS tetrazolium dye assays. The results revealed that cells undergoing EMT by application of TGFβ exhibited a downregulation of E-cadherin and an upregulation of vimentin protein expression on western blot analyses, and an increased capacity for cell migration. Simultaneous application of TGFβ and metformin specifically inhibited EMT and increased E-cadherin expression. At the higher dose tested, salinomycin also inhibited EMT, despite an increase in vimentin expression in the two cell lines. Furthermore, metformin and salinomycin, at the two concentrations tested, inhibited cell migration. These findings demonstrate that metformin and salinomycin are able to block EMT and inhibit EMT-induced cell migration. Thus, these two substances are novel EMT inhibiting drugs that have the potential to specifically control EMT and metastatic spread in NSCLC. [ABSTRACT FROM AUTHOR]

Both mast cells (MCs) and regulatory T cells (Tregs) have gained attention as immunosuppressive cell populations. To investigate a possible interaction, we used the Th1- and Th17-dependent model of nephrotoxic serum nephritis (NTS), in which both MCs and Tregs have been shown to play a protective role. Transfer of wild-type (wt) Tregs into wt recipients almost completely prevents development of NTS and leads to a profound increase of MCs in the renal draining lymph nodes (LNs). By contrast, transfer of wt Tregs into animals deficient in MCs, which are characterized by an exaggerated susceptibility to NTS, no longer exhibited protective effects. Blocking the pleiotropic cytokine IL-9, known to be involved in MC recruitment and proliferation, by means of a mAb in mice receiving Tregs abrogated protection from NTS. Moreover, transfer of IL-9-deficient Tregs also failed to protect from NTS. In the absence of Treg-derived IL-9, MCs fail to accumulate in the LNs, despite the fact that IL-9 deficiency does not alter the general suppressive activity of Tregs. In summary, to our knowledge, we provide the first direct in vivo evidence that the nephroprotective, anti-inflammatory effects of Tregs critically depend on IL-9-mediated attraction of MCs into kidney-draining LNs. [ABSTRACT FROM AUTHOR]

Polymorphonuclear neutrophils (PMNs) are the first line of defence against invading pathogens. They contain a multitude of antimicrobial and potentially cytotoxic substances packed in granules and secretory vesicles. Here, we show that granzyme A (GzmA) is constitutively expressed in human PMNs, but not in the promyelocytic cell line HL-60, by performing flow cytometry, western blot, enzyme-linked immunosorbent assay and quantitative polymerase chain reaction. To further track the intracellular localization of GzmA, we performed subcellular fractionation and found GzmA to be present in peroxidase-negative granules. Finally, stimulation with opsonized Escherichia coli or the bioincompatible haemodialysis membrane cuprophane led to up-regulation of GzmA expression at the transcriptional level as well as at the translational level. In conclusion, we provide clear evidence that GzmA is constitutively expressed in human PMNs and can be up-regulated upon stimulation. These findings may also indicate a role for GzmA in PMNs in defence against invading pathogens. [ABSTRACT FROM AUTHOR]

Objective: This study aimed to develop an observer-rated measure of Insight into Conflictual Relationship Patterns (ICR) applicable to audio- or videotapes of psychotherapy sessions and conduct a first psychometric evaluation. Method: We investigated the item properties, principal components, reliability, and validity of the ICR in a naturalistic sample of N = 125 outpatients in psychodynamic psychotherapy. Results: The ICR consists of 12 items that showed adequate item discrimination and item difficulty indices. All items represent one principal component. Using item response theory, discrimination parameters and item characteristic curves revealed that the ability of all items to differentiate patients was adequate to very good. The scale demonstrated good interrater reliability (ICC(3,1) =.76–.93), adequate internal consistency (Cronbach's α =.84), and high retest reliability (r =.91). Regarding validity, the ICR was significantly associated with insight according to the Achievement of Therapeutic Objectives Scale and patient-perceived session depth. Insight at session five predicted a symptomatic increase from session five to session ten. Conclusion: The ICR is an observer-rated measure to assess insight from psychotherapy session recordings that has demonstrated several aspects of reliability and validity. Future studies are needed to clarify the impact of ICR-assessed insight for symptomatic outcome. [ABSTRACT FROM AUTHOR]

Introduction: We describe a novel 3D co-culture model using non-small cell lung cancer (NSCLC) cell lines in combination with lung fibroblasts. This model allows the investigation of tumour-stroma interactions and addresses the importance of having a more in vivo like cell culture model. Methods: Automation-compatible multi-well hanging drop microtiter plates were used for the production of 3D mono- and co-cultures. In these hanging drops the two NSCLC cell lines A549 and Colo699 were cultivated either alone or co-cultured with lung fibroblasts. The viability of tumour spheroids was confirmed after five and ten days by using Annexin V/Propidium Iodide staining for flow-cytometry. Tumour fibroblast spheroid formation was characterized by scanning electron microscope (SEM), semi-thin sections, fluorescence microscope and immunohistochemistry (IHC). In addition to conventional histology, protein expression of E-Cadherin, vimentin, Ki67, fibronectin, cytokeratin 7 and α-smooth muscle actin (α-SMA) was investigated by IHC. Results: Lower viability was observed in A549 monocultures compared to co-cultures, whereas Colo699 monocultures showed better viability compared to co-cultures. Ki67 expression varied significantly between mono- and co-cultures in both tumour cell lines. An increase of vimentin and decreased E-Cadherin expression could be detected during the course of the cultivation suggesting a transition to a more mesenchymal phenotype. Furthermore, the fibroblast cell line showed an expression of α-SMA only in co-culture with the cancer cell line A549, thereby indicating a mesenchymal to mesenchymal shift to an even more myofibroblast phenotype. Conclusion: We demonstrate that our method is a promising tool for the generation of tumour spheroid co-cultures. Furthermore, these spheroids allow the investigation of tumour-stroma interactions and a better reflection of in vivo conditions of cancer cells in their microenvironment. Our method holds potential to contribute to the development of anti-cancer agents and support the search for biomarkers. [ABSTRACT FROM AUTHOR]

We describe a sequential multistaining protocol for immunohistochemistry, immunofluorescence and CyTOF imaging for formalin-fixed, paraffin-embedded specimens (FFPE) in the formalin gas-phase (FOLGAS), enabling sequential multistaining, independent from the primary and secondary antibodies and retrieval. Histomorphologic details are preserved, and crossreactivity and loss of signal intensity are not detectable. Combined with a DAB-based hydrophobic masking of metal-labeled primary antibodies, FOLGAS allows the extended use of CyTOF imaging in FFPE sections. [ABSTRACT FROM AUTHOR]

Lipocalin-2 (Lcn-2) is involved in divergent processes such as acute kidney injury or bacterial host defence. Our study was designed to evaluate the functional role of Lcn-2 in nephrotoxic serum nephritis (NTS). Since Lcn-2 is expressed in tubular epithelial cells as well as in cells of innate immunity such as macrophages and polymorphonuclear neutrophils (PMN), we induced NTS in wild-type (WT), Lcn-2 knock-out (KO) mice and WT/Lcn-2 KO chimeras. Mice lacking Lcn-2 exhibited more glomerular damage with increased proteinuria and interstitial leukocyte accumulation compared to WT mice. Chimeras able to express Lcn-2 in macrophages and PMN but not in epithelial cells were found to develop NTS comparable to wild-type controls. In contrast, chimeras expressing Lcn-2 in tubular epithelial cells with no expression in innate immune cells developed increased NTS due to decreased concerted apoptosis but increased necrosis and formation of damage-associated molecular patterns (DAMPs) such as high-mobility group box 1 (HMGB-1) in the kidney. In vivo blockade of HMGB-1, a toll-like receptor (TLR)-2 agonist, significantly reduced inflammation and NTS in Lcn-2 knock-out mice. In parallel, TLR-2 signalling was found to drive Lcn-2 transcription in vitro. Taken together, Lcn-2 expressed in innate immune cells is protective in NTS by inducing concerted apoptosis and inhibiting the formation of HMGB-1 thereby limiting cytokine production via TLR-2 signalling. In parallel, TLR-2 dependent transcription of Lcn-2 is an endogenous inhibitor of inflammation in NTS. [ABSTRACT FROM AUTHOR]

Background: Liver-selective thyromimetics have been reported to efficiently reduce plasma cholesterol through the hepatic induction of both, the low-density lipoprotein receptor (LDLr) and the high-density lipoprotein (HDL) receptor; the scavenger receptor class B type I (SR-BI). Here, we investigated the effect of the thyromimetic T-0681 on reverse cholesterol transport (RCT) and atherosclerosis, and studied the underlying mechanisms using different mouse models, including mice lacking LDLr, SR-BI, and apoE, as well as CETP transgenic mice. Methodology/Principal Findings: T-0681 treatment promoted bile acid production and biliary sterol secretion consistently in the majority of the studied mouse models, which was associated with a marked reduction of plasma cholesterol. Using an assay of macrophage RCT in mice, we found T-0681 to significantly increase fecal excretion of macrophage-derived neutral and acidic sterols. No positive effect on RCT was found in CETP transgenic mice, most likely due to the observed decrease in plasma CETP mass. Studies in SR-BI KO and LDLr KO mice suggested hepatic LDLr to be necessary for the action of T-0681 on lipid metabolism, as the compound did not have any influence on plasma cholesterol levels in mice lacking this receptor. Finally, prolonged treatment with T-0681 reduced the development of atherosclerosis by 60% in apoE KOs on Western type diet. In contrast, at an earlier time-point T-0681 slightly increased small fatty streak lesions, in part due to an impaired macrophage cholesterol efflux capacity, when compared to controls. Conclusions/Significance: The present results show that liver-selective thyromimetics can promote RCT and that such compounds may protect from atherosclerosis partly through induction of bile acid metabolism and biliary sterol secretion. On-going clinical trials will show whether selective thyromimetics do prevent atherosclerosis also in humans. [ABSTRACT FROM AUTHOR]

Abstract: Background: We recently showed that aspirin promotes scavenger receptor class-B type I (SR-BI) protein expression in vitro in primary human macrophages and in vivo in resident peritoneal macrophages of mice. Methods: We compared SR-BI and CD68 expression in carotid atherosclerotic specimens from endarterectomized patients with (n =38) or without (n =19) low-dose aspirin medication (100mg/day) prior to endarterectomy. Results: We found no differences concerning expression of CD68, indicating that aspirin did not influence macrophage content within atherosclerotic plaques. However, aspirin increased the expression of SR-BI protein in the analyzed specimens. In human THP-1-derived macrophages, induction of SR-BI protein by aspirin was abrogated by concomitant pharmacological inhibition of nuclear factor-kappa B (NF-κB). In in vitro experiments employing cultured primary macrophages from NF-κB/p50 KO mice, aspirin was not able to influence SR-BI expression. Additionally, no considerable effects on SR-BI expression were observed in vivo in resident macrophages of NF-κB/p50 KO mice orally treated with low or high doses of aspirin, respectively. Conclusions: We suggest that aspirin treatment might lead to enhanced expression of SR-BI in human plaque macrophages and that this effect is dependent on the presence of NF-κB. [Copyright &y& Elsevier]

In many clinical settings, the duration of renal ischemia and therefore the outcome of acute renal failure cannot be determined adequately. Renal ischemia reperfusion injury is known to shorten telomeres and upregulate stress-induced genes, such as the cyclin-dependent kinase (CDK) inhibitor p21. So far, the expression and role of CDK inhibitors, as well as mouse telomerase reverse transcriptase (mTERT), has not been investigated in a model with variable lasting ischemic periods. Male C57B1/6 mice were subjected to renal ischemia reperfusion injury by clamping both renal pedicles for 10, 20, 30, and 45 min, and the kidneys were allowed to be reperfused for 3, 24, and 48 h. Expression of different CDK inhibitors and mTERT was evaluated. Mice developed signs of acute renal failure linear to the duration of the ischemic period. Real-time PCR revealed that mTERT was only significantly upregulated in kidneys after short ischemic periods (20 mm). In contrast, p21 was constantly upregulated in kidneys after long ischemic intervals (30 and 45 mm), but not in kidneys, which were clamped for shorter periods. Mainly, tubular cells contributed to the observed increase in p21 expression. Targeting p21 via the selective p53 inhibitor pifithrin-α was able to prevent acute renal failure when administered immediately before ischemia. The expression of another CDK inhibitor, namely p16, was differentially regulated, depending on the time of reperfusion. Taken together, we detected mTERT and p21 as "indicator" genes for short and long ischemic intervals, respectively. These two proteins might also be possible new therapeutic targets in the treatment and prevention of acute renal failure. [ABSTRACT FROM AUTHOR]