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The primary consequence of positive selection is to render thymocytes responsive to cytokines and chemokines expressed in the thymic medulla. In the present study, our main objective was to discover which cytokines could support the differentiation of positively selected thymocytes. To this end, we have developed an in vitro model suitable for high-throughput analyses of positive selection and CD8 T-cell differentiation. The model involves coculture of TCRhiCD5intCD69− double-positive (DP) thymocytes with peptide-pulsed OP9 cells and γc-cytokines. We report that IL-4, IL-7, and IL-21 have nonredundant effects on positively selected DP thymocytes. IL-7 signaling phosphorylates STAT5 and ERK; induces Foxo1, Klf2, and S1pr1; and supports the differentiation of classic CD8 T cells. IL-4 activates STAT6 and ERK and supports the differentiation of CD8intPD-L1hiCD44hiEOMES+ innate CD8 T cells. IL-21 is produced by thymic epithelial cells and the IL-21 receptor-α is strongly induced on DP thymocytes undergoing positive selection. IL-21 signaling phosphorylates STAT3 and STAT5, but not ERK, and does not support CD8 T-cell differentiation. However, IL-21 has a unique ability to up-regulate BCL-6, expand DP thymocytes undergoing positive selection, and increase the production of mature T cells. Our data suggest that injection of recombinant IL-21 might enhance thymic output in subjects with age- or disease-related thymic atrophy.

Based on transcriptomic analyses of thousands of samples from The Cancer Genome Atlas, we report that expression of constitutive proteasome (CP) genes (PSMB5, PSMB6, PSMB7) and immunoproteasome (IP) genes (PSMB8, PSMB9, PSMB10) is increased in most cancer types. In breast cancer, expression of IP genes was determined by the abundance of tumor infiltrating lymphocytes and high expression of IP genes was associated with longer survival. In contrast, IP upregulation in acute myeloid leukemia (AML) was a cell-intrinsic feature that was not associated with longer survival. Expression of IP genes in AML was IFN-independent, correlated with the methylation status of IP genes, and was particularly high in AML with an M5 phenotype and/or MLL rearrangement. Notably, PSMB8 inhibition led to accumulation of polyubiquitinated proteins and cell death in IPhigh but not IPlow AML cells. Co-clustering analysis revealed that genes correlated with IP subunits in non-M5 AMLs were primarily implicated in immune processes. However, in M5 AML, IP genes were primarily co-regulated with genes involved in cell metabolism and proliferation, mitochondrial activity and stress responses. We conclude that M5 AML cells can upregulate IP genes in a cell-intrinsic manner in order to resist cell stress.

Purpose To describe the outcomes of peer review across all 14 cancer centers in Ontario. Methods and Materials We identified all peer-reviewed, curative treatment plans delivered in Ontario within a 3-month study period from 2013 to 2014 using a provincial cancer treatment database and collected additional data on the peer-review outcomes. Results Considerable variation was found in the proportion of peer-reviewed plans across the centers (average 70.2%, range 40.8%-99.2%). During the study period, 5561 curative plans underwent peer review. Of those, 184 plans (3.3%) had changes recommended. Of the 184 plans, the changes were major (defined as requiring repeat planning or having a major effect on planning or clinical outcomes, or both) in 40.2% and minor in 47.8%. For the remaining 12.0%, data were missing. The proportions of recommended changes varied among disease sites (0.0%-7.0%). The disease sites with the most recommended changes to treatment plans after peer review and with the greatest potential for benefit were the esophagus (7.0%), uterus (6.7%), upper limb (6.3%), cervix and lower limb (both 6.0%), head and neck and bilateral lung (both 5.9%), right supraclavicular lymph nodes (5.7%), rectum (5.3%), and spine (5.0%). Although the heart is an organ at risk in left-sided breast treatment plans, the proportions of recommended changes did not significantly differ between the left breast treatment plans (3.0%, 95% confidence interval 2.0%-4.5%) and right breast treatment plans (2.4%, 95% confidence interval 1.5%-3.8%). The recommended changes were more frequently made when peer review occurred before radiation therapy (3.8%) than during treatment (1.4%-2.8%; P =.0048). The proportion of plans with recommended changes was not significantly associated with patient volume ( P =.23), peer-review performance ( P =.36), or center academic status ( P =.75). Conclusions Peer review of treatment plans directly affects the quality of care by identifying important clinical and planning changes. Provincial strategies are underway to optimize its conduct in radiation oncology.

The majority of newly diagnosed prostate cancers are slow growing, with a long natural life history. Yet a subset can metastasize with lethal consequences. We reconstructed the phylogenies of 293 localized prostate tumors linked to clinical outcome data. Multiple subclones were detected in 59% of patients, and specific subclonal architectures associate with adverse clinicopathological features. Early tumor development is characterized by point mutations and deletions followed by later subclonal amplifications and changes in trinucleotide mutational signatures. Specific genes are selectively mutated prior to or following subclonal diversification, including MTOR, NKX3-1, and RB1. Patients with low-risk monoclonal tumors rarely relapse after primary therapy (7%), while those with high-risk polyclonal tumors frequently do (61%). The presence of multiple subclones in an index biopsy may be necessary, but not sufficient, for relapse of localized prostate cancer, suggesting that evolution-aware biomarkers should be studied in prospective studies of low-risk tumors suitable for active surveillance.

Background The optimal transfusion threshold after surgery in children is unknown. We analyzed the general surgery subgroup of the TRIPICU (Transfusion Requirements in Pediatric Intensive Care Units) study to determine the impact of a restrictive versus a liberal transfusion strategy on new or progressive multiple organ dysfunction syndrome (MODS). Methods The TRIPICU study, a prospective randomized controlled trial conducted in 17 centers, enrolled a total of 648 critically ill children with a hemoglobin equal to or below 9.5 g/dL within 7 days of pediatric intensive care unit (PICU) admission to receive prestorage leukocyte-reduced red-cell transfusion if their hemoglobin dropped below either 7.0 g/dL (restrictive) or 9.5 g/dL (liberal). A subgroup of 124 postoperative patients (60 randomized to restrictive and 64 to the liberal group) were analyzed. This study was registered at http://www.controlled-trials.com and carries the following ID ISRCTN37246456. Results Participants in the restrictive and liberal groups were similar at randomization in age (restrictive vs. liberal: 53.5 +/- 51.8 vs. 73.7 +/- 61.8 months), severity of illness (pediatric risk of mortality [PRISM] score: 3.5 +/- 4.0 vs. 4.4 +/- 4.0), MODS (35% vs. 29%), need for mechanical ventilation (77% vs. 74%), and hemoglobin level (7.7 +/- 1.1 vs. 7.9 +/- 1.0 g/dL). The mean hemoglobin level remained 2.3 g/dL lower in the restrictive group after randomization. No significant differences were found for new or progressive MODS (8% vs. 9%; P = 0.83) or for 28-day mortality (2% vs. 2%; P = 0.96) in the restrictive versus liberal group. However, there was a statistically significant difference between groups for PICU length of stay (7.7 +/- 6.6 days for the restrictive group vs. 11.6 +/- 10.2 days for the liberal group; P = 0.03). Conclusions In this subgroup analysis of pediatric general surgery patients, we found no conclusive evidence that a restrictive red-cell transfusion strategy, as compared with a liberal one, increased the rate of new or progressive MODS or mortality.

Patient-reported outcomes (PROs) report patients’ assessments of the impact of a health condition and its treatment, and can promote patient-centered care. Objectives To address the effectiveness of graphic display of PRO data in clinical practice by reviewing existing literature, and current recommendations, regarding graphic presentations of PROs. Methods We performed an integrated literature review to identify themes and emerging principles guiding effective graphic display of PRO data. The findings were placed in the context of the literature informing graphical presentation of other clinical data. Results Although a large body of literature informs graphical presentation of clinical data, only nine empirical studies addressed presentation of PROs. Four major themes emerged: many patients and most clinicians can accurately interpret some PRO graphs; interpretation accuracy, personal preference, and perceived level of understanding are sometimes discordant; patient age and education may predict PRO graph comprehension; patients tend to prefer simpler graphs than do clinicians. Conclusions Little empirical research specifically addresses graphic representation of PRO data. A single format may not work optimally for both clinicians and patients. Practice implications Patients and clinicians may or may not comprehend PRO data when graphically presented. Further research to determine best practices for presenting PROs optimally is needed.

Because of the often episodic nature of wastewater toxicity, routine monitoring using expensive and time consuming tests can constitute an inefficient means of toxicity evaluation, particularly when negative results are generated. Cost-effective screening tests enabling the rapid detection of effluent toxicity are clearly needed, and they should be used to rapidly determine where in-depth investigations should be focused. The LuminoTox is a recently-developed screening test enabling the rapid determination of wastewater toxicity. This test is based on the inhibition of chlorophyll fluorescence emitted by photosynthetic systems. The combined use of photosynthetic enzyme complexes (PECs), isolated from higher plants, and whole photosynthetic organisms (algae) allows a wide range of toxic inhibitors to be detected within 10-15 min. The detection thresholds obtained for individual toxic chemicals indicate that algae are less sensitive to metal cations than PECs, because of the algal cell wall being ion selective. However, other toxic chemicals, such as phenolic compounds and nitrogen ammonia, acting on the last constituents of the photosynthetic enzyme complex that are degraded during the PEC extraction process, are more easily detected with algae after just 10 min of exposure. The combination of PECs and algae is not only useful for rapid toxicity screening, but yields results that are as sensitive as those of standard bioassays. Toxicity data generated with mining industry effluents demonstrate that PECs routinely prove to be as sensitive as daphnia, while algal sensitivity is comparable to that of the standard trout bioassay. An important feature of LuminoTox and algal photosynthetic system testing, however, resides in the production of their rapid and sensitive responses (10-15 min) in comparison with those of the more traditional tests (48-96 h for daphnia and trout, respectively).

Shellfish have the capacity to accumulate chemical contaminants found in their biotope and therefore present a potential risk for consumers. This study was conducted to assess the chemical risks associated with consumption of shellfish harvested on the north shore of the St. Lawrence River's lower estuary. A survey was carried out on 162 recreational harvesters, and shellfish were sampled for chemical contaminant analysis. We quantified 10 metals, 22 polycyclic aromatic hydrocarbons (PAHs), 14 polychlorinated biphenyls (PCBs), and 10 chlorinated pesticides. We subsequently evaluated cancer and noncancer risks for four consumption scenarios based on our survey results and published results. Soft-shell clams (Mya arenaria) were by far the most consumed shellfish species. Of the 56 selected contaminants, 36 were detected in the 23 homogenates of soft-shell clam meat. None of the contaminants found in the soft-shell clams were associated with intakes that exceed the main exposure limit recommendations proposed to prevent noncancer effects. However, several limits must be considered before drawing conclusions about the relative safety of shellfish consumption regarding this end point. Furthermore, inorganic arsenic and PCBs were present in sufficient concentrations to lead to cancer risks exceeding the level often considered acceptable for environmental exposure (1 x 10 (-4) to 1 x 10(-6)) in each of the four scenarios, even for the lowest observed scenario of 15 meals of soft-shell clams per year.

Psmb11 is a catalytic subunit of the thymoproteasome, expressed exclusively in cTECs (cortical thymic epithelial cells). It has been shown that Psmb11-mediated positive selection is pivotal for the generation of an immunocompetent T-cell repertoire. We found that lack of Psmb11 results in a suite of gene expression changes in cTECs and not in mTECs, about 200 genes with enrichment in cell-cell interaction and antigen processing and presentation. One yet unsolved question is why the cellularity of CD8+ single-positive (SP8) but not SP4 thymocytes decreases strongly as a consequence of Psmb11-deficiency. Our transcriptomic analyses of pure SM, M1 and M2 subsets of developing SP4 and SP8 T cells show that the maturation of SP4 cells is essentially independent of Psmb11. However SP8 thymocytes present a dysregulated development with impaired migration, survival and lineage commitment in Psmb11 knock-out mice, also detected at the protein level by flow-cytometry. Thymocytes present decreased surface levels of CD127 on SP8 and slightly increased CD5 on all subsets. This indicates a stronger activation of the developing T cells, which alone cannot explain the impact on the SP8 cellularity. We have identified a novel role of Psmb11 in modulating, directly or indirectly, the expression of genes in cTECs, which is necessary to guide and complete the positive selection of SP8 thymocytes.

Although health-related quality of life (HRQL) is increasingly assessed in randomized controlled trials (RCTs), HRQL findings are not always incorporated into clinical decision making. We examined the current perspectives of oncologists on the use of HRQL findings from RCTs, and variation of these perspectives between countries and specialties. A cross-sectional e-survey of oncologist members of the NCIC Clinical Trials Group, the United Kingdom National Cancer Research Institute Clinical Studies Groups, and the Australia/New Zealand cancer clinical trials network was conducted. Respondents reported their perceptions of the usefulness of RCT HRQL outcomes for informing practice, their use of HRQL findings in practice, barriers/facilitators to this use, and preferences for HRQL data presentation in RCT publications. Chi-square tests compared responses between countries and specialties. A total of 396 oncologists participated (estimated response rate: 30 %). The most frequently reported specialties were medical (46 %) and radiation (37 %) oncology. HRQL findings from RCTs were reported as useful (73 %), but were infrequently used to guide clinical decisions with patients. Perceived barriers were lack of time (67 %) and understanding (57 %), and concerns about generalizability of results (68 %). Identified facilitators included joint publication of HRQL/clinical outcomes (96 %) and summary of clinical implications of HRQL findings in RCT publications (76 %). Use of HRQL findings, perceived barriers/facilitators, and presentation preferences did not differ by country or specialty. Oncologists support HRQL findings from RCTs, but perceive important barriers to their use in clinical decision making, regardless of country or specialty. Combined, clear reporting of HRQL/clinical data may facilitate their clinical application.

Kathleen Canning, Languages of Labor and Gender: Fernale Factory Work in Germany, 1850-1914, New York 1996., Moving the Social, Vol 20 (1998): Forschungen und Forschungsberichte

Historians have generally interpreted the early years of the Weimar Republic as an important stage in the development of the German welfare state. For the first time in the history of Germany, the state established in the constitution not only its own wideranging responsibilities and opportunities for intervention, but also the political and social rights of its citizens. Apart from “fundamentally” equal citizenship rights for womenand men (Art. 108) these also included entitlement to state support for the family and maternity as well as special state protection for marriage which, the constitution proclaimed, was to rest on an “equality of the two sexes” (Art. 119).

Both physiological and psychological stress cause thymic atrophy via glucocorticoïd (GC)-dependent apoptosis of double-positive (DP) thymocytes. Given the pervasiveness of stress, GC-induced thymic atrophy is arguably the most common type of acquired immunodeficiency. We recently reported that interleukin-21 (IL-21) has a unique ability to expand the small subset of DP thymocytes (CD69(+)) which are ongoing positive selection, and that administration of IL-21 increases thymic output in aged mice. The goal of this study was to evaluate whether IL-21 could mitigate GC-induced thymic atrophy. In contrast to double-negative (DN) and single-positive (SP) thymocytes, most DP thymocytes (CD69(-)) do not constitutively express the IL-21 receptor (IL-21R). Accordingly, CD69(-) DP thymocytes from PBS-treated mice were unresponsive to IL-21 administration. However, following GC injection, surviving CD69(-) DP thymocytes up-regulated IL-21R and responded to IL-21 treatment as evidenced by enhancement of Bcl6 expression and phosphorylation of STAT1, STAT3 and STAT5. Consequently, IL-21 administration to GC-treated mice accelerated thymic recovery by expanding considerably DP thymocytes and, to a lesser extent, DN thymocytes. However, IL-21-induced expansion of DN/DP thymocytes did not alter the diversity of the intrathymic or peripheral T-cell receptor (TCR) repertoire. We conclude that IL-21 dramatically accelerates recovery from GC-induced thymic atrophy.

205 Background: Review of treatment plans by a second radiation oncologist is an important quality indicator in radiation oncology. Peer review (PR) can improve quality of care in individual patients by detecting clinical and planning issues and recommending plan changes. This study reports the frequency and nature of these changes across all 14 radiation oncology programs (ROPs) in Ontario, Canada. Methods: We identified all peer-reviewed curative treatment plans delivered in Ontario within a 3-month study period between Dec 2013-Nov 2014 using Cancer Care Ontario’s Activity Level Reporting System, where data on treatment intent and date, disease site treated, PR status, timing of PR, and nature of recommended changes were available. Results: There was considerable variation in the proportion of plans peer-reviewed across ROPs (70.2%, range: 40.8-99.2%). Over the study period, 5,561 curative treatment plans were peer-reviewed and 3.3% had changes recommended. Of those, 21.0% had major clinical and re-planning implications. Recommended changes most often involved minor (63.1%) vs major (36.9%) re-planning implications. Highest proportions of changes were recommended for the treatment of the esophagus, uterus, upper limb, cervix, lower limb, H&N, bilateral lung, right supraclavicular nodes, rectum, and spine (5.0%-7.0%). Plans involving the left breast had slightly more changes recommended (3.0% [95%CI:2.0%-4.5%]) vs right breast (2.4% [95%CI:1.5%-3.8%]). Recommendations were more frequently made when PR was conducted pre-radiotherapy (3.8%) vs during (1.4%-2.8%; p = 0.005), however the nature and implementation of changes were not statistically associated with the timing of PR (p = 0.91; p = 0.23, respectively). Proportion of recommended changes to treatment plans was not statistically associated with ROP patient volume (p = 0.08), proportion of plans peer-reviewed (p = 0.36) or academic status (p = 0.75). Conclusions: Significant variation exists in the proportion of recommended changes across all disease sites and ROPs. PR seems effective in detecting treatment plans with important clinical and planning issues; strategies should be developed to optimize its conduct in radiation oncology.

Human carcinomas often show resistance to cisplatin and Bcl-2 is associated with resistance to cisplatin. However, Bcl-2 regulation on cisplatin treatment in human cancers is unknown. Here, we show a novel mechanism by which cisplatin treatment promotes resistance by increasing the expression of Bcl-2 mRNA. Bcl-2 mRNA and protein expression was increased in cisplatin-resistant endometrial cancer cell lines (KLE and HEC-1-A), but not in cisplatin-sensitive cell line (Ishikawa). Cisplatin-mediated increase in Bcl-2 expression was blocked by combination with either actinomycin D or cycloheximide. In addition, Bcl-2 inhibition by HA14-1 led to increased cisplatin-induced apoptosis in KLE and HEC-1-A, whereas Bcl-2 overexpression in Ishikawa led to decreased cisplatin-induced apoptosis. Inhibition of protein kinase C (PKC) activity prevented cisplatin-dependant increase in Bcl-2 mRNA, and induced apoptosis in KLE cells. Furthermore, PKC inhibition was associated with decreased Akt and NF-κB activity. Cells stably expressing shRNA for Akt isoforms revealed that Akt2 was involved in cisplatin-dependant increase in Bcl-2 and apoptosis. Overexpression of Akt2 in Akt2-deficient cells led to increased Bcl-2 expression on cisplatin treatment. Our data suggest a novel regulation pathway of Bcl-2 by cisplatin, via the activation of PKC and Akt2, which has a profound impact on resistance to cisplatin-induced apoptosis in endometrial cancer cells.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Babesia microti is a tick-borne red blood cell parasite that causes babesiosis in people. Its most common vertebrate reservoir is the white-footed mouse. To determine whether B. microti invades reticulocytes, as does the canine pathogen B. gibsoni , we infected the susceptible inbred mouse strains C.B-17.scid and DBA/2 with a clinical isolate of B. microti . Staining of fixed permeabilized red blood cells with 4′,6′-diamidino-2-phenylindole or YOYO-1, a sensitive nucleic acid stain, revealed parasite nuclei as large bright dots. Flow cytometric analysis indicated that parasite DNA is primarily found in mature erythrocytes that expressed Babesia antigens but not the transferrin receptor CD71. In contrast, CD71-positive reticulocytes rarely contained Babesia nuclei and failed to express Babesia antigens. Accordingly, the frequency of YOYO-1-positive, CD71-negative cells strongly correlated with parasitemia, defined as the frequency of infected red blood cells assessed on Giemsa-stained blood smears. Importantly, the absolute numbers generated by the two techniques were similar. Parasitemia was modest and transient in DBA/2 mice but intense and sustained in C.B-17.scid mice. In both strains, parasitemia preceded reticulocytosis, but reticulocytes remained refractory to B. microti . In immunocompetent C.B-17 mice, reticulocytosis developed early, despite a marginal and short-lived parasitemia. Likewise, an early reticulocytosis developed in resistant BALB/cBy and B10.D2 mice. These studies establish that B. microti has a tropism for mature erythrocytes. Although reticulocytes are rarely infected, the delayed reticulocytosis in susceptible strains may result from parasite or host activities to limit renewal of the mature erythrocyte pool, thereby preventing an overwhelming parasitemia.