Search

Medulloblastoma (MB) is the most common malignant pediatric brain tumor. MB is classified into four primary molecular subgroups: wingless (WNT), sonic hedgehog (SHH), Group 3 (G3), and Group 4 (G4), and further genomic and proteomic subtypes have been reported. Subgroup heterogeneity and few actionable mutations have hindered the development of targeted therapies, especially for G3 MB, which has a particularly poor prognosis. To identify novel therapeutic targets for MB, we performed mass spectrometry-based deep expression proteomics and phosphoproteomics in 20 orthotopic patient-derived xenograft (PDX) models of MB comprising SHH, G3, and G4 subgroups. We found that the proteomic profiles of MB PDX tumors are closely aligned with those of primary human MB tumors illustrating the utility of PDX models. SHH PDXs were enriched for NFκB and p38 MAPK signaling, while G3 PDXs were characterized by MYC activity. Additionally, we found a significant association between actinomycin D sensitivity and increased abundance of MYC and MYC target genes. Our results highlight several candidate pathways that may serve as targets for new MB therapies. Mass spectrometry data are available via ProteomeXchange with identifier PXD035070.

Objectives Patients with MM continue to relapse on current therapies, stressing the need for new immunotherapy targets. GPRC5D is an orphan receptor that is expressed on malignant plasma cells in MM. Talquetamab (JNJ-64407564) is a bispecific IgG4 antibody that binds to GPRC5D and CD3, redirecting T cell killing to MM cells. Updated phase 1 results of talquetamab at the recommended phase 2 dose (RP2D) in patients with RRMM are presented. Material and methods Eligible patients had RRMM or were intolerant to standard therapies. Patients received talquetamab intravenously (IV; range 0.5–180 μg/kg) or subcutaneously (SC; range 5.0–800 μg/kg) weekly or biweekly. The primary objectives of the study were identification of the RP2D in part 1 and talquetamab safety and tolerability at the RP2D in part 2. Adverse events (AEs) were graded by CTCAE v4.03; cytokine release syndrome (CRS) was graded per Lee 2014 criteria. Response was assessed per IMWG criteria. Results As of Feb 8, 2021, 174 patients had received talquetamab, 102 by IV and 72 by SC. Across parts 1 and 2 of the study, 28 patients were treated at the RP2D of weekly SC 405 μg/kg, with 10.0 and 60.0 μg/kg step-up doses. Patients treated at the RP2D had a median age of 61.5 years (range 46–80) and a median of 5.5 prior lines of therapy (range 2–14), with 100%/79% triple-class/penta-drug exposed; 71%/18% triple- class/penta-drug refractory; 86% refractory to last line of therapy; and 21% with prior B- cell maturation antigen–directed therapy. No dose-limiting toxicities at the RP2D occurred in part 1. At the RP2D, most common AEs were CRS (79%; grade 3 4%; median time to onset was day after SC injection), neutropenia (64%; grade 3/4 54%), anemia (57%; grade 3/4 29%) and dysgeusia (57%; all grade 1/2). Infections were reported in 32% of patients (grade 3/4 4%) and neurotoxicity in 7% (grade 3/4 none). 75% of patients dosed at the RP2D had skin-related AEs (grade 3/4 none), including 18% with nail disorders. In response-evaluable patients (n = 24), the overall response rate at the RP2D was 63%, with 50% ≥very good partial response; 9/17 (53%) evaluable triple-class refractory patients and 3/3 (100%) penta-drug refractory patients responded. Median time to first confirmed response at the RP2D was 1.0 mo (range 0.2–3.8); responses were durable and deepened over time (median follow-up 6.2 mo [range 2.7–9.7+] for responders at the RP2D). At the RP2D, exposure was maintained over the maximum EC90 target level and consistent T cell activation was observed. Discussion The promising efficacy, safety profile, and convenience of SC dosing support monotherapy development and combination approaches with talquetamab. Based on pharmacokinetic data, other SC dosing strategies are being explored. Conclusions Talquetamab showed a high clinical response rate and was well-tolerated in patients with RRMM treated at the RP2D of weekly 405 μg/kg SC.

BACKGROUND Extirpation of malignant tumors of the parotid results in creation of a complex facial defect often in combination with facial nerve palsy. This study presents the authors' experience using vastus lateralis muscle as a chimeric flap with anterolateral thigh flap to allow both soft-tissue reconstruction and dynamic reanimation in radical parotidectomy. METHODS A retrospective review of the medical records of cancer patients who had undergone radical parotidectomy and reconstruction using a chimeric vastus lateralis and anterolateral thigh flap between March of 2013 and May of 2017 was performed using the Sydney Head and Neck Cancer Institute database. The return of dynamic midface movement was the primary outcome investigated. Electronic, clinician-graded facial function scale grades were used to formally assess postoperative outcomes. RESULTS A total of 27 patients were included in the study with an average age of 72 years (range, 31 to 88 years). Thirteen patients (48 percent) had developed dynamic function by the end of the study period. Young age predicted a more rapid return to dynamic function (p = 0.018). Both being a woman and having an intact facial nerve before surgery improved dynamic midface movement (p = 0.005 and p = 0.036, respectively). On multivariable analysis, superior midface dynamic function was associated with neurotization using midface facial nerve branches as opposed to using nerve-to-masseter alone (p = 0.05). CONCLUSION The chimeric vastus lateralis and anterolateral thigh flap is a suitable option for restoring defects and dynamic function following radical parotidectomy. CLINICAL QUESTION/LEVEL OF EVIDENCE Therapeutic, IV.

Problem Sociodemographic identities, including race, culture, ethnicity, gender, and sexual orientation (race and culture), are recognized as important determinants of health, with significant impacts on patients' health outcomes, but teaching medical students about this is challenging. The authors sought to identify areas for improvement in delivery of critical content about race, culture, structural inequalities, and health disparities within a set of virtual patient cases used by U.S. medical schools and develop revision guidelines. Approach A workgroup (medical students and faculty) conducted a literature review in 2017 to identify challenges and best practices for teaching and learning about race and culture in medicine. Using an analytic framework informed by this review, they analyzed 63 Aquifer virtual patient teaching cases for effectiveness of the presentation of race and culture, resulting in six main themes describing common mistakes or pitfalls. They then developed an evidence-based guide for systematic case revision. Outcomes The authors present a novel, practical guide for medical educators to use to revise existing teaching cases and improve the delivery of critical concepts surrounding race and culture. This guide includes fundamental definitions and six sections to guide structured case revision based on the main themes. It includes examples of language, suggested edits, and the rationale and evidence for recommendations. Next steps Feedback from faculty and students regarding implementation of the guide and delivery of revised content in Aquifer cases will be critical in determining the guide's effectiveness. This structured guide may be adapted to a variety of teaching modalities in medicine.

Background: More than half of all cancer patients undergo radiation therapy (RT), which can be curative or palliative to improve cancer-specific outcomes and quality of life. However, radioresistance emerges and a cure is not always achieved. To date, researchers have studied T-cells and lymphocytes to understand response to RT, but little is known about myeloid cells. This diverse group of cells helps mount immunological responses, but can also be immunosuppressive by lessening the strength of T-cells. We hypothesized that our network-based analyses of single cell proteogenomic data generated from tumor-infiltrating immune cells can identify druggable proteins in pathways driving unwanted immunosuppressive changes, particularly among myeloid cells. Methods: Proteogenomic (CITE-Seq) analysis was conducted on CD45+ immune cells from unirradiated and irradiated orthotopic 4T1 murine mammary tumors three- and ten-days after tumor irradiation. Based on select proteins’ surface expression, CITE-Seq generated eight distinct clusters, including three myeloid cell clusters — monocytes, macrophages, and granulocytes. We used VIPER (virtual inference of protein activity by enriched regulon analysis), a machine learning-based algorithm that infers protein activities by taking regulatory gene expression networks and calculating the weighted expression on each protein’s targets, to create functional subclusters of the eight immune-cell populations. For subclusters enriched by RT, we used OncoTreat to identify active, druggable proteins. Results: Lymphocytes were depleted with radiation, especially ten days post-irradiation. While monocytes appeared homogeneous, one of three subclusters of macrophages appeared more resistant (i.e. enriched with radiation) and had 4 druggable targets (PSMA4, HSP90AB1, PARP1, APEX1) from OncoTreat. Similarly, three of eight subclusters of granulocytes appeared resistant, with two potential targets (PSMA4, S100A9). Conclusion: Suppressive populations were identified among the immune cells, specifically myeloid cells. We plan to experimentally test whether drugging the aforementioned targets in macrophages and granulocytes improves the overall efficacy of RT by augmenting an anti-tumor immune response. Affiliations: Aparna Krishnan1, Aleksandar Obradovic1, Catherine Spina2, Andrea Califano1,3,4,5,6 1 Department of Systems Biology* 2 Department of Radiation Oncology* 3 Herbert Irving Comprehensive Cancer Center* 4 Department of Medicine* 5 Department of Biochemistry & Molecular Biophysics* 6 Department of Biomedical Informatics* * Columbia University Irving Medical Center, New York, NY 10032, USA Citation Format: Aparna Krishnan, Aleksander Obradovic, Catherine Spina, Andrea Califano. Single cell network-based analyses defines the effect of radiation on the tumor immune microenvironment, including the response by and molecular determinants of suppressive myeloid cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6173.

Medulloblastoma (MB) is classified into four molecular subgroups: wingless (WNT), sonic hedgehog (SHH), Group 3 (G3) and Group 4 (G4), each with different molecular profiles and patient outcomes. Subgroup heterogeneity and low mutational burdens have hindered the identification of actionable therapeutic targets, especially in G3 MB which has a particularly poor prognosis. Therefore, we took a (phospho)-proteomics approach to identify active pathways and potential therapeutic opportunities in twenty orthotopic patient-derived xenograft (PDX) models of MB comprising SHH, G3 and G4 subtypes. Through our enrichment analysis, we identified processes and pathways specifically upregulated in each MB subgroup. We also utilized neural network derived kinase-substrate predictions and kinase activity scores inferred by a heuristic machine learning algorithm to further characterize phosphosignaling activity. We found that MB PDX models recapitulate many features of primary MB tumors including two distinct proteomic subtypes of G3. G3a was enriched for transcription, translation and MYC target genes while G3b was enriched for axon guidance and neurotrophin signaling pathways. Notably, both G3a and G3b contained higher abundance of mitochondrial proteins, suggesting altered tumor metabolism in G3 MB. SHH PDXs displayed increased NFκB and JNK-MAPK signaling. Group 4 MBs most closely resembled differentiated neuronal cells and were enriched for PKC and AMPK signaling as well as DNA repair pathways. In conclusion, we have provided a comprehensive proteomic and phosphoproteomic characterization of commonly studied MB PDX models and revealed new insights into subgroup enriched pathways and kinase activity in MB.

Medulloblastoma (MB), the most common malignant pediatric brain cancer, has four subtypes based on genomics profiling: Wingless (WNT), Sonic Hedgehog (SHH), Group 3, and Group 4. However, subtype heterogeneity and low mutation rates hinder the identification of actionable molecular targets within each group. Studies thus suggest a phosphoproteomics (study of protein phosphorylation), instead of a genomics approach, to better distinguish the MB subgroups. Thus, this research will measure MB’s irregular protein activity via protein phosphorylation patterns. Protein kinases phosphorylate proteins on specific amino acids, also known as substrates, to regulate their functions. Given that less than 5% of substrates have annotated kinase associations, it is difficult to identify the kinases in MB which irregularly phosphorylate their substrates in order to determine the most effective kinases for drugs to target. As a result, the publicly-available tool NetworKIN was employed to predict novel kinase-substrate interactions in this study to expand the currently limited annotated interactions. For a given dataset of substrates, NetworKIN will output kinasesubstrate predictions, each receiving a confidence score (higher score $=$ higher confidence) ranging from zero to the hundreds. To identify a score threshold for high confidence predictions, NetworKIN was run on the substrates in the Human PhosphoSite Plus Database (PSP), the largest curated database of experimentally verified kinase-substrate interactions in humans. Three statistical thresholding methods were tested and the optimal cutoff score was determined to lie at 5.88. This threshold was applied to the predictions for the MB dataset, resulting in 774 novel high confidence kinase-substrate interactions. To understand how these interactions altered the enriched kinases across the MB subgroups, kinase activity scores before and after adding the predictions were calculated for all 20 samples using the tool Inference of Kinase Activities from Phosphoproteomics. Subgroup classification based on these activity scores revealed that, with the addition of high-confidence interactions to PSP, there are two smaller subgroups of Group 3 and one of those subgroups more closely resembled the Group 4 samples. The alignment of these findings with current literature increases our confidence in the prediction algorithm and elucidates novel phosphorylation patterns to more accurately portray the phosphosignaling pathways driving MB tumorigenesis. Future cancer phosphoproteomics studies can also employ this robust prediction tool to continue expanding our understanding of aberrant kinase-substrate interactions in human diseases.

Background Skilled attendance at delivery is a key marker for reducing maternal mortality. Effective community engagement strategies complemented by community health worker (CHW) services can improve access to maternal health services in areas with limited health infrastructure or workforce. Methods A quasi-experimental study with matched comparison groups was conducted in Cambodia, Kenya and Zambia to determine the effect of integrated community investments on skilled birth attendance (SBA). In each country, communities in two districts/sub-districts received a package of community-oriented interventions comprised of timed CHW household health promotion for maternal, newborn and child health complemented by social accountability mechanisms using community scorecards. Two matched comparison districts/sub-districts received ongoing routine interventions. Data from the final evaluation were examined to determine the effect of timed CHW services and community-oriented interventions on SBA. Results Over 80% of the 3037 women in Cambodia, 2805 women in Kenya and 1171 women in Zambia reported SBA. Women in intervention sites who received timely CHW health promotion and social accountability mechanisms in Cambodia showed significantly higher odds of SBA (aOR = 7.48; 95% CI: 3.87, 14.5). The findings also indicated that women over the age of 24 in Cambodia, women with primary or secondary education in Cambodia and secondary education in Kenya, women from higher wealth quintiles in Cambodia, and women with four or more antenatal care (ANC) visits in all countries reported significantly higher odds of SBA. Inclusion of family members in pregnancy-related discussions in Kenya (aOR = 2.12; 95% CI: 1.06, 4.26) and Zambia (aOR = 6.78; 95% CI: 1.15, 13.9) and follow up CHW visits after a referral or health facility visit (aOR = 2.44; 95% CI: 1.30, 4.60 in Cambodia; aOR = 2.17; 95% CI 1.25, 3.75 in Kenya; aOR = 1.89; 95% CI: 1.05, 2.02 in Zambia) also showed significantly greater odds of SBA. Conclusions Enhancing people-centered care through culturally appropriate community-oriented strategies integrating timely CHW health promotion and social accountability mechanisms shows some evidence for improving SBA during delivery. These strategies can accelerate the achievement of the sustainable development goals for maternal child and newborn health.

Companies increasingly rely on virtual teams. Despite numerous studies examining the challenges of geographically dispersed work, the findings are often mixed. The purpose of this article is to ide...

In an era where mobile phones and computers are ubiquitous, technology-based interventions to reduce HIV and other sexually transmitted infections (STIs) have great potential to reach high-risk groups, including men who have sex with men (MSM). This study aimed to examine technology usage to find sexual health information online among MSM in Hanoi, Vietnam. A cross-sectional study of 205 MSM in Hanoi was conducted from February to May 2016. Overall, 50.7% of participants reported having used a smartphone, computer, or tablet to find HIV/STI testing locations in the past year, and 75.1% reported having used such devices to find other HIV/STI information online. Unemployment (adjusted prevalence ratio [aPR]: 1.13, 95%CI: 1.00–1.28) and having been tested for HIV (aPR: 1.27, 95%CI: 1.07–1.51) were significantly associated with using technology to find online sexual health information. MSM who had ever exchanged sex for money or drugs (aPR: 0.80; 95%CI: 0.68–0.94) were less likely to use technology to find sexual health information online. Technology is a promising platform for HIV/STI prevention programs among MSM, with the potential to reach different subgroups. Further efforts to develop technology-based interventions tailored to the needs of the MSM communities in Hanoi and to encourage MSM who were not currently seeking sexual health information and testing services online to do so are necessary.

Web service composition is a novel methodology to integrate diverse business solutions to accomplish complex business logic. In this fast-paced digital era with diverse categories of users, intelligent service composition based on the contextual information is highly essential to satisfy individual users requirements. For intelligent composition, ontology based declarative knowledge model is used to describe the business rules, commercial directions and personalized user environment in machine comprehensible way. This paper depicts healthcare service composition with domain ontology to encompass ontology manipulation, contextual information gathering, selecting services and dynamic composition. E-healthcare system automates service composition which invokes hospital, ambulance, scan-laboratory and pharmacy services. It provide valuable information like nearest hospital, best ambulance, scan-laboratory facilities, appropriate department and doctor details by gathering the contextual inforamtion of users like location, severity, symptoms, etc. Thus, elaborare search process is eliminated which ultimately helps the user to save time and life in case of emergency situations. thod.

Purpose – This paper aims to propose and empirically document the idea that people’s perceptions of having been treated fairly depend, in part, on whether the explanation provided to them of a product’s pricing is primarily based on the costs of labor (a service) versus materials (goods). Because materials are more fixed and tangible than the effort of labor, it is argued that people will have fewer counterfactual thoughts about how things could have been different with the cost of materials than those associated with labor. This has implications for fairness judgments more generally, as it suggests that people may be uneven in which types of data they attend to when making fairness judgments. Three experiments are presented that empirically test the relationship between the salience of goods versus services in the price paid and the resulting perceptions of fairness. Findings confirm that thoughts of money spent on a service were associated with lesser feelings of fairness than were thoughts of money spent on a good. This research uniquely identifies the mechanism by which some evaluations are considered fairer than others. Implications for organizational processes, such as procedural justice and fair compensation, are discussed. Design/methodology/approach – Three experiments are presented that empirically test the relationship between the salience of goods versus services in the price paid, and the resulting perceptions of fairness. Findings – Findings confirm that thoughts of money spent on a service were associated with lesser feelings of fairness than were thoughts of money spent on a good. Originality/value – This research uniquely identifies the mechanism by which some evaluations are considered fairer than others. Implications for organizational processes, such as procedural justice and fair compensation, are discussed.

As the number of microblogging tools and social media users rapidly increases, it is important to understand who and what is being disseminated. One emerging area of research is the use of Twitter as a means for the masses to voice dissent and mobilize action. This study analyzed the features of the top 30 tweeters and top 30 retweeted users during the 2011 protests in Egypt. By conducting a content analysis of tweets from the #jan25 data set (N = 600), we found that the characteristics of the source including location, media affiliation, longer account duration, and number of followers positively impacted retweetability. Power users or those tweeters sending the most messages were more likely to be based in Egypt than in other areas. Users who retweet messages seem to constitute a different group with more passive characteristics. The content of messages such as language intensity, information or URL sharing, and call to action were not significant predictors of retweetability. Because tweets had the sam...

Many men who have sex with men (MSM) in low and middle income countries search for male sexual partners via social media in part due to societal stigma and discrimination, yet little is known about the sexual risk profiles of MSM social media users. This cross-sectional study investigates the prevalence of social media use to find male sex partners in Hanoi, Vietnam and examines associations between social media use and sociodemographic and behavioral characteristics, including levels of internalized, perceived and enacted stigma, high-risk sexual behaviors, and HIV testing. 205 MSM were recruited from public venues where MSM congregate as well as through snowball sampling and completed an anonymous survey. MSM who found their male sexual partners using social media in the last year were more likely to have completed a university or higher degree (aOR 2.6; 95% CI 1.2-5.7), experience high levels of MSM-related perceived stigma (aOR 3.0; 95% CI 1.1-8.0), and have more than ten lifetime male sexual partners (aOR 3.2; 95% CI 1.3-7.6) compared to those who did not use social media. A niche for social media-based interventions integrating health and stigma-reduction strategies exists in HIV prevention programs for MSM.

Background Postictal delirium is a common adverse effect of electroconvulsive therapy (ECT) and can be dangerous to both patient and staff caring for them in the postanesthesia care unit. However, little is known about predictors of postictal delirium. Objectives The aim of this study was to identify predictors of postictal delirium. We hypothesized that both patient and ECT treatment variables might influence the likelihood of postictal delirium. Methods We prospectively monitored postictal delirium in the postanesthesia care unit using the Confusion Assessment Method for the Intensive Care Unit after the first ECT treatment of 96 consecutive patients. Patient and treatment variables were extracted retrospectively by chart review. A multiple logistic regression model was developed to assess the effect of these variables on the likelihood of developing delirium. Results Seizure length was found to be a statistically significant predictor of postictal delirium after adjusting for other covariates ( p = 0.003). No other variables were predictive. Conclusion A long ECT seizure increases the likelihood of delirium in the postanesthesia care unit at the first treatment. This finding suggests that postanesthesia care unit staff may benefit from knowledge about seizure length for predicting postictal delirium and anticipating the best management of post-ECT patients.

In this study, we have explored the use of mobile phones during negotiations. Specifically, we examined the effects that multitasking — reading messages on a mobile phone while negotiating face to face — had on the outcome achieved in a negotiation, as well as on perceptions of professionalism, trustworthiness, and satisfaction. Using an experimental design in a face-to-face dyadic negotiation, we found that multitasking negotiators achieved lower payoffs and were perceived as less professional and less trustworthy by their partners.

The United States faces a shortage of well-trained primary care physicians (PCPs). The demand for primary care services is projected to increase, with an estimated 50,000 more PCPs required by 2025...

Purpose The purpose of this exploratory study was to investigate independent school Boards as teams using a social cognitive perspective. Specifically, the study investigated Board processes and the nature of relationships between Board member self-efficacy, Board collective efficacy and performance of independent school Boards in New South Wales, Australia. Design/methodology/approach A multiple case study design that used qualitative research methods was employed. An expert steering group provided advice on the categorization of governance structures. A stratified purposeful sample of eight independent school Boards within the Sydney metropolitan area, New South Wales Australia participated. Data were collected from individual, semi-structured, face-to-face interviews with the Head of school, Board Chair and two Board members from each school. The interviews were digitally recorded and transcribed verbatim. Data were analyzed using qualitative data analysis procedures suggested in the literature. Findings The findings provide evidence that for independent school Board members in this study, self-efficacy and collective efficacy beliefs were related to perceptions of Board performance. Board member self-efficacy and Board collective efficacy appeared to be linked. Self-efficacy beliefs were primarily based on mastery experiences. Collective efficacy (at the individual level) primarily was based on members’ perceptions of Board past performance. Originality/value This paper provides insight into individual Board member beliefs likely to shape processes associated with independent school Board performance in New South Wales, Australia. The study is one of only a few that have adopted an empirical and descriptive approach, rather than only providing normative direction and imperatives.

Multidisciplinary teamwork is a key requirement in the design thinking approach to innovation. The tools currently available for effective team coaching are limited to heuristics derived from either experienced design thinking professionals or clinical psychology practitioners. Our research aims to improve this current situation by providing design thinking managers, coaches and instructors a scientifically validated tool for augmenting design team performance. We present the development of a software tool called the IDN Tool based on the Interaction Dynamics Notation to analyze team interactions and diagnose patterns of behavior that influence design outcomes. We demonstrate the use of the IDN Tool through analysis of the interaction behaviors of seven design teams engaged in a concept generation activity, which were independently rated by a two-person Jury using the criteria of utility and novelty. Through the analysis we were able to visually isolate the interaction behaviors that had a high positive or negative correlation with the levels of novelty and utility of concepts judged a priori. With further work, this has the potential of improving in-process design team performance with a positive influence on design outcomes.

The probability of survival is conventionally calculated from autologous hematopoietic cell transplantation (aHCT). Conditional survival takes into account the changing probability of survival with time survived, but this is not known for aHCT populations. We determined disease- and cause-specific conditional survival for 2388 patients treated with aHCT over a period of 20 years at a single institution. A total of 1054 deaths (44% of the cohort) were observed: 78% attributed to recurrent disease; 9% to subsequent malignancies and 6% to cardiopulmonary disease. Estimated probability of relative survival was 62% at 5 years and 50% at 10 years from aHCT. On the other hand, the 5-year relative survival was 70, 75, 81 and 88% after having survived 1, 2, 5 and 10 years after aHCT, respectively. The cohort was at a 13.9-fold increased risk of death compared with the general population (95% confidence interval (CI)=13.1-14.8). The risk of death approached that of the general population for 10-year survivors (standardized mortality ratio (SMR)=1.4, 95% CI=0.9-1.9), with the exception of female Hodgkin's lymphoma patients transplanted before 1995 at age 40 years (SMR=6.0, 95% CI=1.9-14.0). Among those who had survived 10 years, nonrelapse-related mortality rates exceeded relapse-related mortality rates. This study provides clinically relevant survival estimates after aHCT, and helps inform interventional strategies.

Antiviral prophylaxis has proved successful for prevention of cytomegalovirus (CMV) disease in solid organ transplant (SOT) patients; though emerging data suggest that antiviral agents interfere with immunity, and may inhibit immune priming. In this context, we investigated levels and phenotype of primary CMV-specific immune responses that developed during antiviral prophylaxis in a cohort of CMV seronegative recipients (R(-) ) of a SOT from a seropositive donor (D(+) ). We longitudinally monitored CMV viral load, antibodies and levels of the negative immuno-modulator IL-10. PBMC were stimulated with CMV-specific peptide libraries to measure CD137 activation marker on CMV-specific T-cells and levels of PD-1 receptor, which is over expressed on exhausted T-cells. Unexpectedly, the majority (13/18) of D(+) R(-) patients who developed a primary CMV response showed early post-transplant CMV-specific responses, though levels of PD-1 on CMV-specific T-cells remained elevated throughout prophylaxis. A strong inverse association was found between levels of plasma IL-10 and CMV-specific cellular immune responses. Our study suggests that during prophylaxis, subclinical CMV infection might have occurred in the D(+) R(-) patients, and primary CMV-specific responses were detected early post-transplant when levels of plasma IL-10 were low. Extended prophylaxis or antiviral treatment did not appear to suppress CMV-specific antibodies or T-cells, which, however, showed exhaustion phenotypes.

Cytomegalovirus (CMV) is the single most important viral pathogen that affects recovery of immunosuppressed transplant recipients (1). Antiviral prophylaxis for 3 months post transplant, used by ~90% of American solid organ transplant (SOT) centers, reduces the incidence of CMV disease to

Transgenic (Tg) mice expressing HLA class I alleles and lacking murine MHC class I represent a useful model for the pre-clinical evaluation of human vaccines, which focus on induction of CD8(+) T-cell responses. We have developed a platform to be used in Tg mice for exploring the immunogenicity of T-cell targets, whose immunologic epitopes have yet to be defined. To test the attributes of the evaluation system in the context of an important human pathogen, we have explored multiple antigens from cytomegalovirus (CMV). A panel of recombinant modified vaccinia Ankara (MVA) vectors, expressing various CMV proteins (CMV-MVA) was used to immunize HLA-A*0201, B*0702 and A*1101 Tg mice. Immune splenocytes were in vitro stimulated (IVS) either using syngeneic lipo-polysaccharide activated lymphoblasts or Tg HLA-I matched human EBV-transformed B-lymphoblastoid cells (LCL), both loaded with peptide libraries, encompassing the CMV protein under investigation. IVS performed with peptide library loaded lymphoblasts failed to provide a reliable stimulation. In contrast, the usage of LCL as antigen presenting cells (APC) of CMV peptide libraries resulted in a consistent and specific amplification of the Tg T-cell response in animals immunized with CMV-MVAs. The LCL IVS method reliably allowed defining the immunogenicity and immunodominant CD8(+) T-cell regions of uncharacterized CMV antigens. The combination of CMV-MVA vectors, unbiased pools of CMV-specific peptide libraries presented by Tg HLA-I matched LCL constitutes a valid tool for the pre-clinical evaluation of model candidate vaccines. This convenient method could find application to investigate the immunogenicity profile of cancer antigens or proteins from infectious human pathogens.

Cytomegalovirus (CMV)-seronegative recipients (R(-)) of a liver transplant from CMV-positive donors (D(+)) are at high risk for developing late CMV disease after discontinuation of antiviral prophylaxis. Levels of viremia and CMV-specific interferon (IFN)- gamma -producing CD4(+) and IFN- gamma -producing CD8(+) T cell responses were prospectively measured from discontinuation of antiviral prophylaxis until 1 year after transplantation in 17 consecutive D(+)/R(-) patients. CMV loads of >1000 copies/mL were strongly associated with CMV disease in the 6 symptomatic patients. Despite immunosuppression, broadly diverse T cells specific for CMV lysate or peptide libraries spanning pp65 and immediate early (IE) 1 immunodominant CMV antigens developed in all patients. A vigorous CD8(+) T cell response to pp65 and IE1 antigens characterized the D(+)/R(-) cohort. Unexpectedly, none of these responses were predictive of CMV disease or viremia. No significant lymphopenia or functional impairment of CMV-specific T cells was detected in the symptomatic patients, whose morbidity was resolved after antiviral treatment while measurable CMV immunity was maintained during the 1-year observation period.

In patients with multiple myeloma (MM) undergoing autologous hematopoietic cell transplantation (auto-HCT), peripheral blood progenitor cells (PBPCs) may be collected following mobilization with growth factor alone (GF) or cytotoxic chemotherapy plus GF ( (CC+GF). It is uncertain whether the method of mobilization affects post-transplant outcomes. We compared these mobilization strategies in a retrospective analysis of 968 patients with MM from the Center for International Blood and Marrow Transplant Research database who received an auto-HCT in the US and Canada between 2007 and 2012. The kinetics of neutrophil engraftment (≥ 0.5 × 109/L) was similar between groups (13 vs. 13 days, P=0.69) while platelet engraftment (≥ 20 × 109/L) was slightly faster with CC+GF (19 vs. 18 days, P=0.006). Adjusted 3-years PFS was 43% (95% C.I. 38–48) in GF and 40% (95% C.I. 35–45) in CC+GF, P=0.33. Adjusted 3-years OS was 82% (95% C.I. 78–86) vs. 80% (95% C.I. 75–84), P=0.43 and adjusted 5-year OS was 62% (95C.I. 54–68) vs. 60% (95% C.I. 52–67), P=0.76, for GF and CC+GF respectively. We conclude that MM patients undergoing auto-HCT have similar outcomes irrespective of the method of mobilization and found no evidence that the addition of chemotherapy to mobilization contributes to disease control.

Objective Adoptive cellular therapy of cytomegalovirus (CMV)-specific T cells in allogeneic hematopoietic stem cell transplantation (HSCT) patients is a promising approach for controlling CMV viremia and its morbidity. We sought to develop a clinically suitable strategy to dually expand infusible CD8 + and CD4 + T-cell subsets specific for CMV. Methods Polyclonal CMV T-cell lines were generated using peripheral blood mononuclear cell (PBMCs) treated with synthetic single-stranded CpG motif-containing oligodeoxynucleotides (ODNs) and infected with recombinant (r) modified vaccinia Ankara (MVA) expressing CMV antigens. Cultures derived from 12 healthy CMV-positive donors were analyzed using chromium release and lymphoproliferation assays, intracellular staining for interferon-γ (IFN-γ), and HLA tetramers. Results A 3-day incubation with a combination of ODN 2006 and 2216 was found to reproducibly generate a highly rMVA infectable population of PBMCs with concomitant high expression of CMV antigens. CpG ODN-treated autologous PBMCs infected with rMVA elicited a 30-fold average expansion of both CMV-specific CD4 + and CD8 + T cells in 10 days. The enriched T-cell populations showed minimal alloreactivity, high levels of CMV-specific HLA class I tetramer binding, cytotoxic activity, and IFN-γ production from both CD8 + and CD4 + T cells. Conclusions The ability to quickly produce autologous professional antigen-presenting cells, capable of stimulating clinically useful amounts of CMV-specific CD4 + and CD8 + T-cell lines, enhances the attractiveness of using rMVA for immunotherapeutic interventions to manage HSCT-related CMV disease.

Users’ personal information spaces are characterized by their content, organisation, and ongoing user interaction with them. They are fluid entities, evolving over time, and supporting multiple user activities that may require different perspectives of the same underlying information structure. Increasing storage capacity of computing devices and ready access to networked resources puts users at risk of information overload, and presents increasing challenges in organising and accessing their information. The hierarchical model of information organisation currently dominates personal computing, and is realised for the user in interfaces that help to manage and access filestore hierarchies. Such a model provides limited inherent support for what users do—carry out a range of interleaved activities over time. In this paper, we describe the TimeSpace system, which provides perspectives on a user’s information resources based on activities and temporal attributes of the information. TimeSpace can be used alongside, or in place of, existing systems and models (such as the Microsoft Windows hierarchical file model). User interaction with an information space is non-intrusively observed and then represented automatically in TimeSpace. Visualisations provide overviews of user activity on multiple projects and detailed views of activity within particular projects, allowing navigation forward and backward in time. An observational study of use of the system revealed positive user views of the utility of temporal, activity-oriented workspaces in real world contexts alongside existing tools. Participants appreciated being offered a different perspective on their electronic information collection, one that visually shows the composition and development of their information space. They were interested in using the system for current and long-term work as well as for archiving information, as the visualisations provide a context for their work and give an overview of all their work in progress. The ideas embodied by the system and its visualisations show promise and raise a number of issues for further exploration. In future work, these ideas will be adapted and extended to support users in managing their information spaces across multiple personal devices, locations and time.

In an attempt to decrease toxicity in high-risk patients undergoing unrelated donor hematopoietic stem cell transplantation (URD HSCT), we tested a combination of cyclosporine (CSP) and mycophenolate mofetil (MMF) as graft-versus-host disease (GVHD) prophylaxis with the reduced-intensity conditioning regimen fludarabine/melphalan (Flu/Mel). A total of 22 adult patients with advanced myeloid (n=15) and lymphoid (n=7) malignancies were treated. All patients received Flu 25 mg/m2 for 5 days and Mel 140 mg/m2, with CSP 3 mg/kg daily and MMF 15 mg/kg three times a day. The median age was 49 years (range 18-66). Durable engraftment was seen in all but one patient with myelofibrosis. The 1-year nonrelapse mortality was 32%, 27% from GVHD. The cumulative incidence of acute GVHD grade 2-4 and 3-4 was 63 and 41%, respectively. With a median follow-up of 18 months, the disease-free survival (DFS) and overall survival (OS) are 55 and 59%, respectively. For patients with AML and MDS (n=14), the DFS and OS is 71%. For patients undergoing a second transplant (n=14), the DFS and OS is 57%. In conclusion, this regimen is associated with acceptable toxicity but high rates of GVHD in high-risk patients undergoing URD HSCT. Encouraging disease control for patients with advanced myeloid malignancies was observed.

We compared the effects of calorie restriction (CR) and cyclophosphamide (CTX) on the progression of lupus nephritis and immunological changes in NZB/NZW F1 mice. Ad libitum (AL)/CTX and CR delayed onset of proteinuria and significantly decreased serum levels of anti-dsDNA, anti-histone, and circulating immune complex antibodies. CTX and CR prevented the increase in and activation of B cells, the decline in CD8+ T cells, and maintained a higher proportion of naive CD4+ and CD8+ cells. MHC class I antigen and LFA-1 expression on CD8+ T cells and MHC class II antigen on B cells were also decreased. AL/CTX and CR prevented the increase in production of IL-10 and up-regulated IL-2 production in T cells ex vivo. We concluded that both CR and CTX can delay the onset of autoimmune disease, in part by maintaining higher numbers of naive T cells and the immune responsiveness of T cells and decreasing the proportion of B cells.

The prognosis of patients with primary refractory acute myelogenous leukemia (AML) is poor. Our initial report suggested that some patients could achieve durable remission after allogeneic stem cell transplantation (SCT). Herein, we update our initial experience and report further analysis of this group of patients to determine whether there are pre-SCT prognostic factors predictive of posttransplantation relapse and survival. We reviewed the records of 68 patients who consecutively underwent transplantation at the City of Hope Cancer Center with allogeneic SCT for primary refractory AML between July 1978 and August 2000. Potential factors associated with overall survival and disease-free survival were examined. With a median follow-up of 3 years, the 3-year cumulative probabilities of disease-free survival (DFS), overall survival (OS), and relapse rate for all 68 patients were 31% (95% confidence interval [CI], 20%–42%), 30% (95% CI, 18%–41%), and 51% (95% CI, 38%–65%), respectively. In multivariate analysis, the only variables associated with shortened OS and DFS included the use of an unrelated donor as the stem cell source (relative risk, 2.23 [OS] and 2.05 [DFS]; P = .0005 and .0014, respectively) and unfavorable cytogenetics before SCT (relative risk: 1.68 [OS] and 1.58 [DFS]; P = .0107 and .0038, respectively). Allogeneic SCT can cure approximately one third of patients with primary refractory AML. Cytogenetic characteristics before SCT correlate with transplantation outcome and posttransplantation relapse.

Autologous hematopoietic stem cell transplantation (autoSCT) is an effective treatment for patients with various hematologic malignancies. Despite the significant improvement in the overall outcome, disease progression after transplantation remains the major cause of treatment failure. With longer follow-up, therapy-related myelodysplasia/acute myelogenous leukemia is becoming an important cause of treatment failure. The prognosis for these 2 groups of patients is very poor. Allogeneic hematopoietic stem cell transplantation (alloSCT) is a potential curative treatment for these patients. However, the outcome with conventional myeloablative alloSCT after failed autoSCT is typically poor because of high transplant-related mortality. In an attempt to reduce the treatment-related toxicity, we studied a reduced-intensity conditioning regimen followed by alloSCT for patients with progressive disease or therapy-related myelodysplasia/acute myelogenous leukemia after autoSCT. This report describes the outcomes of 28 patients with hematologic malignancies who received a reduced-intensity alloSCT after having treatment failure with a conventional autoSCT. Fourteen patients received a hematopoietic stem cell transplant from a related donor and 14 from an unrelated donor. The conditioning regimen consisted of low-dose (2 Gy) total body irradiation with or without fludarabine in 4 patients and the combination of melphalan (140 mg/m2) and fludarabine in 24. Cyclosporine and mycophenolate mofetil were used for posttransplantation immunosuppressive therapy, as well as graft-versus-host disease (GVHD) prophylaxis, in all patients. All patients engrafted and had >90% donor chimerism on day 100 after SCT. Currently, 13 patients (46%) are alive and disease free, 7 patients (25%) developed disease progression after alloSCT, and 8 (32%) died of nonrelapse causes. Day 100 mortality and nonrelapse mortality were 25% and 21%, respectively. With a median follow-up of 24 months for surviving patients, the 2-year probabilities of overall survival, event-free survival, and relapse rates were 56.5%, 41%, and 41.9%, respectively. Six patients (21%) developed grade III to IV acute GVHD. Among 21 evaluable patients, 15 (67%) developed chronic GVHD. We conclude that (1) reduced-intensity alloSCT is feasible and has an acceptable toxicity profile in patients who have previously received autoSCT and that (2) although follow-up was short, a durable remission may be achieved in some patients who would otherwise be expected to have a poor outcome.

Advances in effective antiretroviral therapy for HIV infection have made high-dose therapy and autologous stem cell transplantation possible in patients with HIV-associated lymphomas. Regimen-related toxicity is not significantly increased when antiretroviral therapy is combined with high-dose chemoradiotherapy. Durable engraftment can be seen with autologous stem cell rescue. Infectious complications can be managed with a combination of surveillance and prophylaxis. Long-term remissions of these high-risk lymphomas can be achieved with this approach. This suggests that patients with HIV-associated lymphomas should be considered for autologous transplantation in a manner similar to HIV-negative lymphoma patients.

Trypsin derived from either porcine or bovine pancreas is widely used in biopharmaceutical manufacturing. This industry is working toward replacing animal source materials with alternatives to prevent contamination of the products with mammalian viruses and human pathogens. Bovine trypsin made in maize seed culminated in a commercial product, TrypZean™, which is a possible replacement product for use in cell culture and viral vaccine manufacturing, as well as in the processing of therapeutic proteins. This chapter describes the development of this product from a plant biotechnology perspective and then discusses how it is being used today. Possible improvements to make this product more cost competitive are discussed.

This review addresses various aspects of HIV infection pertinent to hematology, including the consequences of HIV infection on specific aspects of hematopoiesis and an update on the current biologic, epidemiologic and therapeutic aspects of AIDS-related lymphoma and Hodgkin's disease. The results of the expanding use of progenitor cell transplantation in HIV infected patients are also reviewed. In Section I, Dr. Scadden reviews the basis for HIV dysregulation of blood cell production, focusing on the role of the stem cell in HIV disease. T cell production and thymic function are discussed, with emphasis placed upon the mechanisms of immune restoration in HIV infected individuals. Results of clinical and correlative laboratory studies are presented. In Section II, Dr. Levine reviews the recent epidemiologic trends in the incidence of lymphoma, since the widespread availability of highly active anti-retroviral therapy (HAART). The biologic aspects of AIDS-lymphoma and Hodgkin's disease are discussed in terms of pathogenesis of disease. Various treatment options for these disorders and the role of concomitant anti-retroviral and chemotherapeutic intervention are addressed. Drs. Zaia and Krishnan will review the area of stem cell transplantation in patients with AIDS related lymphoma, presenting updated information on clinical results of this procedure. Additionally, they report on the use of gene therapy, with peripheral blood CD34+ cells genetically modified using a murine retrovirus, as a means to treat underlying HIV infection. Results of gene transfer experiments and subsequent gene marking in HIV infected patients are reviewed.

The association between social classes, food intake and coronary risk factors was determined. Cross-sectional surveys were conducted in 6‐12 urban streets in each of five cities, each one from five different regions of India using similar methods of dietary intakes and criteria of diagnosis. We randomly selected 3257 women aged 25‐64 years inclusive, from Moradabad (n = 902), Trivandrum (n = 760), Calcutta (n = 410), Nagpur (n = 405) and Bombay (n = 780). All subjects, after pooling of data, were divided into social class 1 ( n = 985), class 2 (n = 790), class 3 (n = 774), class 4 (n = 602) and class 5 (n = 206) based on various attributes of socioeconomic status. Social class 1 was the highest and 5 was the lowest social class. Social classes 1‐3 had greater intake of pro-atherogenic foods; total visible fat, milk and milk products, meat and eggs, as well as sugar and confectionery, compared to social classes 4 and 5. The consumption of wheat, rice, millets, fruits, vegetables and legume/total visible fat ratio were inversely associated with social class. Mean body mass index (BMI), obesity, overweight, central obesity and sedentary lifestyle were also significantly more common among subjects from higher social classes. Spearman’s rank correlation showed that bodyweight, BMI, wheat, rice, millets, total visible fat, milk and milk products, meat, eggs, sugar and jaggery intakes were significantly correlated with social class. Social class 5 subjects had a lower intake of all foods and a lower BMI, suggestive of a higher rate of undernutrition among them. The findings indicate that the consumption of pro-atherogenic foods and other coronary risk factors are more common in higher social classes compared to lower social classes.

Objective To study the prevalence of central obesity and age-specific waist:hip ratio of urban women from five Indian cities. Design and setting Cross-sectional surveys were conducted in 6-12 urban streets in different parts of India using similar methods of sample selection and criteria of diagnosis. Subjects and methods We randomly selected 3212 women, aged 25-64 years, from Moradabad (n = 902), Trivandrum (n = 760), Calcutta (n = 365), Nagpur (n = 405), and Bombay (n = 780). Evaluation was by a questionnaire administered by a physician and a dietician, a physical examination, and anthropometric measurements. Results The overall prevalence of central obesity among the total number of women was 55.0%, with the highest prevalence in Calcutta (62.2%) and the lowest in Bombay (47.4%). Waist:hip ratio was 0.85 +/- 0.13 (mean +/- SD) with the highest ratio for women in Calcutta (0.87 +/- 0.12) and the lowest for women in Moradabad (0.84 +/- 0.16). After pooling of data from all five cities, multivariate logistic regression analysis showed that, regardless of age, body mass index (> 23 kg/m2; odds ratio 1.12), sedentary lifestyle (odds ratio 2.51), and family history of obesity (odds ratio 2.15) were strongly associated with central obesity. Excess intake of fat was weakly associated with central obesity but age was not a risk factor for central obesity, although the prevalence was highest among those aged over 55 years in Moradabad, Calcutta, and Nagpur. Conclusions The overall prevalence of central obesity among the urban women of India has increased, more so in Calcutta and Trivandrum. Body mass index, sedentary lifestyle, and family history of excess intake of fat were significant risk factors for central obesity.

Non-neovascular age-related macular degeneration (dry AMD) is a leading cause of irreversible vision loss and is associated with retinal pigment epithelium (RPE) dystrophy. The RPE monolayer is integral for maintenance of healthy photoreceptors. Many studies have shown that AMD can be recovered by cellular therapy through transplantation of RPEs into the sub-retinal space to re-establish RPE functionality and halt neurological degeneration. Here we present a cGMP compliant manufacturing process to producing transplantable RPE cells on a synthetic substrate for clinical phase I study.