The CellPhe toolkit for cell phenotyping using time-lapse imaging and pattern recognition.

With phenotypic heterogeneity in whole cell populations widely recognised, the demand for quantitative and temporal analysis approaches to characterise single cell morphology and dynamics has increased. We present CellPhe, a pattern recognition toolkit for the unbiased characterisation of cellular phenotypes within time-lapse videos. CellPhe imports tracking information from multiple segmentation and tracking algorithms to provide automated cell phenotyping from different imaging modalities, including fluorescence. To maximise data quality for downstream analysis, our toolkit includes automated recognition and removal of erroneous cell boundaries induced by inaccurate tracking and segmentation. We provide an extensive list of features extracted from individual cell time series, with custom feature selection to identify variables that provide greatest discrimination for the analysis in question. Using ensemble classification for accurate prediction of cellular phenotype and clustering algorithms for the characterisation of heterogeneous subsets, we validate and prove adaptability using different cell types and experimental conditions.

Sodium accumulation in breast cancer predicts malignancy and treatment response.

BACKGROUND: Breast cancer remains a leading cause of death in women and novel imaging biomarkers are urgently required. Here, we demonstrate the diagnostic and treatment-monitoring potential of non-invasive sodium (23Na) MRI in preclinical models of breast cancer. METHODS: Female Rag2-/- Il2rg-/- and Balb/c mice bearing orthotopic breast tumours (MDA-MB-231, EMT6 and 4T1) underwent MRI as part of a randomised, controlled, interventional study. Tumour biology was probed using ex vivo fluorescence microscopy and electrophysiology. RESULTS: 23Na MRI revealed elevated sodium concentration ([Na+]) in tumours vs non-tumour regions. Complementary proton-based diffusion-weighted imaging (DWI) linked elevated tumour [Na+] to increased cellularity. Combining 23Na MRI and DWI measurements enabled superior classification accuracy of tumour vs non-tumour regions compared with either parameter alone. Ex vivo assessment of isolated tumour slices confirmed elevated intracellular [Na+] ([Na+]i); extracellular [Na+] ([Na+]e) remained unchanged. Treatment with specific inward Na+ conductance inhibitors (cariporide, eslicarbazepine acetate) did not affect tumour [Na+]. Nonetheless, effective treatment with docetaxel reduced tumour [Na+], whereas DWI measures were unchanged. CONCLUSIONS: Orthotopic breast cancer models exhibit elevated tumour [Na+] that is driven by aberrantly elevated [Na+]i. Moreover, 23Na MRI enhances the diagnostic capability of DWI and represents a novel, non-invasive biomarker of treatment response with superior sensitivity compared to DWI alone.

Volatile compounds in human breath: critical review and meta-analysis.

Volatile compounds contained in human breath reflect the inner workings of the body. A large number of studies have been published that link individual components of breath to disease, but diagnostic applications remain limited, in part due to inconsistent and conflicting identification of breath biomarkers. New approaches are therefore required to identify effective biomarker targets. Here, volatile organic compounds have been identified in the literature from four metabolically and physiologically distinct diseases and grouped into chemical functional groups (e.g. methylated hydrocarbons or aldehydes; based on known metabolic and enzymatic pathways) to support biomarker discovery and provide new insight on existing data. Using this functional grouping approach, principal component analysis doubled explanatory capacity from 19.1% to 38% relative to single individual compound approaches. Random forest and linear discriminant analysis reveal 93% classification accuracy for cancer. This review and meta-analysis provides insight for future research design by identifying volatile functional groups associated with disease. By incorporating our understanding of the complexities of the human body, along with accounting for variability in methodological and analytical approaches, this work demonstrates that a suite of targeted, functional volatile biomarkers, rather than individual biomarker compounds, will improve accuracy and success in diagnostic research and application.

Using Simulation to Train Medical Units for Deployment.

INTRODUCTION: Recent downsizing and budgeting issues have led to challenges when attempting to train military health care reserve forces. A specific military unit collaborated with a university simulation center in order to provide more meaningful training experiences that mirrored the deployment operational experience. METHODS: The article discusses the processes used to initiate and build a simulation-based military health care training curriculum. The team consisted of an educator specializing in the use of simulation and military content experts. Rubrics for all skills were developed or found in nursing or emergency medical technician/Paramedic textbooks. Skills station training was completed using deliberate practice, where students practiced until they reached mastery level. Simulation scenarios were completed that included a primary and secondary survey improvised explosive device event. Simulation curriculum has expanded to include psychiatric scenarios using standardized patients (actors). RESULTS: Documentation of the training yielded greater insight into the unit's abilities, strengths and weaknesses. After the initial 1-year time period, 87% of medical technicians and 92% of registered nurses completed training. In total, 12 out of 38 participants needed additional support. The participants reported through the Debriefing Assessment for Simulation in Healthcare tool that they enjoyed and valued the meaningful training. CONCLUSION: Training using simulation has been invaluable to improving team cohesiveness. Building a training curriculum has generated a new perspective on ways in which military units may train and assess the strengths and opportunities of the unit. The ability to see participants in action allowed for a clearer understanding of the knowledge and skill each participant actually possessed versus what was assumed. The information obtained was invaluable to leadership in determining the true readiness of the unit for deployment. The authors offer the scenarios, lesson plans, and curriculum that they developed to other units that are interested in the training.

Pharmacokinetic and clinical outcomes when ideal body weight is used to dose busulfan in obese hematopoietic stem cell transplant recipients.

Weight-based dosing of intravenous busulfan is widely used in hematopoietic cell transplantation. However, a variety of dosing weights have been described. The objective of this retrospective study was to determine the pharmacokinetic impact of using ideal body weight as the initial dosing weight in obese as compared to non-obese transplant recipients. The secondary objectives were to describe the use of alternative dosing weights, the impact on survival, and the rates of toxicities. The mean steady-state concentration was 779.3 ng/mL (n = 82) in the non-obese cohort and 673.7 ng/mL (n = 63) in the obese cohort (p < 0.001). A smaller proportion of concentrations were below goal in the non-obese cohort (10% vs. 41%, p < 0.001). Ideal body weight and adjusted body weights with a 25 and 40% correction factor are appropriate in non-obese patients; adjusted body weights with a 25 and 40% correction factor are appropriate in obese patients. There was no difference in overall survival (p = 0.18); there was a difference in median progression-free survival (1078 vs. 500 days, p = 0.045) in the non-obese compared to obese cohorts. The use of ideal body weight to dose busulfan resulted in lower steady-state concentrations, a larger proportion of subtherapeutic concentrations, and worse progression-free survival in obese patients.

Using Evidence-Based Best Practices of Simulation, Checklists, Deliberate Practice, and Debriefing to Develop and Improve a Regional Anesthesia Training Course.

The 2011 Institute of Medicine report on the future of nursing recommended that nurses practice to the full extent of their education and training. Nurse anesthetists in certain regions of the country have been unable to maintain regional anesthesia skills because of anesthesia practice models. Factors including increased patient loads, economic motivators, and desire to maintain skill sets are driving evolution of the anesthesia practice model. In many practices, Certified Registered Nurse Anesthetists (CRNAs) now have the opportunity to expand their practice scope to include regional anesthesia. This has created the need for a pathway to rapidly develop or augment skills for CRNAs who have not been performing regional anesthesia. Well-designed and facilitated simulation methods can be effective for teaching and evaluating clinical skills with incorporation of rigorous assessment instruments to ensure consistency in training outcomes. The purpose of this quality improvement project was to determine the effectiveness of a blended-learning regional anesthesia training curriculum on improving CRNA knowledge, skill, and attitude in regional anesthesia administration as part of a clinical credentialing pathway. Forty-nine CRNAs completed all course components, including meeting all skill training thresholds through deliberate practice and use of validated checklists. Knowledge and confidence levels demonstrated significant gains.

Tandem Autologous Stem Cell Transplantation for Multiple Myeloma Patients Based on Response to Their First Transplant-A Prospective Phase II Study.

In this prospective phase II clinical trial, multiple myeloma (MM) patients were randomized to receive a second (tandem) autologous stem cell transplantation (ASCT) based on whether they achieved a partial response or worse (≤PR) following initial ASCT (ASCT1). Patients who achieved a very good partial response or better (≥VGPR) had salvage ASCT at relapse. Seventy-five patients received conditioning therapy and ASCT1. A total of 44 patients (59%) achieved ≥VGPR, whereas 31 patients entered ≤PR and were offered tandem ASCT. In all, 20 patients agreed to tandem ASCT. Demographic and clinical characteristics were similar between the two cohorts except for median lactate dehydrogenase (LDH) (P = 0.0141) and percentage of marrow plasma cells before ASCT1 (P = 0.0047), both lower in the ≥VGPR group. Intent to treat analysis showed that patients who achieved ≥VGPR to ASCT1 had a trend toward improved progression-free survival (PFS) (37 vs. 26 months, P = 0.078) and superior overall survival (OS) (not reached vs. 50 months, P = 0.0073). Patients with ≤PR who declined tandem transplantation had shortened PFS (20 vs. 28 months, P = 0.05) but similar OS (53 vs. 57.5 months, P = 0.29) compared to those who received it. Thus, a favorable clinical response to ASCT1 identifies a low-risk group with superior long-term prognosis despite similar PFS.

Hematopoietic cell transplantation in 2020: summary of year 2 recommendations of the National Marrow Donor Program's System Capacity Initiative.

The National Marrow Donor Program, in partnership with the American Society for Blood and Marrow Transplantation, sponsored and organized a series of symposia to identify complex issues affecting the delivery of hematopoietic cell transplantation (HCT) and to collaboratively develop options for solutions. "Hematopoietic Cell Transplantation in 2020: A System Capacity Initiative" used a deliberative process model to engage professional organizations, experts, transplant centers, and stakeholders in a national collaborative effort. Year 2 efforts emphasized data analysis and identification of innovative ideas to increase HCT system efficiency, address future capacity requirements, and ensure adequate reimbursement for HCT programs to meet the projected need for HCT. This report highlights the deliberations and recommendations of Year 2 and the associated symposium held in September 2011.

A combination of Botulinum Toxin A therapy and Functional Electrical Stimulation in children with cerebral palsy--a pilot study.

BACKGROUND: Among the ambulant population of children with spastic cerebral palsy (CP), dynamic equinus is one of the most common form of gait deviation that is encountered. OBJECTIVE: To investigate the combined effects of Functional Electrical Stimulation (FES) and Botulinum Toxin A (BTXA) therapy in children with spastic CP, and to demonstrate the feasibility of this combination therapy. METHODS: A single-subject design with repeated measures was adopted. Eight children (six males, two females; mean age 7 y 9 mo, SD 1 y 5 mo; range 7 y to 11 y) diagnosed with hemiplegic (n=6) or diplegic (n=2) spastic CP completed the study. Each subject participated in the study for twenty weeks. This period consisted of baseline (one week), BTXA phase (three weeks), first FES phase (four weeks), first control phase (four weeks), second FES phase (four weeks) and second control phase (four weeks). Subjects were assessed at the end of each phase. The ankle angle at the end of swing phase was selected as the primary outcome measure. The secondary outcome measure recorded was the foot contact pattern. RESULTS: There was an increase in ankle dorsiflexion at the end of the combined intervention in most subjects (n=6), accompanied by an improvement in foot contact pattern. CONCLUSIONS: This pilot study demonstrated that it is feasible to combine BTXA therapy with FES in ambulant children with spastic CP.

African American adult apheresis donors respond to granulocyte-colony-stimulating factor with neutrophil and progenitor cell yields comparable to those of Caucasian and Hispanic donors.

BACKGROUND: Granulocyte-colony-stimulating factor (G-CSF)-mobilized peripheral blood progenitor cells (PBPCs) are the most common source of cells used for hematopoietic transplantation. Benign ethnic neutropenia has been found in persons of African descent, affecting circulating white blood cells (WBCs), but not WBC production within marrow. Persons of African descent have reduced neutrophil mobilization after steroid administration, and newborns have fewer nucleated and progenitor cells in their cord blood. STUDY DESIGN AND METHODS: Twenty-two African American (AA) and 12 Hispanic PBPC donors were age, sex, and weight matched with 34 Caucasian donors. Groups were compared based on WBC and neutrophil counts after mobilization and numbers of CD34+ cells collected on Day 5 of G-CSF mobilization. RESULTS: AA donors had significantly lower baseline WBC (6.1±1.1 vs. 7.1±1.7, p=0.04) and neutrophil (3.4±1.1 vs. 4.5±1.3, p=0.01) counts compared to matched Caucasian donors. G-CSF-stimulated AAs had a significantly greater increase in WBC and neutrophil counts compared to matched Caucasians (889±293% vs. 665±230% neutrophils, p=0.02). There was no significant difference in product cell counts when comparing total nucleated, CD3+, CD34+, and mononuclear cells or colony-forming units (CFUs) between Caucasians and Hispanics or AAs and trends to greater numbers of neutrophils in products from AA donors. CONCLUSION: When stimulated by G-CSF, AAs are able to increase WBC and neutrophil counts to a higher degree than Caucasians, achieving similar numbers of neutrophil and progenitor cells in apheresis products despite starting from lower baseline blood counts.

Hypomethylating agent induction therapy followed by hematopoietic cell transplantation is feasible in patients with myelodysplastic syndromes.

Disease remission in patients with myelodysplastic syndromes can be achieved with azanucleosides, which act as pyrimidine analogs and hypomethylating agents. However, despite treatment with azanucleoside induction, patients with myelodysplastic syndromes nearly always relapse. Allogeneic hematopoietic cell transplantation (HCT) can be curative, but it is risky. Given that azanucleosides affect human leukocyte antigen expression and lymphocyte reactivity, we conducted a retrospective study to define the impact of pre-HCT azanucleoside therapy on post-HCT donor chimerism. Patients receiving azanucleoside induction therapy achieved rapid and high levels of donor chimerism post-transplant. Lineage analysis also found rapid donor chimerism of lymphocyte and granulocyte subsets. These data indicate the feasibility of pretransplant azanucleoside therapy in patients who subsequently receive an HCT.

Managing manpower and cutting costs in the health care industry.

The Bureau of Labor Statistics projects that health care services will account for one out of every six new jobs from 2002 to 2012. Based upon workload fluctuations, some companies in health care have opted to utilize "just-in-time" employees. Such an employee not only serves to stabilize the workforce but can also reduce employers' cost by allowing them to pay for labor only when they need it. Based on the analysis, a company should reduce reliance on casual staff, as the upfront cost per hire is far greater than hiring a temporary employee. Information presented points to fairly high turnover among casual employees, thus bolstering the argument against this staffing scheme when compared with temporary employee staffing.

Phase 1 study of TLK286 (Telcyta) administered weekly in advanced malignancies.

PURPOSE: To determine the dose-limiting toxicities, maximum tolerated dose, and pharmacokinetics of TLK286, a novel cancer prodrug, administered weekly. PATIENTS AND METHODS: Patients with advanced malignancies were treated with TLK286 administered weekly by i.v. infusion over 30 min in escalating doses 60-960 mg/m(2). A treatment cycle was defined as 3 weekly treatments. Patients underwent tumor assessments on day 43, and those patients receiving clinical benefit continued on treatment until disease progression or unacceptable toxicity. Safety was assessed by the WHO criteria. RESULTS: Thirty-seven patients received 111 cycles of TLK286 at eight dose levels (median, 3 cycles; range, 1-16 cycles). In this study, TLK286 given weekly at 960 mg/m(2) was well tolerated without dose-limiting toxicities. TLK286-related toxicities included grade 1-2 nausea and vomiting, fatigue and anemia. Nine of 31 evaluable patients continued therapy beyond day 43 and received a median of 5 cycles (range of 3-16 cycles) and experienced durable stable disease or minor tumor regression. Pharmacokinetic characteristics of TLK286 are described by an optimized two-compartment model. Mild to moderate renal or hepatic organ dysfunction did not impact the elimination of TLK286. CONCLUSIONS: TLK286 administered weekly at doses up to 960 mg/m(2) were well tolerated. The safety and antitumor activity observed in a broad range of cancer types supports Phase 2 disease-specific investigations of TLK286 given weekly at 960 mg/m(2).