Cell engineering with microfluidic squeezing preserves functionality of primary immune cells in vivo.

The translational potential of cell-based therapies is often limited by complications related to effectively engineering and manufacturing functional cells. While the use of electroporation is widespread, the impact of electroporation on cell state and function has yet to be fully characterized. Here, we use a genome-wide approach to study optimized electroporation treatment and identify striking disruptions in the expression profiles of key functional transcripts of human T cells. These genetic disruptions result in concomitant perturbation of cytokine secretion including a 648-fold increase in IL-2 secretion (P < 0.01) and a 30-fold increase in IFN-γ secretion (P < 0.05). Ultimately, the effects at the transcript and protein level resulted in functional deficiencies in vivo, with electroporated T cells failing to demonstrate sustained antigen-specific effector responses when subjected to immunological challenge. In contrast, cells subjected to a mechanical membrane disruption-based delivery mechanism, cell squeezing, had minimal aberrant transcriptional responses [0% of filtered genes misregulated, false discovery rate (FDR) q < 0.1] relative to electroporation (17% of genes misregulated, FDR q < 0.1) and showed undiminished effector responses, homing capabilities, and therapeutic potential in vivo. In a direct comparison of functionality, T cells edited for PD-1 via electroporation failed to distinguish from untreated controls in a therapeutic tumor model, while T cells edited with similar efficiency via cell squeezing demonstrated the expected tumor-killing advantage. This work demonstrates that the delivery mechanism used to insert biomolecules affects functionality and warrants further study.

Role of Radiologists in Contract Research Organizations (CROs).

Clinical trials play a vital role in advancing technology and novel therapies in the healthcare world. However, the increasing scale of trials and the complexity of the regulatory approval process is often a barrier for those interested in conducting research. Contract research organizations (CROs) aim to address this problem by offering their infrastructure and expertise to bring a therapy from conception to approval without the need for in-house staff. Clinical trial imaging often plays an essential role in this process, creating a need for radiologists and a unique opportunity to provide irreplaceable value in their ability to interpret and analyze the imaging outcomes of therapies in question. This paper explores the concept of CROs, the crucial role played by radiologists in their operation, and the nature of the CRO - radiologist relationship.

Interventions in ADHD: A comparative review of stimulant medications and behavioral therapies.

ADHD has a prevalence of approximately 10% in children with evidence supporting it's continuance into adulthood. This has a significant impact on how we address treatment at substance abuse facilities and also has implications for personal and occupational functioning. A lack of evidence to support the superiority of any one intervention over the other has created difficulties for both clinicians and parents. A recent review highlights long-term and short-term outcomes (Craig et al., 2015). This article reviews the benefits and pitfalls of both pharmacological interventions and behavioral therapies in the treatment of ADHD. Key articles were reviewed on the benefits and side effects of stimulants, the methods and benefits of behavioral interventions, and the effects of combination therapy. Google Scholar, PsychINFO, Medline, Cochrane, and CINAHL were searched with the following search words: Attention Deficit Hyperactivity Disorder, ADHD, Stimulant Medication, Behavioral Interventions, Combination Therapy, Cognitive Therapy, Functioning and Growth. It was found that stimulants are very effective during the period in which they are taken. While short term benefits are clear, longer term ones are not. Behavioral interventions play a key role for long-term improvement of executive functioning and organizational skills. There is a paucity of long-term randomized placebo controlled studies and current literature is inconclusive on what is the preferred intervention.

Are burnout prevention programs for hospital physicians needed?

INTRODUCTION: Burnout, whether as a brief episode or more protracted, affects all aspects of a physician's life. Given the critical role of physicians in society, efforts to monitor, preserve, and enhance physician health are beneficial also to their patients. We investigated the patterns of burnout in physicians. METHODOLOGY: Maslach Burnout Inventory (MBI), in the General Survey version, was handed out to interested physicians of a Canadian general hospital in a Grand Rounds lecture on "well-being of physicians." They were to complete the MBI on an anonymous basis. Fifty-five adequately completed questionnaires were received. RESULTS: Our physicians obtained significantly higher (p<0.01) average score on Emotional Exhaustion subscale of MBI (14.0, SD=7.5) and on the Cynicism subscale of the MBI than average scores from combined 9 occupational samples listed in Maslach's manual: our physicians reported more intense emotional depletion and endorsed more items suggestive of cynicism. However, their average Professional Efficacy score (27.6, SD=6.3) did not significantly differ from Maslach's combined average for this subscale. The physicians' Professional Efficacy scores were not significantly related to those of Emotional Exhaustion and of Cynicism (Pearson coefficients, p>0.05). CONCLUSIONS: This suggests that improving professional medical skills (and thus a sense of efficiency) alone cannot prevent physicians from burnout. An implementation of other preventive strategies such as those based on mindfulness or on cognitive behaviour therapy (CBT) is necessary.

Canadian Stroke Best Practice Recommendations: Telestroke Best Practice Guidelines Update 2017.

Every year, approximately 62,000 people with stroke and transient ischemic attack are treated in Canadian hospitals. The 2016 update of the Canadian Stroke Best Practice Recommendations Telestroke guideline is a comprehensive summary of current evidence-based and consensus-based recommendations appropriate for use by all healthcare providers and system planners who organize and provide care to patients following stroke across a broad range of settings. These recommendations focus on the use of telemedicine technologies to rapidly identify and treat appropriate patients with acute thrombolytic therapies in hospitals without stroke specialized expertise; select patients who require to immediate transfer to stroke centers for Endovascular Therapy; and for the patients who remain in community hospitals to facilitate their care on a stroke unit and provide remote access to stroke prevention and rehabilitation services. While these latter areas of Telestroke application are newer, they are rapidly developing, with new opportunities that are yet unrealized. Virtual rehabilitation therapies offer patients the opportunity to participate in rehabilitation therapies, supervised by physical and occupational therapists. While not without its limitations (e.g., access to telecommunications in remote areas, fragmentation of care), the evidence-to-date sets the foundation for improving access to care and management for patients during both the acute phase and now through post stroke recovery.

A phase I study of the GM-CSF antagonist E21R.

PURPOSE: E21R is a competitive inhibitor of GM-CSF. This is the initial clinical study to investigate the safety, toxicity and pharmacokinetics of escalating doses of E21R. PATIENTS AND METHODS: Cohorts of three patients received doses of 10, 30, 100, 300, 600 and 1000 micro g/kg per day given subcutaneously daily for 10 days. Eligible patients had solid tumours known to express GM-CSF receptors (breast, prostate, colon and lung cancer, and melanoma). No bone marrow involvement or concomitant steroids were permitted. A total of 22 patients received doses ranging from 10 to 1000 micro g/kg per day. There were 18 males and 4 females with a median age of 60 years (range 33 to 81 years). Eight patients had an ECOG performance status of 0, seven a performance status of 1, and seven a performance status of 2. There were ten patients with colon cancer, four with prostate cancer, three with lung cancer, three with melanoma and two with breast cancer. RESULTS: E21R was in general well tolerated and the maximum tolerated dose was not reached. The most severe toxicities were WHO grade 3 injection site erythema in one patient and grade 2 in two patients, grade 2 lethargy in three patients and grade 2 muscle aches and soreness, grade 2 joint pains and grade 2 thirst in one patient each. The primary pharmacokinetic parameters were dose-independent. Dose-dependent transient eosinophilia was noted from day 3. A fall in PSA levels was recorded in two patients with prostate cancer during their initial cycles of E21R, but they subsequently rose again. Serum from patients treated at 600 and 1000 micro g/kg per day antagonized GM-CSF-mediated TF-1 cell proliferation in vitro. CONCLUSION: E21R can be safely given at doses up to 1000 micro g/kg per day.

Electroconvulsive therapy in patients with skull defects or metallic implants: a review of the literature and case report.

CONTEXT: Head injury is often associated with psychiatric morbidity. While it is well understood that the loss of critical areas of the brain may play a role in cognitive dysfunction and change in personality, head injury can also have profound effects on mood and cognition. The role of medications in the treatment of mood disorders associated with brain injury is well documented, and there is also evidence favoring the use of electroconvulsive therapy (ECT) in this context. However, data are limited on the use of ECT in patients with skull defects or metallic head implants. EVIDENCE ACQUISITION: First, a review of the literature on use of ECT in patients with metallic head implants is provided. Electronic databases and online sites, including PubMed, Cochrane Library of Systematic Reviews, and UpToDate, were used to search for relevant articles and case reports on the use of ECT in patients with and without metallic implants in the head (1964 to 2009). The search terms electroconvulsive, electroconvulsive therapy, ECT, electroshock therapy, EST, head injury, brain injury, metallic plates, metallic implants, skull prosthesis, and depression were used interchangeably. The search produced 7 articles discussing exclusively the use of ECT in patients with a metallic skull plate. Second, the case of the successful and safe use of ECT in an individual with a previous history of brain trauma and metallic plate implantation is described. RESULTS: Most cases of head injury are managed by neurologists and rehabilitation consultants; the more severe cases of depression and other mood disorders tend to be referred for specialist psychiatric care. With greater degrees of deficit following head injury, management becomes more complicated. Our patient showed positive results with ECT, including improvement in depressive features and resolution of suicidal ideas/plans. CONCLUSION: ECT is an effective and safe alternative in patients with a history of brain trauma and metallic plate implantation who subsequently develop treatment-resistant depression and associated suicidal ideas or plans refractory to management with medications.

Osmotic stress stimulates phosphorylation and cellular expression of heat shock proteins in rhesus macaque sperm.

The cryosurvival of sperm requires cell signaling mechanisms to adapt to anisotonic conditions during the freezing and thawing process. Chaperone proteins heat shock protein 70 (HSP 70) and heat shock protein 90 (HSP 90; recently renamed HSPA and HSPC, respectively) facilitate some of these cell signaling events in somatic cells. Sperm were evaluated for their cellular expression and levels of phosphorylation of both HSP 70 and HSP 90 under anisotonic conditions as a potential model for cell signaling during the cryopreservation of macaque spermatozoa. In order to monitor the level of stress, the motility and viability parameters were evaluated at various time points. Cells were then either prepared for phosphoprotein enrichment or indirect immunocytochemistry. As controls, the phosphoserine, phosphothreonine, and phosphotyrosine levels were measured under capacitation and cryopreservation conditions and were compared with the phosphoprotein levels expressed under osmotic conditions. As expected, there was an increase in the level of tyrosine phosphorylation under capacitation and cryopreservation conditions. There was also a significant increase in the level of all phosphoproteins under hyperosmotic conditions. There was no change in the level of expression of HSP 70 or 90 under osmotic stress conditions as measured by Western blot. The enrichment of phosphoproteins followed by Western immunoblotting revealed an increase in the phosphorylation of HSP 70 but not HSP 90 under osmotic stress conditions. Indirect immunofluorescence localized HSP 70 to the postacrosomal region of sperm, and the level of membrane expression of HSP 70 was significantly affected by anisotonic conditions, as measured by flow cytometry. Taken together, these results suggest a differential role for HSP 70 and HSP 90 during osmotic stress conditions in rhesus macaque sperm.

Variability of Presentation in Hereditary Neuropathy with Liability to Pressure Palsy Results in Underrecognition.

The genetic defect responsible for hereditary neuropathy with liability to pressure palsy (HNPP) is located in the same segment that is duplicated in Charcot-Marie-Tooth type 1A (CMT1A). HNPP had been presumed to be rare until an epidemiological study found a much higher incidence than was expected; the researchers suggested that HNPP was underrecognized because many affected persons have mild symptoms. We believe that another reason for underdiagnosis of HNPP is the marked phenotypic variability of the disease. We recommend, therefore, that DNA analysis for the 17p11.2 deletion be considered in patients with unexplained demyelinating neuropathy regardless of family history.