Impairment of cognitive function in different domains early after lung transplantation

In this prospective observational pilot study patients with the diagnosis of end-stage lung disease and listed for lung transplantation underwent a cognitive function test battery before and after lung transplantation to investigate postoperative cognitive function in three domains (visual and verbal memory, executive functioning, concentration/speed of processing). Additionally we investigated intraoperative risk factors for postoperative cognitive dysfunction. In total, 24 patients were included in this pilot study. The incidence of postoperative cognitive dysfunction was 58.3%. In the cognitive dysfunction group, the domains executive functioning and concentration/attention were significantly impaired whereas memory was not affected. Patients with cognitive impairment had a significantly longer ICU stay. The strongest independent risk factor for the development of cognitive dysfunction was operation time. No influence of cerebral oxygen desaturations on cognitive dysfunction was found. This might have important implications for early psychological rehabilitation strategies in this high-risk patient collective

Mechanisms for Controlling HIV-1 Infection: A Gene Therapy Approach

Current anti-retroviral treatment (ART) for HIV-1 is highly effectively at controlling the infection. However, during early infection the virus establishes a latent reservoir, which is not impacted by ART. Any treatment interruption rapidly results in virus rebound from the latent reservoir to pre-therapy levels and thus ART does not constitute an HIV-1 cure. Alternate treatments are currently being explored in the form of gene therapy, following the success of the Berlin patient who has had undetectable virus since 2007. This review will describe the contrasting cure approaches that are currently the focus of multiple studies to control HIV, then focus in on functional cure gene therapy strategies; specifically, epigenetic silencing of HIV-1 by various methods, including RNA-directed transcriptional gene silencing. The various delivery strategies for targeting cells of the latent reservoir will be reviewed and finally, the clinical status and current challenges for ex vivo versus in vivo gene therapy delivery approaches will be discussed

Accurate isolation and detection of circulating tumor cells using enrichment-free multiparametric high resolution imaging

IntroductionThe reliable and accurate detection of rare circulating tumor cells (CTCs) from cancer patient blood samples promises advantages in both research and clinical applications. Numerous CTC detection methods have been explored that rely on either the physical properties of CTCs such as density, size, charge, and/or their antigen expression profiles. Multiple factors can influence CTC recovery including blood processing method and time to processing. This study aimed to examine the accuracy and sensitivity of an enrichment-free method of isolating leukocytes (AccuCyte® system) followed by immunofluorescence staining and high-resolution imaging (CyteFinder® instrument) to detect CTCs.MethodHealthy human blood samples, spiked with cancer cells from cancer cell lines, as well as blood samples obtained from 4 subjects diagnosed with cancer (2 pancreatic, 1 thyroid, and 1 small cell lung) were processed using the AccuCyte-CyteFinder system to assess recovery rate, accuracy, and reliability over a range of processing times.ResultsThe AccuCyte-CyteFinder system was highly accurate (95.0%) at identifying cancer cells in spiked-in samples (in 7.5 mL of blood), even at low spiked-in numbers of 5 cells with high sensitivity (90%). The AccuCyte-CyteFinder recovery rate (90.9%) was significantly higher compared to recovery rates obtained by density gradient centrifugation (20.0%) and red blood cell lysis (52.0%). Reliable and comparable recovery was observed in spiked-in samples and in clinical blood samples processed up to 72 hours post-collection. Reviewer analysis of images from spiked-in and clinical samples resulted in high concordance (R-squared value of 0.998 and 0.984 respectively).DiscussionThe AccuCyte-CyteFinder system is as an accurate, sensitive, and clinically practical method to detect and enumerate cancer cells. This system addresses some of the practical logistical challenges in incorporating CTCs as part of routine clinical care. This could facilitate the clinical use of CTCs in guiding precision, personalized medicine

Magnetic resonance imaging (MRI) contrast agents for tumor diagnosis

10.1260/2040-2295.4.1.23Journal of Healthcare Engineering4123-4

Delivery of RNA therapeutics for an HIV-1 gene therapy cure

Due to the success of antiretroviral therapy (ART), people living with HIV now experience a controlled chronic disease, compared to uncontrolled disease off-ART that progresses to AIDS. Although ART has revolutionized HIV-1 treatment, it is not a cure. This is due to the latent reservoir that can rebound to pre-therapy levels upon ART cessation. Successful HIV-1 stem cell therapy cures have set a precedent for targeting the HIV-1 co-receptor CCR5 (i.e. the Berlin and London patients). A complementary HIV-1 functional cure strategy is the ‘block and lock’ strategy, where, for example, small interfering (si)RNA targeting the promoter block virus transcription and lock the promoter in a latent state via epigenetic silencing. This thesis will assess a combined gene therapy cure incorporating CCR5 targeting and RNAi-induced epigenetic silencing. A major barrier for RNAi-directed HIV-1 gene therapy is efficient delivery of siRNA into cells. This study assessed two different delivery platforms; nanoparticles and lentiviral vectors. Numerous nanoparticle formulations were screened and identified a lead candidate that successfully delivered siRNA in cell lines and human primary cells in vitro and induced functional silencing of virus replication. In parallel, delivery of shRNAs with an established lentiviral delivery system was assessed. Studies confirmed a novel epigenetic silencing shRNA, sh136/T, and demonstrated successful combination delivery of two shRNAs in a single lentiviral construct (i.e. shPromA and shCCR5). Reduced virus reactivation was reported in multiple J-Lat latent cell lines and shRNAs suppressed virus replication in PM1 cells. In vivo studies were performed in a chronic HIV-1 humanized mouse model following engraftment of shPromA or shPromA-shCCR5 transduced CD34+ hematopoietic stem cells (HSCs). Following HIV-1 infection for 10 weeks and ART treatment for 8, virus rebound was followed for 28 days post-ART treatment interruption. Mice carrying the shPromA-shCCR5 gene modification demonstrated increased CD4+ T cell count and delayed viral rebound post-ART treatment interruption. This thesis provides valuable insights for si/shRNA therapeutic delivery using nanoparticle delivery in primary human cells in vitro and lentiviral delivery in T cell lines (in vitro) and primary CD34+ HSCs (in vivo). Both delivery platforms were capable of delivering si/shRNAs to functionally suppress virus replication

Early childhood professionals' perceptions of children's school readiness characteristics in six countries

The opinions of early years educators and primary school teachers regarding the school readiness characteristics (SRC) that best support a child's positive transition to school differ. The aims of our study were to determine key elements of perceived SRC and to identify the priorities of pre-service and in-service early years professionals in six countries: Australia, Austria, Colombia, Germany, Nicaragua, and Slovenia. In total, 1198 early years professionals responded to a survey that investigated perceptions of child school readiness characteristics. Independence, social skills and concentration were reported to be very important. Academic precursors and physical development were reported to be the least important SRC. Findings for the total sample and specific subgroups are presented and discussed

NRF2/Itaconate Axis Regulates Metabolism and Inflammatory Properties of T Cells in Children with JIA

Background: CD4+ T cells critically contribute to the initiation and perturbation of inflammation. When CD4+ T cells enter inflamed tissues, they adapt to hypoxia and oxidative stress conditions, and to a reduction in nutrients. We aimed to investigate how this distinct environment regulates T cell responses within the inflamed joints of patients with childhood rheumatism (JIA) by analyzing the behavior of NRF2—the key regulator of the anti-oxidative stress response—and its signaling pathways. Methods: Flow cytometry and quantitative RT-PCR were used to perform metabolic profiling of T cells and to measure the production of inflammatory cytokines. Loss of function analyses were carried out by means of siRNA transfection experiments. NRF2 activation was induced by treatment with 4-octyl-Itaconate (4-OI). Results: Flow cytometry analyses revealed a high metabolic status in CD4+ T cells taken from synovial fluid (SF) with greater mitochondrial mass, and increased glucose and fatty acid uptake. This resulted in a heightened oxidative status of SF CD4+ T cells. Despite raised ROS levels, expression of NRF2 and its target gene NQO1 were lower in CD4+ T cells from SF than in those from blood. Indeed, NRF2 activation of CD4+ T cells downregulated oxidative stress markers, altered the metabolic phenotype and reduced secretion of IFN-γ. Conclusion: NRF2 could be a potential regulator in CD4+ T cells during chronic inflammation and could instigate a drift toward disease progression or regression, depending on the inflammatory environment