Examining the effectiveness of technology use in classrooms: A tertiary meta-analysis

Identifying effective literacy instruction programs has been a focal point for governments, educators and parents over the last few decades (Ontario Ministry of Education, 2004, 2006; Council of Ontario Directors of Education, 2011). Given the increasing use of computer technologies in the classroom and in the home, a variety of information communication technology (ICT) interventions for learning have been introduced. Meta-analyses comparing the impact of these programs on learning, however, have yielded inconsistent findings (Andrews, Freeman, Hou, McGuinn, Robinson, & Zhu, 2007; Slavin, Cheung, Groff, & Lake, 2008; Slavin, Lake, Chambers, Cheung, & Davis, 2009; Torgerson & Zhu, 2003). The present tertiary meta-analytic review re-assesses outcomes presented in three previous meta-analyses. Four moderator variables assessed the impact of the systematic review from which they were retrieved, training and support, implementation fidelity and who delivered the intervention (teacher versus researcher). Significant results were found when training and support was entered as a moderator variable with the small overall effectiveness of the ICTs (ES = 0.18), similar to those found in previous research, increasing significantly (ES = 0.57). These findings indicate the importance of including implementation factors such as training and support, when considering the relative effectiveness of ICT interventions

Body condition score (BCS) and metabolic status of shelter dogs

A group of 147 shelter dogs were weighted and assigned a body condition score (BCS) using a 9 point scale system, in order to evaluate the prevalence of obesity in the kennel. More than 60% of the animals showed a BCS³6 (overweight and obese) and this condition was mainly attributed to an excess of carbohydrates and fat in the diet. In 67/147 dogs, a blood sample was drawn and the effects of BCS, age and time spent in the shelter were evaluated on biochemical parameters. Obese dogs showed significantly higher levels of triglycerides (P<0.01), while increasing BCS determined only an increasing non significant trend on cholesterol values. Age influenced creatinine (P<0.05) and the oldest dogs scoring BCS³6 registered significant higher NEFA (P<0.05) and CK (P=0.01) levels. Time spent in the shelter did not affect any parameter. The dogs' metabolic condition reflects the need of taking more care of the quality of feed administered in the shelters to avoid the negative health effects caused by chronic obesity

Clonal kinetics and single-cell transcriptional profiling of CAR-T cells in patients undergoing CD19 CAR-T immunotherapy

Chimeric antigen receptor (CAR) T-cell therapy has produced remarkable anti-tumor responses in patients with B-cell malignancies. However, clonal kinetics and transcriptional programs that regulate the fate of CAR-T cells after infusion remain poorly understood. Here we perform TCRB sequencing, integration site analysis, and single-cell RNA sequencing (scRNA-seq) to profile CD8+ CAR-T cells from infusion products (IPs) and blood of patients undergoing CD19 CAR-T immunotherapy. TCRB sequencing shows that clonal diversity of CAR-T cells is highest in the IPs and declines following infusion. We observe clones that display distinct patterns of clonal kinetics, making variable contributions to the CAR-T cell pool after infusion. Although integration site does not appear to be a key driver of clonal kinetics, scRNA-seq demonstrates that clones that expand after infusion mainly originate from infused clusters with higher expression of cytotoxicity and proliferation genes. Thus, we uncover transcriptional programs associated with CAR-T cell behavior after infusion.Published versio

Comparative use of aqueous humour 1H NMR metabolomics and potassium concentration for PMI estimation in an animal model

Estimation of the post-mortem interval (PMI) remains a matter of concern in the forensic scenario. Traditional and novel approaches are not yet able to fully address this issue, which relies on complex biological phenomena triggered by death. For this purpose, eye compartments may be chosen for experimental studies because they are more resistant to post-mortem modifications. Vitreous humour, in particular, has been extensively investigated, with potassium concentration ([K+]) being the marker that is better correlated with PMI estimation. Recently, a 1H nuclear magnetic resonance (NMR) metabolomic approach based on aqueous humour (AH) from an animal model was proposed for PMI estimation, resulting in a robust and validated regression model. Here we studied the variation in [K+] in the same experimental setup. [K+] was determined through capillary ion analysis (CIA) and a regression analysis was performed. Moreover, it was investigated whether the PMI information related to potassium could improve the metabolome predictive power in estimating the PMI. Interestingly, we found that a part of the metabolomic profile is able to explain most of the information carried by potassium, suggesting that the rise in both potassium and metabolite concentrations relies on a similar biological mechanism. In the first 24-h PMI window, the AH metabolomic profile shows greater predictive power than [K+] behaviour, suggesting its potential use as an additional tool for estimating the time since death

CD32⁺ and PD-1⁺ Lymph Node CD4 T Cells Support Persistent HIV-1 Transcription in Treated Aviremic Individuals.

A recent study conducted in blood has proposed CD32 as the marker identifying the "elusive" HIV reservoir. We have investigated the distribution of CD32 &lt;sup&gt;+&lt;/sup&gt; CD4 T cells in blood and lymph nodes (LNs) of HIV-1-uninfected subjects and viremic untreated and long-term-treated HIV-1-infected individuals and their relationship with PD-1 &lt;sup&gt;+&lt;/sup&gt; CD4 T cells. The frequency of CD32 &lt;sup&gt;+&lt;/sup&gt; CD4 T cells was increased in viremic compared to treated individuals in LNs, and a large proportion (up to 50%) of CD32 &lt;sup&gt;+&lt;/sup&gt; cells coexpressed PD-1 and were enriched within T follicular helper (Tfh) cells. We next investigated the role of LN CD32 &lt;sup&gt;+&lt;/sup&gt; CD4 T cells in the HIV reservoir. Total HIV DNA was enriched in CD32 &lt;sup&gt;+&lt;/sup&gt; and PD-1 &lt;sup&gt;+&lt;/sup&gt; CD4 T cells compared to CD32 &lt;sup&gt;-&lt;/sup&gt; and PD-1 &lt;sup&gt;-&lt;/sup&gt; cells in both viremic and treated individuals, but there was no difference between CD32 &lt;sup&gt;+&lt;/sup&gt; and PD-1 &lt;sup&gt;+&lt;/sup&gt; cells. There was no enrichment of latently infected cells with inducible HIV-1 in CD32 &lt;sup&gt;+&lt;/sup&gt; versus PD-1 &lt;sup&gt;+&lt;/sup&gt; cells in antiretroviral therapy (ART)-treated individuals. HIV-1 transcription was then analyzed in LN memory CD4 T cell populations sorted on the basis of CD32 and PD-1 expression. CD32 &lt;sup&gt;+&lt;/sup&gt; PD-1 &lt;sup&gt;+&lt;/sup&gt; CD4 T cells were significantly enriched in cell-associated HIV RNA compared to CD32 &lt;sup&gt;-&lt;/sup&gt; PD-1 &lt;sup&gt;-&lt;/sup&gt; (averages of 5.2-fold in treated individuals and 86.6-fold in viremics), CD32 &lt;sup&gt;+&lt;/sup&gt; PD-1 &lt;sup&gt;-&lt;/sup&gt; (2.2-fold in treated individuals and 4.3-fold in viremics), and CD32 &lt;sup&gt;-&lt;/sup&gt; PD-1 &lt;sup&gt;+&lt;/sup&gt; (2.2-fold in ART-treated individuals and 4.6-fold in viremics) cell populations. Similar levels of HIV-1 transcription were found in CD32 &lt;sup&gt;+&lt;/sup&gt; PD-1 &lt;sup&gt;-&lt;/sup&gt; and CD32 &lt;sup&gt;-&lt;/sup&gt; PD-1 &lt;sup&gt;+&lt;/sup&gt; CD4 T cells. Interestingly, the proportion of CD32 &lt;sup&gt;+&lt;/sup&gt; and PD-1 &lt;sup&gt;+&lt;/sup&gt; CD4 T cells negatively correlated with CD4 T cell counts and length of therapy. Therefore, the expression of CD32 identifies, independently of PD-1, a CD4 T cell population with persistent HIV-1 transcription and coexpression of CD32 and PD-1, the CD4 T cell population with the highest levels of HIV-1 transcription in both viremic and treated individuals.IMPORTANCE The existence of long-lived latently infected resting memory CD4 T cells represents a major obstacle to the eradication of HIV infection. Identifying cell markers defining latently infected cells containing replication-competent virus is important in order to determine the mechanisms of HIV persistence and to develop novel therapeutic strategies to cure HIV infection. We provide evidence that PD-1 and CD32 may have a complementary role in better defining CD4 T cell populations infected with HIV-1. Furthermore, CD4 T cells coexpressing CD32 and PD-1 identify a CD4 T cell population with high levels of persistent HIV-1 transcription

Identification and visualization of multidimensional antigen-specific T-cell populations in polychromatic cytometry data.

An important aspect of immune monitoring for vaccine development, clinical trials, and research is the detection, measurement, and comparison of antigen-specific T-cells from subject samples under different conditions. Antigen-specific T-cells compose a very small fraction of total T-cells. Developments in cytometry technology over the past five years have enabled the measurement of single-cells in a multivariate and high-throughput manner. This growth in both dimensionality and quantity of data continues to pose a challenge for effective identification and visualization of rare cell subsets, such as antigen-specific T-cells. Dimension reduction and feature extraction play pivotal role in both identifying and visualizing cell populations of interest in large, multi-dimensional cytometry datasets. However, the automated identification and visualization of rare, high-dimensional cell subsets remains challenging. Here we demonstrate how a systematic and integrated approach combining targeted feature extraction with dimension reduction can be used to identify and visualize biological differences in rare, antigen-specific cell populations. By using OpenCyto to perform semi-automated gating and features extraction of flow cytometry data, followed by dimensionality reduction with t-SNE we are able to identify polyfunctional subpopulations of antigen-specific T-cells and visualize treatment-specific differences between them

Conventional therapies deplete Brain-Infiltrating adaptive immune cells in a Mouse Model of Group 3 Medulloblastoma implicating Myeloid Cells as favorable immunotherapy targets

Medulloblastoma is the most common childhood brain cancer. Mainstay treatments of radiation and chemotherapy have not changed in decades and new treatment approaches are crucial for the improvement of clinical outcomes. To date, immunotherapies for medulloblastoma have been unsuccessful, and studies investigating the immune microenvironment of the disease and the impact of current therapies are limited. Preclinical models that recapitulate both the disease and immune environment are essential for understanding immune-tumor interactions and to aid the identification of new and effective immunotherapies. Using an immune-competent mouse model of aggressive Myc-driven medulloblastoma, we characterized the brain immune microenvironment and changes induced in response to craniospinal irradiation, or the medulloblastoma chemotherapies cyclophosphamide or gemcitabine. The role of adaptive immunity in disease progression and treatment response was delineated by comparing survival outcomes in wildtype C57Bl/6J and in mice deficient in Rag1 that lack mature T and B cells. We found medulloblastomas in wildtype and Rag1-deficient mice grew equally fast, and that craniospinal irradiation and chemotherapies extended survival equally in wildtype and Rag1-deficient mice, suggesting that tumor growth and treatment response is independent of T and B cells. Medulloblastomas were myeloid dominant, and in wildtype mice, craniospinal irradiation and cyclophosphamide depleted T and B cells in the brain. Gemcitabine treatment was found to minimally alter the immune populations in the brain, resulting only in a depletion of neutrophils. Intratumorally, we observed an abundance of Iba1+ macrophages, and we show that CD45high cells comprise the majority of immune cells within these medulloblastomas but found that existing markers are insufficient to clearly delineate resident microglia from infiltrating macrophages. Ultimately, brain resident and peripheral macrophages dominate the brain and tumor microenvironment and are not depleted by standard-of-care medulloblastoma therapies. These populations therefore present a favorable target for immunotherapy in combination with front-line treatments