News Article | February 15, 2017
Thailand has become the first Asian country to eliminate mother-to-child transmission (MTCT) of HIV, thanks to a pragmatic multi-sector response backed by strong political commitment and heavy government investment, a study published in Paediatrics and International Child Health reports. Such an early, concerted response allowed the country to successfully address the four prongs of the recommended World Health Organization (WHO) elimination strategy. As a result, MTCT rates were reduced from 20-40% in the mid-1990s to 1.9% in 2015 (surpassing the WHO elimination target of The WHO strategy focuses on the following four prongs: primary prevention of HIV in women of childbearing age; prevention of unintended pregnancies in women living with HIV; prevention of HIV transmission from an HIV-infected woman to her infant; and provision of appropriate treatment, care and support to women and children living with HIV. In Thailand, initiatives to promote condom use, provide information about the risk of transmission and introduce testing for pregnant and post-partum women were successfully implemented. For example, the 100% Condom Programme, which promotes 100% condom use by male patrons of commercial sex workers, has played a crucial role in preventing HIV infection in women of reproductive age. The success of such initiatives resulted in part from strong political leadership - the national AIDS policy of Thailand was transferred from the Ministry of Public Health to the Office of the Prime Minister in 1991 - and greatly increased investment, with government spending on the HIV/AIDS programme rising from US$684,000 in 1988 to US$82 million by 1997. The high rate of antenatal care provision in Thailand is also key. A voluntary HIV test with same-day results is offered at the first clinic visit, followed by re-testing later in pregnancy for HIV-negative women. For HIV-infected pregnant women, antiretroviral therapy (ART) is provided as soon as possible. Such treatment is now available at much lower cost, thanks to legislative changes which have allowed the non-commercial production of generic ART in Thailand. Counselling services at antenatal clinics also promote the use of dual methods of contraception to prevent unintended pregnancy in women with HIV. The study's author, Professor Usa Thisyakorn of Chulalongkorn University, Bangkok said: "Thailand has achieved WHO elimination of mother-to-child HIV transmission targets with early and concerted efforts of all sectors of Thai society. This provided numerous lessons learned in working together to safeguard children. Since children are the country's future, how the country responds to the problems created for them indicates how highly the country values its future." When referencing the article: Please include Journal title, author, published by Taylor & Francis and the following statement: Please note the article will not be available online until the embargo has been lifted. Taylor & Francis Group partners with researchers, scholarly societies, universities and libraries worldwide to bring knowledge to life. As one of the world's leading publishers of scholarly journals, books, ebooks and reference works our content spans all areas of Humanities, Social Sciences, Behavioural Sciences, Science, and Technology and Medicine. From our network of offices in Oxford, New York, Philadelphia, Boca Raton, Boston, Melbourne, Singapore, Beijing, Tokyo, Stockholm, New Delhi and Johannesburg, Taylor & Francis staff provide local expertise and support to our editors, societies and authors and tailored, efficient customer service to our library colleagues. For more information please contact: Sayjal Mistry, Press & Media Relations Coordinator email: email@example.com Follow us on Twitter: @tandfnewsroom Introducing Press Pass: journalist access to all Taylor & Francis Journal articles. Contact us for more details.
News Article | August 22, 2016
The study protocols were approved by University of California San Diego and Salk Institute IRB/ESCRO committees (protocols 141223ZF and 95-0001, respectively). Four TD individuals (ages 8–19 years) and five individuals with WS (ages 8–14 years; Extended Data Fig. 1a) were included in the analysis: four of the latter group had typical WS gene deletions and one (pWS88) had a partial deletion in the WS region. Informed consents were obtained from all participants or their parents as appropriate. Genetic diagnosis of WS was established using fluorescent in situ hybridization probes for elastin (ELN), a gene consistently associated with the deletion in the typical WS region1, 9. All of the participants with WS having confirmed genetic deletion exhibited the medical and clinical characteristics of the WS phenotype, including previously established cognitive, behavioural and physical features associated with the syndrome4. A diagnosis of WS was confirmed on the basis of the Diagnostic Score Sheet (DSS) for WS (American Academy of Paediatrics Committee on Genetics, 2001), with a particular focus on the cardiovascular abnormalities and the characteristic facial features associated with the ELN deletion. The scores for the participants were at the mean for WS (9) or higher, with the individual with partial deletion in the WS chromosomal region (pWS88) scoring lower than the individuals with typical WS deletion. Similarly, pWS88 reported fewer symptoms with connective tissue and growth, his cognitive scores were slightly higher than the typical individuals with WS, and he did not demonstrate the disparity between verbal and visual–spatial abilities typical of WS. However, pWS88 did display behavioural and developmental features consistent with WS, including developmental delay, over-friendliness and anxiousness. The participants were administered standard tests to quantify their non-verbal and verbal abilities, as well as versions of the WS cognitive and social profiles to capture the distinct pattern of strengths and weaknesses both within and across domains associated with the WS cognitive and social phenotype. Details of the tests and the measures tapping into the two profiles are presented in Extended Data Fig. 1. The WS cognitive profiles for the five participants with WS were constructed by calculating the log of predictive likelihood ratios under assumed normality for age-appropriate TD versus WS classifications on the basis of verbal and performance IQ (VIQ and PIQ), Beery Developmental Test of Visual-Motor Integration (VMI) and Peabody Picture Vocabulary Test (PPVT) standard scores, subject to availability. Predictive distributions were based on the published normative mean and s.d. for each of the tests employed, whereas for the WS classification the predictive distributions26 were determined using data from n = 81 (VIQ and PIQ), n = 56 (VMI) and n = 97 (PPVT) participants in a broader WS sample (described in Extended Data Fig. 1d). A tobit model was used to estimate parameters for individuals with WS on the VMI owing to the presence of floor effects. The WS social profiles for the five participants with WS were constructed using measures of social approach behaviour, emotionality/empathy and language use. Quantitative PCR was used to define the breakpoints of deleted regions in DNA isolated from iPSCs, or lymphoblast cell lines for participants with WS, with probes spanning from CALN1 to WBSCR16 and template DNA. Taqman expression assay probes detecting the WS region genes were designed and synthesized with sequences shown in Supplementary Table 11. RNase P (VIC) was used as control. Quantitative PCR was performed on the ABI PRISM 7900HT system and the results were analysed using SDS 3.2. We avoided invasive sample collection methods such as skin biopsy or blood withdrawal by taking advantage of the natural loss of deciduous teeth as a source of somatic cells. We chose to reprogram dental pulp cells (DPCs) because these cells develop from the same set of early progenitors that generate neurons. Furthermore, the neurons derived from iPSCs generated from DPCs express higher levels of forebrain genes compared with those generated from skin fibroblast-derived iPSCs27, serving the purpose of this study. Deciduous teeth were collected when they fell out and were shipped to our laboratory in DMEM 1× (Mediatech) with 4% Pen/Strep (Mediatech). Dental pulp was pulled out, washed in PBS with 4% Pen/Strep and incubated in 5% TrypLE (Gibco) for 15 min. Pulp was partly dissociated using needles and plated in culture medium (DMEM/F12 50:50, 15% FBS, 1%NEAA, 1% fungizone and 2% Pen/Strep). In 1–4 weeks, DPCs migrated out of the pulp and could be passaged and frozen as stock. DPCs in early passage (two to three) were reprogrammed using pMXs retroviruses expressing Yamanaka transcription factors (obtained from Addgene, Cambridge, Massachusetts)12. After 4 days, transduced DPCs were trypsinized, plated on mouse embryonic fibroblasts and cultured using human embryonic stem cell (hESC) medium. After manually picked and clonally expanded, feeder-free iPSCs were grown on matrigel-coated dishes (BD Bioscience, San Jose, California) with mTeSR1 (StemCell Technologies) or iDEAL28. All G-banding karyotyping analyses were performed by Molecular Diagnostics Service (San Diego, California) and Children’s Hospital Los Angeles (Los Angeles, California). Two hundred nanograms of DNA were processed and hybridized to the Illumina Infinium Human Core Exome BeadChip following manufacturer’s instructions. Illumina GenomeStudio V2011.1 with the Genotyping Module version 1.9.4 was used to normalize data and call genotypes using reference data provided by Illumina. Illumina’s cnv Partition and gada R packages were used to automatically detect aberrant copy number region. In addition, the B Allele Frequency (BAF) and Log R Ratio (LRR) distributions were manually checked to determine additional CNVs not detected by the software. Sample identification/relatedness was assessed by comparing called genotypes for each sample. The absolute number of different genotypes was counted and the Euclidean distances were calculated to identify relatedness of the samples. Dissociated iPSC colonies were centrifuged and resuspended in 1:1 matrigel and phosphate buffer saline solution. The cells were injected subcutaneously in nude mice. After 1–2 months, teratomas were dissected, fixed and sliced. Sections were stained with haematoxylin and eosin for further analysis. Protocols were previously approved by the University of California San Diego Institutional Animal Care and Use Committee. iPSCs were cultured on matrigel-coated dishes and fed daily with mTeSR for 7 days. On the next day, mTeSR was substituted by N2 medium (DMEM/F12 supplemented with 0.5× N2-Supplement (Life Technologies), 1 μM dorsomorphin (Tocris) and 1 μM SB431542 (Stemgent)) for 1–2 days. iPSC colonies were lifted off, cultured in suspension on the shaker (95 r.p.m. at 37 °C) for 8 days to form embryoid bodies and fed with N2 media. Embryoid bodies then were mechanically dissociated, plated on a matrigel-coated dish and fed with N2B27 medium (DMEM/F12 supplemented with 0.5× N2-Supplement, 0.5× B27-Supplement (Life Technologies), 1% penicillin/streptomycin and 20 ng/mL FGF-2). The emerging rosettes were picked manually, dissociated completely using accutase and plated on a poly-ornithine/laminin-coated plate. NPCs were expanded in N2B27 medium and fed every other day. To differentiate NPCs into neurons, FGF-2 was withdrawn from the N2B27 medium. NPCs and neurons were characterized for stage-specific markers by immunostaining and flow cytometry (NPCs only), expression profile by single-cell RT–PCR and RNA sequencing and electrophysiological property (neurons). Total RNA of DPCs, iPSCs, NPCs and neurons was extracted using TRIzol reagent (Life Technologies) according to the manufacturer’s protocols. Contaminating DNA in RNA samples was removed using TURBO DNase (Life Technologies) according to the manufacturer’s protocols. Quality and quantity of DNase-treated RNA were assessed using NanoDrop 1000 (Thermo Scientific). RNA was extracted from iPSCs as previously described using Trizol reagent (Life Technologies). cDNA was generated from the RNA using SuperScript III protocol according to the manufacturer’s instructions. PCR was performed using primers listed below at the following cycles: 94 °C for 10 min; 35 repeats of 94 °C for 30 s, 62 °C for 30 s and 72 °C for 1 min; and finally, 72 °C for 7 min. As a positive control, the pMX plasmid of the four vectors used on the reprogramming of the cells was placed along the samples as well as water as a negative template control for amplification. As an additional positive control for the endogenous genes, two hESC lines were used along with our iPSCs: H1 and HUES6 cells. Primers used were as follows. Endo-cMyc: forward, TTG AGG GGC ATC GTC GCG GGA; reverse, GCG TCC TGG GAA GGG AGA TCC. Endo-Klf4: forward, GAA ATT CGC CCG CTC CGA TGA; reverse, CTG TGT GTT TGC GGT AGT GCC. Endo-OCT3/4: forward, TCT TTC CAC CAG GCC CCC GGC TC; reverse, TGC GGG CGG ACA TGG GGA GAT CC. Endo-SOX2: forward, GCC GAG TGG AAA CTT TTG TCG; reverse, GGC AGC GTG TAC TTA TCC TTC T. Exo transgenes pMXs-TgUS: forward, GTG GTG GTA CGG GAA ATC AC. Exo-Oct4 pMXs-Oct3/4-TgDS: reverse, TAG CCA GGT TCG AGA ATC CA. Exo-Sox2 pMXs-Sox2-TgDS: reverse, GGT TCT CCT GGG CCA TCT TA. Exo-Klf4 pMXs-Klf4-TgDS: reverse, GGG AAG TCG CTT CAT GTG AG. Exo-c-Myc pMXs-c-Myc-TgDS: reverse, AGC AGC TCG AAT TTC TTC CA. Partly dissociated iPSCs were re-suspended in embryoid body medium (DMEM/F12 medium, 1× N2 supplement and 1% FBS) and cultured on shaker (95 r.p.m.) at 37 °C. Medium was changed every 3–4 days. After 20 days, total RNA of embryoid bodies was extracted for further gene expression analyses by qPCR. All tissue culture samples were routinely tested for mycoplasma by PCR. One millilitre of media supernatants (with no antibiotics or fungizone) was collected for all cell lines, spun down and resuspended in TE buffer. Ten microlitres of each sample were used in PCR reaction with the following primers: forward, GGC GAA TGG GTG AGT AAC; reverse, CGG ATA ACG CTT GCG ACC T. Any positive sample was immediately discarded. Three hundred nanograms of total extracted RNA from each sample were subjected to microarray by using the Affymatrix GeneChip one-cycle target labelling kit (Affymatrix, Santa Clara, California) according to the manufacturer’s recommended protocols. The resultant biotinylated cRNA was fragmented and then hybridized to the GeneChip Human 1.0 ST Array (764,885 probes, 28,869 genes, 19,734 gene-level probe sets with putative full-length transcript support (GenBank and RefSeq)) on the basis of human genome, Hg18. Arrays were prepared at the University of California DNA Core Facility. Arrays were analysed by the Affy (Affymetrix pre-processing)29 Bioconductor software package for microarray data. Data were then normalized by the RMA (robust multichip averaging) method to background-corrected and normalized probe levels to obtain a summary expression of normalized values for each probe set. Normalized microarray samples were then clustered by a hierarchical approach based on a matrix of distances. Normalized expression data were used to create a distance matrix that was calculated on the basis of Euclidean distance between the transcripts over a pair of samples representing a variation between two samples. Having the distances for all pairs of samples, a linkage method is used to cluster samples in a dendrogram by using calculated distances (sample expression similarities). This method also creates a heat map to graphically show the expression correlation between the samples. RNA samples were reverse transcribed into cDNA using the Super Script III First Strand Synthesis System (Invitrogen, California) according to the manufacturer’s instructions. Reactions were run on the Bio-Rad detection system using Sybr-green master mix (Bio-Rad). Primers were selected from Primerbank; validated database (http://pga.mgh.harvard.edu/primerbank/) and specificities were confirmed by melting curve analysis through a Bio-Rad detection system. Sequences of the primers are described in Supplementary Table 12. Quantitative analysis used the comparative threshold cycle method30. GAPDH was used as housekeeping gene. Each sample was run in triplicate. The RNA-seq analyses were previously described by our group31. Briefly, RNAs were isolated using the RNeasy Mini kit (Qiagen). A total of 1,000 ng of RNA was used for library preparation using the Illumina TruSeq RNA Sample Preparation Kit. The RNAs were sequenced on Illumina HiSeq2000 with 50 bp paired-end reads, generating 50 million high-quality sequencing fragments per sample on average. For validation purposes of biological samples subjected to RNA-seq, hESC and iPSC data available from the literature were downloaded and used to compare with our sequenced cell lines. The two hESC lines used are available (HUES-6, referred as ES(HUES), SRR873630, http://www.ncbi.nlm.nih.gov/sra/SRX290739; and H1, referred to here as ES(H1), SRR873631, http://www.ncbi.nlm.nih.gov/sra/SRX290740). The two human iPSC lines used are available under accession codes SRR873619 (referred to here as iPS(TD,1)) and SRR873620 (referred to here as iPS(TD,2)). RNA-seq enrichment used WebGestalt32 and Cytoscape33 software plugins, considering only categories having statistical significance (P < 0.05). Genes tested for differential expression were used as the background for GO annotation and enrichment analysis. NPCs were seeded onto poly-ornithine/laminin-coated six-well plates at a total number of 105 cells per well on day 0. Medium change was done on day 2. Cells were collected and counted on day 4. NPCs were resuspended, dissociated with accutase and fixed using fixation buffer (BioLegend) for 15 min followed by three PBS washes. The cell pellet was incubated and kept in Perm III buffer (BD Biosciences) at −20 °C until needed for the experiment. A total of 106 cells were incubated with antibodies Sox1 (PE), Sox2 (APC) or Nestin (PE) and Pax6 (APC) (Bd Biosciences) for 30 min and then washed three times before being resuspended for cell analyses. Cells were analysed in a plate reader mode using FACS Canto II machine (BD Biosciences). Cells were fixed in 4% paraformaldehyde for 10–20 min, washed with PBS three times (5 min each), permeabilized with 0.1% Triton X-100 for 15 min, incubated in blocking solution (2% BSA) for 1 h at room temperature and then in primary antibodies (goat anti-Nanog, Abcam ab77095, 1:500; rabbit anti-Lin28, Abcam ab46020, 1:500; rabbit anti-Oct4, Abcam ab19857, 1:500; mouse anti-SSEA4, Abcam ab16287, 1:200; mouse anti-Nestin, Abcam ab22035, 1:200; rabbit anti-Musashi1, Abcam ab52865, 1:250; rat anti-CTIP2, Abcam ab18465, 1:250; rabbit anti-SATB2, Abcam ab34735, 1:200; chicken anti-MAP2, Abcam ab5392, 1:1,000; rabbit anti-FZD9, Origene TA314730, 1:150; chicken anti-EGFP, Abcam ab13970, 1:1,000; rabbit anti-Synapsin1, EMD-Millipore AB1543P, 1:500; mouse anti-Vglut1, Synaptic Systems 135311, 1:500; rabbit anti-Homer1, Synaptic Systems 160003, 1:500) overnight at 4 °C. The next day, cells were washed with PBS three times (5 min each), incubated with secondary antibodies (Alexa Fluor 488, 555 and 647, Life Technologies, 1:1,000) for 1 h at room temperature and washed with PBS three times (5 min each). Nuclei were stained using DAPI (1:10,000). Slides or coverslips were mounted using ProLong Gold antifade mountant (Life Technologies). One million NPCs were harvested to single-cell suspension in 1mL PBS, then fixed by addition of 3 mL of 100% ethanol and stored at 4 °C for at least 2 h. NPC pellets were washed once with 5 mL PBS. After removal of PBS, cells were resuspended in 1 mL of propidium iodide (PI) staining solution (0.1% (v/v) Triton X-100, 10 μg/mL PI and 100 μg/mL RNase A in 1× PBS). WS and TD NPC samples were analysed by FACS on a Becton Dickinson LSRI, and gating of subG1 population (cells with fragmented DNA) was examined using FlowJo flow cytometry analysis software. Caspase activity was measured using a Green FLICA Caspases 3 & 7 Assay Kit (ImmunoChemistry Technologies). Briefly, NPCs were harvested, washed and stained with 1× carboxyfluorescein Fluorochrome Inhibitor of Caspase Assay (FAM-FLICA) reagent, 10 μg/mL Hoechst and 10 μg/mL propidium iodide (PI). Samples were analysed on the NC-3000 using the pre-optimized Caspase Assay. The population with caspase activity was used to analyse for apoptosis. NPC proliferation was assessed using BD Pharmingen BrdU Flow Kits (BD Biosciences) according to the manufacturer’s protocol. Briefly, NPCs were incubated with 1 μM BrdU for 45 min at 37 °C and harvested to single-cell suspension. NPCs were then fixed and permeabilized using BD Cytofix/Cytoperm Buffer and stained using FITC-conjugated anti-BrdU antibody and 7-aminoactinomycin D (7-AAD), a fluorescent dye for labelling DNA. Fluorescence-activated cell sorting (FACS) was done on LSRFortessa (BD Biosciences) and, to obtain the percentage of the BrdU-positive population, the cell-cycle profiles were analysed using FlowJo flow cytometry analysis software. Commercially available lentiviral vectors (pLKO.1) expressing short hairpin RNAs (shRNAs) against FZD9 under the control of the U6 promoter (Thermo Scientific) were engineered to express the Discosoma sp. red fluorescent protein (RFP) mCherry under the control of the hPGK (human phosphoglycerate kinase) promoter. The following shRNAs against FZD9 and a non-silencing scrambled control shRNA were selected (Thermo Scientific): shRNA-control, 5′-TTC TCC GAA CGT GTC ACG T-3′; shRNA-FZD9, 5′-ATC TTG CGG ATG TGG AAG AGG-3′. For rescue experiments, FZD9 cDNA was amplified from TD NPC cDNA as template by the following primer pair: 5′-CCG AGA TCT TCG AGG TGT GTG GGG TTC TCC AAA G-3′; 5′-TCT AGA GCC ACC ATG GCC GTA GCG CCT CTG-3′. The reaction was performed using Phusion High-Fidelity DNA polymerase (New England Biolabs) according to the manufacturer’s protocol. The FZD9 cDNA was cloned into a lentiviral vector driven by the ubiquitin promoter followed by a self-clevage peptide and GFP sequence. The specificity and efficiency of shRNA-control, shRNA-FZD9, and the FZD9-WT constructs were verified by co-transfection into HEK-293 cells. Cell lysates were collected and analysed by western blot analysis with anti-FZD9 antibodies (Aviva OAEC02415, 1:1,000). CHIR-98014 (Selleckchem) was resuspended according to manufacturer’s instructions into 10 mM stock using DMSO and then diluted to 100 μM. Final concentration used in cells was 100 nM of CHIR-98014, whereas the vehicle cells received only DMSO. For qPCR experiments, NPCs were propagated in six-well plates until 70% confluency and then treated with CHIR-98014 for 6 h to have their RNA collected using Trizol as previously described. For the NPC counting experiment, cells were seeded in six-well plates as described in the presence of CHIR-98014 or DMSO, in triplicates (TD and WS). After 48 h, the culture medium was changed and treatment was repeated. Cells were collected and counted after 96 h of incubation. The TD NPCs were lifted into suspension and maintained on a shaker (95 r.p.m.) to form neurospheres for 3 weeks. For the first week, the spheres were grown with N2B27 medium. The neurospheres were overlaid with the astrocyte medium (Lonza) for the remaining 2 weeks. The neurospheres were plated onto poly-ornithine- and laminin-coated plates and expanded for two to three passages before experimentation. Co-cultures of neurons and astrocytes were prepared for morphometric and functional analyses. NPCs were lysed in RIPA buffer with protease inhibitor. Rabbit anti-FZD9 antibody (Aviva OAEC02415, 1:1,000) and mouse anti-β-actin (Abcam ab8226, 1:3,000) were used as primary antibodies. IRDye 800CW goat anti-rabbit and IRDye 680RD goat anti-mouse (1:10,000) were used as secondary antibodies. The Odyssey system was used for signal detection. Signal intensities were measured using the Odyssey Image Studio and semi- quantitative analysis of FZD9 signal intensity was corrected with respect to β-actin relative quantification. A paired t-test analysis with P < 0.05 was used in the comparison of TD and WS FZD9 signal intensity normalized data. Co-localized Vglut (presynaptic) and Homer1 (postsynaptic) puncta were quantified after three-dimensional reconstruction of z-stack random images for all individuals and from two different experiments. Slides were analysed under a fluorescence microscope (Z1 Axio Observer Apotome, Zeiss). Only puncta in proximity of MAP2-positive processes were scored. Specific target amplification was performed in individual dissociated NPCs or 6-week-old neurons using C1 Single-Cell and BioMark HD Systems (Fluidigm), according to the manufacturer’s protocol and as described previously34, 35, 36. Briefly, single cells were captured on a C1 chip (10- to 17-μm cells) and cell viability was checked using a LIVE/DEAD Cell Viability/Cytotoxicity kit (Life Technologies). After lysis, RNA was reverse transcribed into cDNA with validated amplicon-specific DELTAgene Assays (Supplementary Table 13) using SuperScript III RT Platinum Taq Mix. Specific target amplification was performed by 18 cycles of 95 °C denaturation for 15 s and 60 °C annealing and amplification for 4 min. Each preamplified cDNA was mixed with 2× SsoFast EvaGreen Supermix with Low ROX (Bio-Rad) and then pipetted into an individual sample inlet in a 96.96 Dynamic Array IFC chip (Fluidigm). DELTAgene primer pairs (Supplementary Table 13) were diluted and pipetted into individual assay inlets in the same 96.96 Dynamic Array IFC chip. Quantitative PCR results were analysed using Fluidigm’s Real-time PCR Analysis software using the linear (derivative) baseline correction method and the automatic (gene) C threshold method with 0.65 curve quality threshold. Hierarchical clustering heat map, PCA analyses, violin plots of log (expression of C values) (limit of detection = 24) and ANOVA statistical analysis were performed using Singular Analysis Toolset 3.0 (Fluidigm). Neuronal networks derived from human iPSCs were transduced with lentivirus carrying the Syn::RFP reporter construct. Cell cultures were washed with Krebs HEPES buffer (KHB) (10 mM HEPES, 4.2 mM NaHCO , 10 mM dextrose, 1.18 mM MgSO , 1.18 mM KH PO 4.69 mM KCl, 118 mM NaCl, 1.29 mM NaCl ; pH 7.3) and incubated with 2–5 μM Fluo-4AM (Molecular Probes/Invitrogen, Carlsbad, California) in KHB for 40 min. Five thousand frames were acquired at 28 Hz with a region of 256 pixels × 256 pixels (×100 magnification), using a Hamamatsu ORCA-ER digital camera (Hamamatsu Photonics K.K., Japan) with a 488 nm (FITC) filter on an Olympus IX81 inverted fluorescence confocal microscope (Olympus Optical, Japan). Images were acquired with MetaMorph 7.7 (MDS Analytical Technologies, Sunnyvale, California), processed and analysed using individual circular regions of interest (ROI) on ImageJ and Matlab 7.2 (Mathworks, Natick, Massachusetts). Syn::RFP+ neurons were selected after confirmation that calcium transients were blocked with 1 mM of tetrodotoxin (TTX). The amplitude of signals was presented as relative fluorescence changes (ΔF/F) after background subtraction. The threshold for calcium spikes was set at the 95th percentile of the amplitude of all detected events. For whole-cell patch-clamp recordings, individual coverslips containing live 1-month-old neurons were transferred into a heated recording chamber and continuously perfused (1 mL/min) with artificial cerebrospinal fluid bubbled with a mixture of CO (5%) and O (95%) and maintained at 25 °C. Artificial cerebrospinal fluid contained (in mM) 121 NaCl, 4.2 KCl, 1.1 CaCl2, 1 MgSO , 29 NaHCO , 0.45 NaH PO -H O, 0.5 Na HPO and 20 glucose (all chemicals from Sigma). Whole-cell recordings were performed using a digidata 1440A/ Multiclamp 700B and Clampex 10.3 (Molecular devices). Patch electrodes were filled with internal solutions containing 130 mM K-gluconate, 6 mM KCl, 4 mM NaCl, 10 mM Na-HEPES, 0.2 mM K-EGTA; 0.3 mM GTP, 2 mM Mg-ATP, 0.2 mM cAMP, 10 mM d-glucose, 0.15% biocytin and 0.06% rhodamine. The pH and osmolarity were adjusted for physiological conditions. Data were all corrected for liquid junction potentials, electrode capacitances were compensated on-line in cell-attached mode and a low-pass filter at 2 kHz was used. The access resistance of the cells in our sample was around 37 MΩ with resistance of the patch pipettes 3–5 MΩ. Spontaneous synaptic AMPA events were recorded at the reversal potential of Cl− and could be reversibly blocked by AMPA receptor antagonist (10 μM NBQX, Sigma). Spontaneous synaptic GABA events were recorded at the reversal potential of Na+ and could be reversibly blocked with GABA receptor antagonist (10 μM SR95531, Sigma). Using 12-well MEA plates from Axion Biosystems, we plated the same density of NPCs from TD and WS individuals in triplicate. Each well was seeded with 10,000 NPCs that were induced into neuronal differentiation as previously described. Each well was coated with poly-l-ornithine and laminin before cell seeding. Cells were fed once a week and measurements were taken before the medium was changed. Recordings were performed using a Maestro MEA system and AxIS software (Axion Biosystems), using a band-pass filter with 10 Hz and 2.5 kHz cutoff frequencies. Spike detection was performed using an adaptive threshold set to 5.5 times the standard deviation of the estimated noise on each electrode. Each plate first rested for 5 min in the Maestro, and then 5–10 min of data were recorded to calculate the spike rate per well. MEA analysis was performed using the Axion Biosystems Neural Metrics Tool, wherein electrodes that detected at least five spikes per minute were classified as active electrodes. Bursts were identified in the data recorded from each individual electrode using an adaptive Poisson surprise algorithm. Network bursts were identified for each well, using a non-adaptive algorithm requiring a minimum of ten spikes with a maximum inter-spike interval of 100 ms. Only channels that exhibited bursting activity (more than ten spikes in 5 min interval) were included in this analysis. After measurement, neurons were immunostained to check morphology and density. We used six post-mortem brains (two WS and four TD) that were gender-, age- and hemisphere-matched. All brain specimens were harvested within a post-mortem interval of 18–30 h and had been immersed and fixed in 10% formalin for up to 20 years. For the purpose of the present experiments, samples were obtained from anatomically well-identified cortical areas in a consistent manner across specimens. Tissue blocks approximately 5 mm3 were removed from primary somatosensory cortex (Brodmann area 3) and primary motor cortex (Brodmann area 4) in the arm/hand knob region of the pre- and postcentral gyri, respectively, and from the secondary visual area (Brodmann area 18) from approximately 1.4 cm dorsally to the occipital pole and 2 cm from the midline37, 38. We focused specifically on these parts of the cortex because pathologies in dendritic morphology in these areas have been reported in other neurodevelopmental disorders39, 40, 41. In addition, pyramidal neurons in the selected areas reach their mature-like morphology early in development and start displaying dendritic pathologies sooner than high integration areas, such as the prefrontal cortex, allowing comparison of post-mortem findings with iPSC-derived neurons in early stages of development42, 43. Sampled tissue blocks were processed using an adaptation of the Golgi–Kopsch method44, which has been shown to give good results with tissue that has been fixed for long periods45. Briefly, blocks were immersed in a solution of 3% potassium dichromate, 0.5% formalin for 8 days, followed by immersion into 0.75% silver nitrate for 2 days. Blocks were then sectioned on a vibratome, perpendicular to the pial surface, at a thickness of 120 μm. Golgi sections were cut into 100% ethyl alcohol and transferred briefly into methyl salicylate followed by toluene, mounted onto glass slides and coverslipped. Adjacent blocks from each region were sectioned at 60 μm and stained with thionin for visualization of cell bodies and laminar organization, which enabled identification of the position of each neuron within a specific cortical layer. Cytoarchitectonic analysis of histological sections from each block confirmed that tissue was sampled from the ROI and that the Golgi-impregnated pyramidal neurons were located in cortical layers V/VI. Cortical neurons from all six post-mortem brains were used in the study. Neurons included in the morphological analysis did not display degenerative changes46. Only neurons with fully impregnated soma, apical dendrites with present oblique branches and at least two basal dendrites with third-order segments were chosen for the analysis47. To minimize the effects of cutting on dendritic measurements, we included neurons with cell bodies located near the centre of 120-μm-thick histological sections, with natural terminations of higher-order dendritic branches present where possible37, 47. Inclusion of the neurons completely contained within 120-μm sections biases the sample towards smaller neurons, leading to the underestimation of dendritic length48; therefore, we applied the same criteria blinded across all WS and TD specimens, and we thus included the neurons with incomplete endings if they were judged to otherwise fulfil the criteria for successful Golgi impregnation. All neurons were oriented with apical dendrite perpendicular to the pial surface; inverted pyramidal cells as well as magnopyramidal neurons were excluded from the analysis. Neuronal morphology was quantified along x-, y-, and z-coordinates using Neurolucida version 10 software (MBF Bioscience, Williston, Vermont) connected to a Nikon Eclipse 80i microscope, with a ×40 (0.75 numerical aperture) Plan Fluor dry objective. Tracings were conducted on both apical and basal dendrites, and the results reflect summed values for both types of dendrite per neuron. Following the recommendation that the applications of Sholl’s concentric spheres or Eayrs’ concentric circles for the analysis of neuronal morphology are not adequate when neuronal morphology is analysed in three dimensions48, we conducted dendritic tree analysis with the following measurements37, 47: (1) soma area—cross sectional surface area of the cell body; (2) dendritic length—summed total length of all dendrites per neuron; (3) dendrite number—number of dendritic trees emerging directly from the soma per neuron; (4) dendritic segment number—total number of segments per neuron; (5) dendritic spine/protrusion number—total number of dendritic spines per neuron; (6) dendritic spine/protrusion density—average number of spines per 20 μm of dendritic length; and (7) branching point number—number of nodes (points at the dendrite where a dendrite branches into two or more) per neuron. Dendritic segments were defined as parts of the dendrites between two branching points—between the soma and the first branching point in the case of first-order dendritic segments, and between the last branching point and the termination of the dendrite in the case of terminal dendritic segments. Since the long formalin-fixation time could have resulted in degradation of dendritic spines, spine values might be underestimated and are thus reported here with caution. All of the tracings were accomplished blind to brain region and diagnostic status. The iPSC-derived sample consisted of EGFP-positive 8-week-old neurons with pyramidal- or ovoid-shaped soma and at least two branched neurites (dendrites) with visible spines/protrusions. Protrusions from dendritic shaft, which morphologically resembled dendritic spines in post-mortem specimens, were considered and quantified as dendritic spines in iPSC-derived neurons. The neurites were considered dendrites on the basis of the criteria applied in post-mortem studies: (1) thickness that decreased with the distance from the cell body; (2) branches emerging under acute angle; and (3) presence of dendritic spines. In addition, only enhanced-GFP-positive neurons with nuclei co-stained with CTIP2, indicative of layer V/VI neurons, and with the dendrites displaying evenly distributed fluorescent stain along their entire length, were included in the analysis. The morphology of the neurons was quantified along x-, y-, and z-coordinates using Neurolucida version 9 software (MBF Bioscience, Williston, VT) connected to a Nikon Eclipse E600 microscope with a ×40 oil objective. No distinction was made between apical and basal dendrites, and the results reflect summed length values of all neurites/dendrites per neuron, consistent with what was done for the post-mortem neurons. The same set of measurements used in the analysis of Golgi-impregnated neurons was applied to the analysis of iPSC-derived neurons, and all of the tracings were accomplished blind to the diagnostic status and were conducted by the same rater (B.H.-M.). Intra-rater reliability was assessed by having the rater trace the same neuron after a period of time. The average coefficient of variation between the results of retraced neurons was 2% for soma area (SA), total dendritic length (TDL), dendritic segment number (DSN) and branching point number (BPN), and 3% for dendritic spine/protrusion number (DPN); there was no variation in tree/dendrite number (TN) in different tracings of the same neuron. The accuracy was further checked by having three individuals (B.H.-M., B.J. and L.S.) trace the same neuron. MRI scanning was completed in 19 participants with WS (aged 19–43 years; mean 29.0, s.d. 8.8; 11 males, 8 females) and 19 TD comparison participants (aged 16–43 years; mean 26.2, s.d. 7.3; 8 males, 11 females). There was no significant difference between the groups in age (t = 1.0, P < 0.30) or in gender ratio (Pearson’s χ2 = 0.95, P < 0.33). A standardized multiple modality high-resolution structural MRI protocol was implemented, involving three-dimensional T - and T -weighted volumes and a set of diffusion-weighted scans. Imaging data were obtained at the University of California San Diego Radiology Imaging Laboratory on a 1.5 T GE Signa HDx 14.0M5 TwinSpeed system (GE Healthcare, Waukesha, Wisconsin) using an eight-channel phased array head coil. A three-dimensional inversion recovery spoiled gradient echo (IR-SPGR) T -weighted volume was acquired with pulse sequence parameters optimized for maximum grey/white matter contrast (echo time = 3.9 ms, repetition time = 8.7 ms, inversion time = 270 ms, flip angle = 8°, difference in echo times = 750 ms, bandwidth = ± 15.63 kHz, field of view = 24 cm, matrix = 192 × 192, voxel size = 1.25 mm × 1.25 mm × 1.2 mm). All MRI data were collected using prospective motion (PROMO) correction for non-diffusion imaging49. This method has been shown to improve image quality, reduce motion-related artefacts, increase the reliability of quantitative measures and improve the clinical diagnostic utility of MRI data obtained in children and clinical groups50, 51. Standardized quality control procedures were followed for both raw and processed data, including visual inspection ratings by a trained imaging technician and computer algorithms testing general image characteristics as well as aspects specific to each imaging modality, such as contrast properties, registrations and artefacts from motion and other sources. Participants included in the current analyses were only those who passed all raw and processed quality control measures. Image post-processing and analysis were performed using FreeSurfer software suite (http://surfer.nmr.mgh.harvard.edu/). Surface-based cortical reconstruction and subcortical volumetric segmentation procedures have been shown elsewhere52, 53, 54, 55, 56, 57, 58. Briefly, a three-dimensional model of the cortical surface was generated using MRI scans with four attributes: white matter segmentation; tessellation of the grey/white matter boundary; inflation of the folded, tessellated surface; and correction of topological defects53, 54. Cortical thickness was measured using the distances from each point on the white matter surface to the pial surface57. Cortical surface area was measured at the pial surface for the entire cerebrum and for each parcel of the Desikan and Destrieux atlases53, 54, 58, 59. Means ± s.e.m. for each parameter were obtained from samples described in Supplementary Table 1. There were no statistical methods used to predetermine sample size. The experiments were not randomized. All of the tracings were accomplished blind to brain region and diagnostic status. All statistical analyses were done using Prism (Graphpad). Before statistical analysis comparing means between three to five unmatched groups of data, normal distribution was tested using D’Agostino and Pearson omnibus normality test and variance similarity was tested using Bartlett’s test for equal variances. Means of three to five unmatched groups, where normal distribution and equal variances between groups were confirmed, were statistically compared using one-way ANOVA and Tukey’s post hoc test. Otherwise, a Kruskal–Wallis test and Dunn’s multiple comparison test were used. Before statistical analysis comparing means between two unmatched groups of data, normal distribution was tested using D’Agostino and Pearson omnibus normality test and variance similarity was tested using an F test to compare variances. To compare the means of two groups where normal distribution and similar variance between groups were confirmed, Student’s t test was used. Otherwise, a Mann–Whitney test was used. Significance was defined as *P < 0.05, **P < 0.01, ***P < 0.001 or ****P < 0.0001.
News Article | November 10, 2016
Project led By University of Maryland School of Medicine will focus on accelerating the use of vaccines to protect from disease that kills more than 220,000 annually Baltimore, MD, November 10, 2016 - Typhoid fever, a bacterial infection that causes high fever and other disabling symptoms, remains a serious global problem in the developing world: it kills almost a quarter of a million people annually, and infects about 21 million. To help speed the introduction of, and access to, new and more effective typhoid vaccines, the University of Maryland School of Medicine (UM SOM) Center for Vaccine Development (CVD) has received a grant of $36.9 million from the Bill & Melinda Gates Foundation. The project, known as Typhoid Vaccine Acceleration Consortium (TyVAC), is a partnership with the Oxford Vaccine Group at the University of Oxford and PATH, an international nonprofit global health organization based in Seattle. TyVAC will focus on conjugate vaccines, which can trigger a stronger immune response in certain vulnerable populations, such as infants and children, than current typhoid vaccines. TyVAC will employ a multidisciplinary approach to study and control typhoid, and generate evidence that informs global policies. The project will work closely with governments and policymakers to introduce vaccines in lower-income countries with a high burden of typhoid. The effort will also examine how well the vaccine rollouts work in early adopter countries. The project's overall goal is to support accelerated, evidence-based decisions for new typhoid conjugate vaccine introductions that will significantly reduce the severe health and economic burdens of the disease. "Typhoid fever disproportionately impacts children and poor populations," said Kathleen Neuzil, MD, MPH, FIDSA, professor of medicine at UM SOM, director of CVD, and deputy director of the Institute for Global Health (IGH). "With our long history of work in typhoid and typhoid vaccines, we look forward to working with partners to catalyze action against this significant public health problem." "It is unconscionable that children are still dying by the thousands every year from diseases like typhoid that are completely preventable," said Anita Zaidi, director of the Enteric and Diarrheal Diseases team at the Bill & Melinda Gates Foundation. "The prevention and control of typhoid should be a global health priority and we are pleased to support the Typhoid Vaccine Acceleration Consortium as part of our overall strategy to combat typhoid through an integrated approach including access to clean water, improved sanitation, and immunization." At present, the currently available vaccines for typhoid fever are underutilized despite the substantial disease burden and a World Health Organization recommendation for the use of typhoid vaccines in areas of high burden. Typhoid conjugate vaccines have the promise to overcome some of the barriers of the currently available vaccines, providing a stronger immune response, a longer duration of protection, and the ability to be incorporated into the routine vaccination schedule targeted at children less than two years old. "We are excited to work in partnership with CVD to bring our expertise on typhoid infections and vaccines to the consortium and improve health through TyVAC," said Andrew Pollard, MD, PhD, professor of pediatric infection and immunity at the University of Oxford, and director of the Oxford Vaccine Group. Population density, limited sanitation, and poor water quality can provide a breeding ground for typhoid. "With increasing urbanization, we could see an even greater burden of typhoid," noted Deborah Atherly, PhD, head of Policy, Access, and Introduction for PATH's Center for Vaccine Innovation and Access. "Through TyVAC, we will work to ensure that typhoid vaccines finally reach those who need them most." "Typhoid is a significant public health problem in many parts of the world," said UM SOM Dean E. Albert Reece, MD, PhD, MBA, who is also vice president for medical affairs at the University of Maryland and the John Z. and Akiko K. Bowers Distinguished Professor. "Over several decades, the CVD has helped save millions of lives. This generous grant from the Gates Foundation will allow our scientists, working with national and international partners, to continue with this crucial work." The CVD at the University of Maryland School of Medicine works nationally and internationally to prevent disease and save lives through the development and delivery of vaccines. As an academic research center, CVD is engaged in the full range of vaccinology, including basic science research, vaccine development, pre-clinical and clinical evaluation, and post-marketing field studies. Learn more at http://medschool. . The University of Maryland School of Medicine was chartered in 1807 and is the first public medical school in the United States and continues today as an innovative leader in accelerating innovation and discovery in medicine. The School of Medicine is the founding school of the University of Maryland and is an integral part of the 11-campus University System of Maryland. Located on the University of Maryland's Baltimore campus, the School of Medicine works closely with the University of Maryland Medical Center and Medical System to provide a research-intensive, academic and clinically based education. With 43 academic departments, centers and institutes and a faculty of more than 3,000 physicians and research scientists plus more than $400 million in extramural funding, the School is regarded as one of the leading biomedical research institutions in the U.S. with top-tier faculty and programs in cancer, brain science, surgery and transplantation, trauma and emergency medicine, vaccine development and human genomics, among other centers of excellence. The School is not only concerned with the health of the citizens of Maryland and the nation, but also has a global presence, with research and treatment facilities in more than 35 countries around the world. Learn more at http://medschool. . The University of Oxford is one of the top five higher education institutions in the world and hosts the Oxford Vaccine Group (OVG) in the Department of Paediatrics (http://www. ). OVG is a vaccine design, development, clinical trials, and laboratory evaluation research group with specific expertise in vaccine evaluation in paediatric populations. The University of Oxford has strategically made a major investment in infrastructure to support research on vaccines and immunity over the past two decades. The OVG is one of the largest academic research groups in the world focused on designing, developing, and evaluating vaccines for children, as well as characterizing immune response to vaccines and infectious diseases. PATH is the leader in global health innovation. An international nonprofit organization, we save lives and improve health, especially among women and children. We accelerate innovation across five platforms--vaccines, drugs, diagnostics, devices, and system and service innovations--that harness our entrepreneurial insight, scientific and public health expertise, and passion for health equity. By mobilizing partners around the world, we take innovation to scale, working alongside countries primarily in Africa and Asia to tackle their greatest health needs. Together, we deliver measurable results that disrupt the cycle of poor health. Learn more at http://www.
News Article | November 1, 2016
There is a substantial gap in knowledge of pharmacokinetic changes in pregnancy and their clinical consequences, according to a study published by Shinya Ito and colleagues from the Hospital for Sick Children, Toronto, Canada, in PLOS Medicine. Pregnant women take a variety of medications, including prescription and over-the-counter medications, with more than 90% of women in some areas reporting use of at least one medication during pregnancy. However, information may not be available on whether or how to adjust dosing to compensate for the physiologic changes that occur during pregnancy. The researchers conducted a systematic review of the biomedical literature, and identified 198 studies, involving 121 different medications, that measured changes in pharmacokinetics (absorption, distribution, metabolism, and excretion of drugs) during pregnancy and/or the resultant clinical impact of these changes. Across the different drug classes, they frequently observed a decrease in drug exposure mainly due to increased elimination. However, only a few studies described changes in clinical outcomes associated with altered pharmacokinetics during pregnancy. The authors say: "It is essential for clinicians to be aware of these unique pregnancy -related changes in pharmacokinetics, and to critically examine their potential clinical implications." The authors received no specific funding for this work. The authors have declared that no competing interests exist. Division of Clinical Pharmacology and Toxicology, Hospital for Sick Children, Toronto, Ontario, Canada Hospital Library, Hospital for Sick Children, Toronto, Ontario, Canada Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada Department of Paediatrics, University of Toronto, Toronto, Ontario, Canada Department of Pharmacology & Toxicology, University of Toronto, Toronto, Ontario, Canada Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, Ontario, Canada IN YOUR COVERAGE PLEASE USE THIS URL TO PROVIDE ACCESS TO THE FREELY AVAILABLE PAPER: http://journals. LCC is supported by a National Institute for Health Research Professorship, RP-2014-05-019. AD is supported by the National Institute for Health Research University College London Hospitals Biomedical Research Centre. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. AD is Director and cofounder of Magnus Growth, part of Magnus Life Science, a company that is aiming to take to market a novel treatment for fetal growth restriction. Chappell LC, David AL (2016) Improving the Pipeline for Developing and Testing Pharmacological Treatments in Pregnancy. PLoS Med 13(11): e1002161. doi:10.1371/journal.pmed.1002161 IN YOUR COVERAGE PLEASE USE THIS URL TO PROVIDE ACCESS TO THE FREELY AVAILABLE PAPER: http://journals.
News Article | November 8, 2016
Scientists from Children's Health Research Institute, a program of Lawson Health Research Institute, and Western University have developed a new blood test that identifies with greater than 90 per cent certainty whether or not an adolescent athlete has suffered a concussion. Diagnosis of a clinically significant concussion, or a mild traumatic brain injury, can be difficult as it currently relies on a combination of patient symptom assessment and clinician judgement. Equally problematic are the decisions to stop play or activities, or when patients who have suffered a concussion can safely return to normal activities without risking further injury. In the new study, researchers have demonstrated that a blood test can now accurately diagnose a concussion using a form of blood profiling known as metabolomics. Dr. Douglas Fraser, a physician in the Paediatric Critical Care Unit at Children's Hospital, London Health Sciences Centre and Lawson scientist, led the study with his co-investigator Mark Daley, a professor in the Departments of Computer Science, Biology and Statistics & Actuarial Sciences at Western University. In the relatively inexpensive test, blood is drawn from an individual that may have suffered a concussion as the result of a sudden blow to the head (or from transmitted forces from a sudden blow to the body) within 72 hours of the incident. The scientists measure a panel of metabolites -- small molecules that are the products of the body's metabolism -- in the blood to search for distinct patterns that indicate a concussion has occurred. "This novel approach, to use blood testing of metabolites as a diagnostic tool for concussions, was exploratory and we were extremely pleased with the robustness of our initial results," says Dr. Fraser, also an Associate Professor in Western's Departments of Paediatrics, Physiology & Pharmacology and Clinical Neurological Sciences at the Schulich School of Medicine & Dentistry. "We looked at a host of patterns and it appears that those who suffered a concussion have a very different pattern than those who have not had a concussion." This new method, fully funded by the Children's Health Foundation and conducted by the Western Concussion Study Group, is unique in that previous attempts have looked unsuccessfully for a single highly accurate protein biomarker that can distinguish concussed from non-concussed adolescent patients. In this latest successful attempt, the researchers took a different approach and investigated a full spectrum of 174 metabolites. "We looked at all of these metabolites in concussed male adolescent patients and in non-concussed male adolescent patients and it turns out that the spectrum is really different," explains Daley, who is also Western's Associate Vice-President (Research) and a principal investigator at Western's Brain & Mind Institute. "There is no one metabolite that we can put a finger on but when we looked at all of them, those profiles are different enough that we could easily distinguish concussed patients from non-concussed. In fact, with fine tuning we can now look at sets of as few as 20-40 specific metabolites and maintain the diagnostic accuracy level of the test over 90 per cent." Concussion is a major public health concern, often resulting in significant acute symptoms and in some individuals, long-term neurological dysfunction. "The discovery of a blood test that can aid in concussion diagnosis is very important," says Dr. Fraser. "With further research, we anticipate that our blood test will also aid clinicians in predicting concussion outcome, as well as aid rehabilitation after concussion." The findings were recently published in the international journal Metabolomics. The technology is subject to a patent application filed through WORLDiscoveries®, the joint technology transfer office of Lawson and Western. "This relatively quick and inexpensive blood test for concussion is by far the most accurate reported with tremendous potential for clinical management and commercialization," says Kirk Brown, Manager of Business Development for Lawson.
News Article | November 7, 2016
LONDON, ON - Scientists from Children's Health Research Institute, a program of Lawson Health Research Institute, and Western University have developed a new blood test that identifies with greater than 90 per cent certainty whether or not an adolescent athlete has suffered a concussion. Diagnosis of a clinically significant concussion, or a mild traumatic brain injury, can be difficult as it currently relies on a combination of patient symptom assessment and clinician judgement. Equally problematic are the decisions to stop play or activities, or when patients who have suffered a concussion can safely return to normal activities without risking further injury. In the new study, researchers have demonstrated that a blood test can now accurately diagnose a concussion using a form of blood profiling known as metabolomics. Dr. Douglas Fraser, a physician in the Paediatric Critical Care Unit at Children's Hospital, London Health Sciences Centre and Lawson scientist, led the study with his co-investigator Mark Daley, a professor in the Departments of Computer Science, Biology and Statistics & Actuarial Sciences at Western University. In the relatively inexpensive test, blood is drawn from an individual that may have suffered a concussion as the result of a sudden blow to the head (or from transmitted forces from a sudden blow to the body) within 72 hours of the incident. The scientists measure a panel of metabolites - small molecules that are the products of the body's metabolism - in the blood to search for distinct patterns that indicate a concussion has occurred. "This novel approach, to use blood testing of metabolites as a diagnostic tool for concussions, was exploratory and we were extremely pleased with the robustness of our initial results," says Dr. Fraser, also an Associate Professor in Western's Departments of Paediatrics, Physiology & Pharmacology and Clinical Neurological Sciences at the Schulich School of Medicine & Dentistry. "We looked at a host of patterns and it appears that those who suffered a concussion have a very different pattern than those who have not had a concussion." This new method, fully funded by the Children's Health Foundation and conducted by the Western Concussion Study Group, is unique in that previous attempts have looked unsuccessfully for a single highly accurate protein biomarker that can distinguish concussed from non-concussed adolescent patients. In this latest successful attempt, the researchers took a different approach and investigated a full spectrum of 174 metabolites. "We looked at all of these metabolites in concussed male adolescent patients and in non-concussed male adolescent patients and it turns out that the spectrum is really different," explains Daley, who is also Western's Associate Vice-President (Research) and a principal investigator at Western's renowned Brain & Mind Institute. "There is no one metabolite that we can put a finger on but when we looked at all of them, those profiles are different enough that we could easily distinguish concussed patients from non-concussed. In fact, with fine tuning we can now look at sets of as few as 20-40 specific metabolites and maintain the diagnostic accuracy level of the test over 90 per cent." Concussion is a major public health concern, often resulting in significant acute symptoms and in some individuals, long-term neurological dysfunction. "The discovery of a blood test that can aid in concussion diagnosis is very important," says Dr. Fraser. "With further research, we anticipate that our blood test will also aid clinicians in predicting concussion outcome, as well as aid rehabilitation after concussion." The findings were recently published in the international journal Metabolomics. The technology is subject to a patent application filed through WORLDiscoveries®, the joint technology transfer office of Lawson and Western. "This relatively quick and inexpensive blood test for concussion is by far the most accurate reported with tremendous potential for clinical management and commercialization," says Kirk Brown, Manager of Business Development for Lawson. Western delivers an academic experience second to none. Since 1878, The Western Experience has combined academic excellence with life-long opportunities for intellectual, social and cultural growth in order to better serve our communities. Our research excellence expands knowledge and drives discovery with real-world application. Western attracts individuals with a broad worldview, seeking to study, influence and lead in the international community. As the research institute of London Health Sciences Centre and St. Joseph's Health Care London, and working in partnership with Western University, Lawson Health Research Institute is committed to furthering scientific knowledge to advance health care around the world. http://www. Children's Health Foundation is dedicated to raising and granting funds to support Children's Hospital at London Health Sciences Centre, Thames Valley Children's Centre and Children's Health Research Institute. Since 1922, funds raised have helped deliver exceptional care and support for children and their families by providing specialized paediatric care, equipment, education programs, therapy, rehabilitation services and research. Get to know how you can help save and improve kids' lives at http://www.
News Article | January 17, 2017
Popular baby chew toy Sophie, the Giraffe was reportedly found with mold growing inside it. The incident was reported first by paediatric dentist, Dana Chianese. Chew toys or a teether is a baby toy made generally out of rubber. It is used as a soothing tool for teething infants. Chianese told Goodhousekeeping that she used to recommend Sophie, the Giraffe, as a teething toy to parents but not anymore. What changed her mind you wonder? About a month ago Dana noticed musty smelling air coming out of the toy, which her children prefer to play with. "I decided to cut into Sophie out of curiosity and discovered a science experiment living inside. Smelly, ugly mold living in my infant's favorite chew toy!," shared Chianese with the publication. Chianese states that she cleans the toy strictly following the instructions given by the company, using soapy hot water and applying it on the toy with a damp sponge. Dana claims to have never put the toy in water completely for cleaning purposes. Allegations against Sophie, the Giraffe, is not an isolated incident. Stephanie Opera, a concerned mother, also reviewed the product on Amazon and tried to warn other parents about the teething toy through her review. "Beware!! If you have a drooly baby, moisture will get in the hole and you'll end up with mold! We've had ours for two years and the entire inside is coated with black mold!" wrote Stephanie on Amazon. A What to Expect user, also found mold inside the toy. On hearing news that the toy's legs get caught in babies' throat, the user consulted with her DH and decided to chop off the legs. It could be safely assumed that she was not expecting to see mold inside the toy. While parents are panicking over the fact that mold has been found inside a teether frequently used by infants, Dr. Luba Konopasek, an Assistant Professor of Paediatrics at New York Presbyterian and Weil Cornell Medical Center asks parents not to get too alarmed. Dr. Konopasek told care.com that parents need not get too anxious about the mold situation, until and unless their child has an immune disorder like mold allergy. In that case it is safe to say, keep the child away from Sophie. Carolyn Forte, director of Cleaning Lab suggests that parents clean and thoroughly dry the plastic toys to prevent the growth of mold inside them. Laurie Schraenen, a spokeswoman for Sophie the Giraffe recently issued a statement on being contacted by Good Housekeeping. In the statement, Schraenen clearly stated that Sophie the Giraffe was made out of 100% natural rubber and the company recommends cleaning the toy, strictly abiding by the instructions given by the company. The company has promised it will look into the matter seriously and will conduct further examination to reach to the bottom of the matter. © 2017 Tech Times, All rights reserved. Do not reproduce without permission.
News Article | December 1, 2016
CALGARY, AB--(Marketwired - December 01, 2016) - Sudden infant death syndrome (SIDS) is the sudden, unexplainable death of children under the age of one. While rates have significantly decreased due to research and public advocacy, it remains the leading cause of infant death in Canada. Researchers from the University of Calgary's Cumming School of Medicine are one step closer to understanding how exposure to cigarette smoke increases the risk of SIDS, and how the cumulative effect of high environmental temperatures (caused by factors such as co-bedding and over-wrapping) and exposure to infection also increases the risk. The study findings were recently published in the journal American Journal of Physiology - Regulatory, Integrative and Comparative Physiology. "It's important to understand how these environmental factors increase risk so we can better alert parents of the potential danger of these situations," says Dr. Shabih Hasan, Professor, Department of Paediatrics, member of the university's Alberta Children's Hospital Research Institute and senior author on the study. The study exposed animal models to cigarette smoke in utero and subsequently exposed them to infection-causing bacteria and high environmental temperatures when they were born. The study found that the smoke-exposure group had increased temporary interruptions of breathing, higher rates of cytokine (inflammation causing messenger cells from the immune system) production and an increased heart rate when exposed to infection and high environmental temperature. Researchers also observed that in both the smoking and non-smoking group, increased body temperature and exposure to infection suppressed the heart rate to levels normally only observed in low oxygen environments (experienced in scenarios such as sleeping under a blanket). While the studies were conducted in animal models, Hasan says these findings are relevant to humans as they are modelled after evidence in human infants who died from SIDS. Currently, one in 2,000 babies in Canada will die from SIDS. "These findings emphasize the importance of advocating for a safe sleep environment for babies," says Hasan. "This includes avoidance of over-wrapping infants, setting a reasonable room temperature, avoidance of co-bedding, placing babies to sleep on their back and the avoidance of cigarette smoke both during pregnancy and after infants' birth." Sarah Cormier is an Airdrie mother whose daughter, Quinn Isla, died of SIDS at the age of four months in 2014. "Our initial reaction was of shock, loneliness and surprise," she says. "SIDS is every parent's worst nightmare and we are now living it. We were aware of the risk factors but it's hard to believe that it still happens and that it happened to us." Cormier says Hasan's research and any other research investigating the mysteries behind SIDS is welcomed. "We are hopeful that this is one step closer to finding out what happens to these babies," she says. Since the 1990s, "Back to Sleep" public advocacy and media campaigns decreased the SIDS rate by more than 50 per cent. However, the rates of SIDS must be further decreased as one infant death is too many. Hasan and his team are certain that more informed infant care practices will further reduce the SIDS rates. Important changes in infant care practice involve the following: About the University of Calgary The University of Calgary is making tremendous progress on its journey to become one of Canada's top five research universities, where research and innovative teaching go hand in hand, and where we fully engage the communities we both serve and lead. This strategy is called Eyes High, inspired by the university's Gaelic motto, which translates as 'I will lift up my eyes.' For more information, visit ucalgary.ca. Stay up to date with University of Calgary news headlines on Twitter @UCalgary. For details on faculties and how to reach experts go to our media centre at ucalgary.ca/mediacentre. About the Cumming School of Medicine The University of Calgary's Cumming School of Medicine is a leader in health research, with an international reputation for excellence and innovation in health care research and education. On June 17, 2014, the University Of Calgary Faculty Of Medicine was formally named the Cumming School of Medicine in recognition of Geoffrey Cumming's generous gift to the university. For more information, visit cumming.ucalgary.ca, or follow us on Twitter @UCalgaryMed. About the Alberta Children's Hospital Research Institute The Alberta Children's Hospital Research Institute (ACHRI) is a multi-disciplinary institute of the University of Calgary, Alberta Health Services and the Alberta Children's Hospital Foundation. Membership encompasses the faculties of arts, education, kinesiology, medicine, nursing, science, social work and veterinary medicine. ACHRI co-ordinates child and maternal health research from bench to bedside with a vision of giving mothers and children the best health care possible. Working together with an incredible breadth of expertise, the institute members are determined to find the causes of disease, advance medical treatments and prevent illness and injury in children.
News Article | December 19, 2016
EMBU, Kenya--(BUSINESS WIRE)--Camusat, a market leader in implementing telecom infrastructures, equipped Kyeni Consolata Hospital in Kenya with a solar heating water system in order to provide hot water for the Maternity and Paediatrics wards that host 79 beds. Capacity of the system is 600 litres of hot water in Maternity and 600 litres in Paediatrics. This project, led by Ms. Beatriz Meijide, Camusat Finance Director for the Africa region, who has been personally volunteering in the hospital for 15 years, was part of Camusat’s employees ‘associative commitment incentive program, named “I volunteer with Camusat” and launched in December 2015. “As the His Lordship the Bishop of Embu said on the occasion of H. E. Margaret Kenyatta’s visit to the hospital: “[…] Regardless of who you are, regardless of what state… we can make a difference by our contribution. What you give, that self-giving to serve others… Any effort you make to change the lives of others, bears fruit”. That has been my motivation, and indeed I have witnessed an amazing turnaround of this hospital, that has helped to save so many lives thanks to the effort of the great staff and management they have, and the contributions of so many friends and supporters. Now Camusat has marked a new milestone with the provision of hot water, something so essential for the maternal & child healthcare. For this I am deeply grateful and proud to work in a company that really walks the talk, and has made this long term awaited project come true for the benefit of the patients” said Ms. Beatriz Meijide. “The « I volunteer with Camusat » program reiterates our commitment in investing in the development and recognition within our host communities, a pillar of our CSR policy. Through this support, we also hope to encourage our people in their personal and associative efforts to promote education, healthcare access and environment protection. The project led by Beatriz drew our attention since it aims at offering better access to healthcare thanks to a green energy solution, one of our areas of expertise” stated Elodie Perrigot, HR-CSR & EHS Chief Officer of Camusat Group. Thanks to this solar heating water system, Consolata hospital, which has been awarded the Johnson and Johnson’s Best Maternity in Eastern, can now offer even better antenatal and postnatal care; an improvement that the whole community will benefit from. “The system is now working and is able to heat five showers in Maternity and four in Paediatrics. Since its commissioning, the system has been serving an average of 60 babies in paediatrics ward and 80 mothers in maternity daily. We highly appreciate the contribution of Camusat towards this noble cause which will go a long way in improving the overall hygiene” said Dominic Muriuki Mugo, Consolata Hospital Managing Director. Please visit our website to see pictures of this action: http://www.camusat.com/en/news/ Located on Kenya's Eastern Province, under the Diocese of Embu since 1970, it comprises of 167 beds, outpatient facilities and 15 dispensaries in remote areas. Also a teaching hospital, it hosts the FIDENZA School of Nursing. The A&E casualty department is open 24 hours a day providing comprehensive emergency services as well as receiving patients referred from primary health care facilities and clinics in the environs. CAMUSAT is one of the market leaders in the implementation of telecom infrastructures and one of the most experienced network developers with a history which starts back in the late 40’s Our committed mission is to help the telecom actors to efficiently build and optimize their networks development by integrating the traditional telecom technologies with the most innovative know-how from adjoining fields. Our R&D Department develops innovative infrastructure solutions. Integration of quality-proofed technologies and services, as well as proficient project management skills and flexibility have played a crucial part in our success today. Diversity is another key success factor. Motivating incentive plans, strong team spirit, constant professional training, international mobility and efficiency are our strengths in achieving our client’s high standard requirements. Camusat’s headquarters are located in Paris, France. Nearly half of Camusat's 2000 employees work on the African continent. The company operates on the five continents with over 40 subsidiaries.
News Article | October 29, 2016
- El taller patrocinado por Ascensia Diabetes Care en ISPAD 2016 destaca la importancia de la tecnología digital para apoyar el autotratamiento de la diabetes en pediatría y adolescentes Hoy, en un taller organizado por Ascensia Diabetes Care, un reconocido panel de expertos destacó la importancia de la tecnología digital para apoyar el auto-tratamiento de la diabetes en pediatría y adolescentes. El taller se llevó a cabo en el 42 encuentro anual de la International Society for Pediatric and Adolescent Diabetes (ISPAD), que está teniendo lugar del 26 al 29 de octubre de 2016 en Valencia (España). El evento, titulado "Smart Diabetes Self-Management in Paediatrics and Adolescents in the Digital Age", examina el uso de las tecnologías digitales y salud móvil para apoyar el auto-tratamiento de la diabetes en los niños. Concluyó que la tecnología móvil tiene un importante potencial para apoyar el auto-tratamiento en esta importante población. Sin embargo, la calidad y precisión de las herramientas de salud móvil en la diabetes se destacaron como consideraciones importantes. Hay muchas herramientas que están actualmente disponibles en el mercado y es importante identificar las aplicaciones de salud móvil que son seguras y fiables para el tratamiento de niños y adolescentes con diabetes. Presidiendo el taller, el doctor Henk Veeze, director médico internacional senior en Diabeter and Immediate Past Treasurer de ISPAD, explicó: "Sabemos que la diabetes puede ser difícil de tratar a cualquier edad, pero es particularmente complicado para los jóvenes, ya que están creciendo. Estos retos pueden afectar significativamente a su salud y conducir a complicaciones y pobres resultados. Las herramientas digitales y dispositivos móviles tienen el potencial de mejorar significativamente la gestión de la diabetes para los jóvenes y sus padres, ayudando a seguir la condición, ofreciendo información accionable y apoyando el auto-tratamiento". El taller incluyó presentaciones de Javier Ferrero, director de información de la Agencia Andaluza de Calidad Asistencial en España, y el doctor David Montes, endocrinólogo pediátrico sénior del Hospital de Fuenlabrada en España. "El objetivo de las nuevas tecnologías digitales en la diabetes es mejorar el control metabólico de los niños, y las aplicaciones desempeñan un papel importante en este objetivo apoyando a los niños y a sus padres en la toma de decisiones. Es por tanto esencial que mejoremos las herramientas digitales que están disponibles para ellos. Los pacientes infantiles y sus padres necesitan herramientas y aplicaciones móviles efectivas y precisas que apoyen el auto-tratamiento", explicó el doctor David Montes. En relación con los estándares y regulaciones para las tecnologías de aplicaciones móviles en la diabetes, Javier Ferrero añadió: "Las aplicaciones de diabetes tienen el enorme potencial de cambiar drásticamente las conductas y hábitos de los pacientes, empoderándoles a gestionar mejor su enfermedad. Sin embargo, muchas aplicaciones de diabetes no cumplen los estándares de mejores prácticas de las tecnologías digitales modernas. Por ejemplo, las aplicaciones no seguras suponen un riesgo muy importante en las manos de los pacientes más jóvenes que no tienen la capacidad de diferenciar entre aplicaciones fiables y valiosas. La regulación y la vigilancia del mercado deben mejorarse de manera urgente para garantizar la seguridad del paciente". ISPAD quiere promover la ciencia clínica y básica, la investigación, la educación y el derecho en la diabetes en niños y adolescentes, y es la única sociedad internacional centrada específicamente en todos los tipos de diabetes infantil. El tema del encuentro 2016 se basa en la educación y en las nuevas tecnologías para mejorar el cuidado de la diabetes. Michael Kloss, consejero delegado de Ascensia Diabetes Care, dijo: "Estamos muy contentos de asistir a la Conferencia anual de ISPAD 2016 y ofrecer este taller educativo sobre la importancia de las tecnologías digitales para apoyar la atención de la diabetes infantil. Como socio de confianza en la comunidad de la diabetes, estamos dedicados a apoyar la educación sobre este tema crucial". ISPAD 2016 también coincide con el lanzamiento de los sistemas de monitorización de la glucosa en sangre CONTOUR®NEXT ONE y CONTOUR®PLUS ONE por Ascensia Diabetes Care en Europa. Cada sistema incluye un medidor inalámbrico y fácil de utilizar que se vincula sin fisuras a un dispositivo móvil inteligente a través de la conectividad Bluetooth®. Los datos que se capturan sin esfuerzo por la aplicación CONTOUR® DIABETES pueden evaluarse para tendencias en los niveles de glucosa sanguínea y ofrecen información accionable. Michael Kloss añadió: "La aplicación CONTOUR DIABETES habilita patrones y tendencias en los niveles de glucosa en sangre en niños con diabetes que deben mostrarse a los mismos niños, a sus padres y a sus médicos. Esto puede ayudar a apoyar el auto-tratamiento por toda la familia y el equipo sanitario".