One Night of Partial Sleep Deprivation Affects Biomarkers of Cardiac Damage, but Not Cardiovascular and Lipid Profiles, in Young Athletes

Sleep loss is among the most common yet frequently overlooked problems. This disruptive influence is associated with an adverse lipid profile (LP) and consequently results in an increased risk of cardiovascular disease. Furthermore, it has been well established that athletes are increasingly confronted with sleep problems. The aim of this study was to explore the effect of one night of partial sleep deprivation (PSD) on the cardiovascular profile and LP in young, trained athletes. Ten male Taekwondo athletes were randomized for three sleep conditions in a counterbalanced order: (i) following a baseline sleep night (BN), (ii) following PSD at the beginning of the night (PSDBN), and (iii) following PSD at the end of the night (PSDEN). Basal cardiovascular physiological measures were recorded, and blood samples were taken in the fasted state following each sleep session (i.e., in the morning at 07:00 h). The results showed that myoglobin and creatine phosphokinase increased significantly after PSDEN but not after PSDBN. By contrast, no alteration was observed in the LP and physiological parameters following the two types of PSD. In conclusion, these results show that PSDEN increases cardiac damage biomarkers significantly, even though they do not reach clinical significance. Thus, one night of PSD does not affect the physiological responses and biomarkers of LP in Taekwondo athletes

Uptake of intermittent preventive treatment and pregnancy outcomes: health facilities and community surveys in Chókwè district, southern Mozambique

Abstract Background Malaria in pregnancy leads to serious adverse effects on the mother and the child and accounts for 75,000–200,000 infant deaths every year. Currently, the World Health Organization recommends intermittent preventive treatment of malaria in pregnancy (IPTp) with sulfadoxine–pyrimethamine (SP) at each scheduled antenatal care (ANC) visit. This study aimed to assess IPTp-SP coverage in mothers delivering in health facilities and at the community. In addition, factors associated with low IPTp-SP uptake and malaria adverse outcomes in pregnancy were investigated. Methods A community and a health facility-based surveys were conducted in mothers delivering in Chókwè district, southern Mozambique. Social-demographic data, malaria prevention practices and obstetric history were recorded through self-report and antenatal records. For women delivering at health facilities, a clinical examination of mother and child was performed, and malaria infection at delivery was determined by rapid diagnostic test, microscopy, quantitative PCR and placental histology. Results Of 1141 participants, 46.6, 30.2, 13.5 and 9.6% reported taking ≥ 3, two, one and none SP doses, respectively. Low IPTp uptake (< 3 doses) was associated with non-institutional deliveries (AOR = 2.9, P < 0.001), first ANC visit after week 28 (AOR = 5.4, P < 0.001), low awareness of IPTp-SP (AOR = 1.6, P < 0.002) and having no or only primary education (AOR = 1.3, P = 0.041). The overall prevalence of maternal malaria (peripheral and/or placental) was 16.8% and was higher among women from rural areas compared to those from urban areas (AOR = 1.9, P < 0.001). Younger age (< 20 years; AOR = 1.6, P = 0.042) and living in rural areas (AOR = 1.9, P < 0.001) were predictors of maternal malaria at delivery. Being primigravidae (AOR = 2.2, P = 0.023) and preterm delivery (AOR = 2.6, P < 0.001) predicted low birth weight while younger age was also associated with premature delivery (AOR = 1.4, P = 0.031). Conclusion The coverage for two and ≥ 3 doses of IPTp-SP is moderately higher than estimates from routine health facility records in Gaza province in 2015. However, this is still far below the national target of 80% for ≥ 3 doses. Ongoing campaigns aiming to increase the use of malaria prevention strategies during pregnancy should particularly target rural populations, increasing IPTp-SP knowledge, stimulate early visits to ANC, improve access to health services and the quality of the service provided

Molecular surveillance of Plasmodium falciparum drug-resistance markers in Vietnam using multiplex amplicon sequencing (2000–2016)

Abstract Emergence and spread of Plasmodium falciparum resistance to artemisinin-based combination therapies (ACT) is a major challenge for Greater Mekong Subregion countries in their goal to eliminate malaria by 2030. Tools to efficiently monitor drug resistance beyond resource-demanding therapeutic efficacy studies are necessary. A custom multiplex amplicon sequencing assay based on Illumina technology was designed to target the marker of partial resistance to artemisinin (K13), five candidate modulators of artemisinin resistance, the marker of resistance to chloroquine (crt), and four neutral microsatellite loci. The assay was used to genotype 635 P. falciparum-positive blood samples collected across seven provinces of Vietnam and one of Cambodia between 2000 and 2016. Markers of resistance to artemisinin partner-drugs piperaquine (copy number of plasmepsin-2) and mefloquine (copy number of multidrug-resistance 1) were determined by qPCR. Parasite population structure was further assessed using a 101-SNP barcode. Validated mutations of artemisinin partial resistance in K13 were found in 48.1% of samples, first detection was in 2000, and by 2015 prevalence overcame > 50% in Central Highlands and Binh Phuoc province. K13-C580Y variant became predominant country-wide, quickly replacing an outbreak of K13-I543T in Central Highlands. Mutations in candidate artemisinin resistance modulator genes paralleled the trends of K13 mutants, whereas resistance to piperaquine and mefloquine remained low (≈ 10%) by 2015–2016. Genomic tools applied to malaria surveillance generate comprehensive information on dynamics of drug resistance and population structure and reflect drug efficacy profiles from in vivo studies

MOESM1 of Uptake of intermittent preventive treatment and pregnancy outcomes: health facilities and community surveys in Chókwè district, southern Mozambique

Additional file 1: Table S1. Characteristics of the study population by delivering place and associations with non-institutional delivery

Prevalence, Specificity and Determinants of Lipid-Interacting PDZ Domains from an In-Cell Screen and <em>In Vitro</em> Binding Experiments

<div><h3>Background</h3><p>PDZ domains are highly abundant protein-protein interaction modules involved in the wiring of protein networks. Emerging evidence indicates that some PDZ domains also interact with phosphoinositides (PtdInsPs), important regulators of cell polarization and signaling. Yet our knowledge on the prevalence, specificity, affinity, and molecular determinants of PDZ-PtdInsPs interactions and on their impact on PDZ-protein interactions is very limited.</p> <h3>Methodology/Principal Findings</h3><p>We screened the human proteome for PtdInsPs interacting PDZ domains by a combination of <em>in vivo</em> cell-localization studies and <em>in vitro</em> dot blot and Surface Plasmon Resonance (SPR) experiments using synthetic lipids and recombinant proteins. We found that PtdInsPs interactions contribute to the cellular distribution of some PDZ domains, intriguingly also in nuclear organelles, and that a significant subgroup of PDZ domains interacts with PtdInsPs with affinities in the low-to-mid micromolar range. <em>In vitro</em> specificity for the head group is low, but with a trend of higher affinities for more phosphorylated PtdInsPs species. Other membrane lipids can assist PtdInsPs-interactions. PtdInsPs-interacting PDZ domains have generally high pI values and contain characteristic clusters of basic residues, hallmarks that may be used to predict additional PtdInsPs interacting PDZ domains. In tripartite binding experiments we established that peptide binding can either compete or cooperate with PtdInsPs binding depending on the combination of ligands.</p> <h3>Conclusions/Significance</h3><p>Our screen substantially expands the set of PtdInsPs interacting PDZ domains, and shows that a full understanding of the biology of PDZ proteins will require a comprehensive insight into the intricate relationships between PDZ domains and their peptide and lipid ligands.</p> </div

Distinct distributions of human eYFP-S1 PDZ1-PDZX in MCF-7 cells as determined by wide-field and/or confocal microscopy.

<p>Distinct distributions of human eYFP-S1 PDZ1-PDZX in MCF-7 cells as determined by wide-field and/or confocal microscopy.</p

Lipid binding profiles of PDZ domains and differential effects of peptide ligands on PtdInsPs binding.

<p><b>A</b>. Apparent PDZ-PtdInsPs affinities as determined by SPR equilibrium binding experiments between his-tagged PDZ domains and 5% PtdInsPs (DOPC liposomes). The color code is indicated to the right. Dark blue color indicates that values were not determinable, for apparent K_D values see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0054581#pone.0054581.s006" target="_blank">Table S2</a>. <b>B</b>. Schematics of the pre-spotted lipid blot membrane together with representative lipid blots of DFNB31_1, MAGI3_3 and MAGI1_6. <b>C</b>. Overview of the lipid blot analysis. Detected spots are represented by black filled circles for intense signals or grey for weaker signals, as compared within each membrane. No binding was detected for TGA, DGA, PC, cholesterol or sphingomyelin. <b>D.</b> Double reference subtracted sensorgrams of DFNB31_1, MAGI1_6 and MAGI3_3 injected over 10% PS containing DOPC liposomes. The highest protein concentration used is indicated for each protein. <b>E.</b> Equilibrium peptide binding titrations of DFNB31_1/Y167W and the C-terminal peptide of usher (FTDTHL, black circles and black line), and CASK/I517W and the C-terminal peptide of syndecan2 (TKEFYA, blue squares and blue line) as followed by changes in intrinsic tryptophan fluorescence. The protein concentrations were kept constant at 3 µM. <b>F</b>. Equilibrium binding isotherms of DNB31_1 (left panel) and CASK (right panel) to 5% PtdIns(4,5)P2 (DOPC liposomes) as determined by SPR experiments in absence (black lines) and presence (red lines) of 1 mM of respective peptide ligands. Note the apparent decrease and increase in affinity in presence of peptide for DFNB31_1 and CASK, respectively.</p

Subcellular distribution of PDZ domains and effects of PtdInsP-modifying treatments compared to <i>in vitro</i> PtdIns(4,5)P2-binding.

<p><b>A.</b> Double reference subtracted sensorgrams of recombinant his-tagged PDZ domains injected, in a range of concentrations with the highest concentrations used indicated, over 5% PtdIns(4,5)P2 containing DOPC liposomes. <b>B–O.</b> Confocal or wide-field micrographs of MCF-7 cells transiently over-expressing selected eYFP-S1PDZ1-tagged PDZ domains (with the exception of SLC9A3R2_1* where S1PDZ1 was omitted). Sensorgrams are provided to the left of the micrographs for each PDZ domain under study. <b>B</b>. Co-expression of eYFP-S1PDZ1-MPP7 with PtdIns(4)P-5-kinase (PIPK; shown in the insets) to probe the effect of increased plasma membrane PtdIns(4,5)P2 levels on the distribution of the fluorescent protein. Note the absence of membrane enrichment of the fluorescence upon kinase over-expression. <b>C</b>. Serum stimulation of eYFP-S1PDZ1-MPP7 expressing cells to investigate the effect of increased plasma membrane PtdIns(3,4,5)P3 levels. Note the absence of membrane enrichment of the fluorescence upon serum stimulation. For control experiments of B and C, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0054581#pone.0054581.s003" target="_blank">Fig. S3A</a>–B. <b>D–G</b> Ionomycin treatment (D, E), inhibition of PtdIns(4)-kinases by PAO (F) and rapamycin-induced membrane recruitment of PtdInsPs 5′ phosphatase to investigate how reducing PtdIns(4,5)P2 levels affect the membrane enrichment of eYFP-S1PDZ1-MPDZ_7 (D) and eYFP-S1PDZ1-CASK (E-G). <b>H–I.</b> Co-expression with mCherry-PTS1 identified the bright cytosolic spots enriched in eYFP-S1PDZ1-MAGI3_3 (H) and eYFP-S1PDZ1-IL16_1 (I) as peroxisomes. Co-expression with mCherry-B23 established that the subnuclear organelles enriched in eYPF-S1PDZ1-SCRIB4 (J) and eYFP-SLC9A3R2_1* (M) corresponded to nucleoli. Co-expression with a PtdIns(4,5)P2-4-phosphatase (PIP2 4 ptase, shown in the insets) or PCLΔNES, did not affect the nucleoli enrichment of eYFP-S1PDZ1-SCRIB4 (K, L) but induced a shift of eYFP-SLC9A3R2_1 (N, O) towards the nucleoplasm and the cytosol.</p