Multiple pathways for SARS-CoV-2 resistance to nirmatrelvir.

Nirmatrelvir, an oral antiviral targeting the 3CL protease of SARS-CoV-2, has been demonstrated to be clinically useful against COVID-19 (refs. 1,2). However, because SARS-CoV-2 has evolved to become resistant to other therapeutic modalities3-9, there is a concern that the same could occur for nirmatrelvir. Here we examined this possibility by in vitro passaging of SARS-CoV-2 in nirmatrelvir using two independent approaches, including one on a large scale. Indeed, highly resistant viruses emerged from both and their sequences showed a multitude of 3CL protease mutations. In the experiment peformed with many replicates, 53 independent viral lineages were selected with mutations observed at 23 different residues of the enzyme. Nevertheless, several common mutational pathways to nirmatrelvir resistance were preferred, with a majority of the viruses descending from T21I, P252L or T304I as precursor mutations. Construction and analysis of 13 recombinant SARS-CoV-2 clones showed that these mutations mediated only low-level resistance, whereas greater resistance required accumulation of additional mutations. E166V mutation conferred the strongest resistance (around 100-fold), but this mutation resulted in a loss of viral replicative fitness that was restored by compensatory changes such as L50F and T21I. Our findings indicate that SARS-CoV-2 resistance to nirmatrelvir does readily arise via multiple pathways in vitro, and the specific mutations observed herein form a strong foundation from which to study the mechanism of resistance in detail and to inform the design of next-generation protease inhibitors.

Molecular mechanisms of SARS-CoV-2 resistance to nirmatrelvir.

Nirmatrelvir is a specific antiviral drug that targets the main protease (Mpro) of SARS-CoV-2 and has been approved to treat COVID-191,2. As an RNA virus characterized by high mutation rates, whether SARS-CoV-2 will develop resistance to nirmatrelvir is a question of concern. Our previous studies have shown that several mutational pathways confer resistance to nirmatrelvir, but some result in a loss of viral replicative fitness, which is then compensated for by additional alterations3. The molecular mechanisms for this observed resistance are unknown. Here we combined biochemical and structural methods to demonstrate that alterations at the substrate-binding pocket of Mpro can allow SARS-CoV-2 to develop resistance to nirmatrelvir in two distinct ways. Comprehensive studies of the structures of 14 Mpro mutants in complex with drugs or substrate revealed that alterations at the S1 and S4 subsites substantially decreased the level of inhibitor binding, whereas alterations at the S2 and S4' subsites unexpectedly increased protease activity. Both mechanisms contributed to nirmatrelvir resistance, with the latter compensating for the loss in enzymatic activity of the former, which in turn accounted for the restoration of viral replicative fitness, as observed previously3. Such a profile was also observed for ensitrelvir, another clinically relevant Mpro inhibitor. These results shed light on the mechanisms by which SARS-CoV-2 evolves to develop resistance to the current generation of protease inhibitors and provide the basis for the design of next-generation Mpro inhibitors.

High-Density Proximity Mapping Reveals the Subcellular Organization of mRNA-Associated Granules and Bodies.

mRNA processing, transport, translation, and ultimately degradation involve a series of dedicated protein complexes that often assemble into large membraneless structures such as stress granules (SGs) and processing bodies (PBs). Here, systematic in vivo proximity-dependent biotinylation (BioID) analysis of 119 human proteins associated with different aspects of mRNA biology uncovers 7424 unique proximity interactions with 1,792 proteins. Classical bait-prey analysis reveals connections of hundreds of proteins to distinct mRNA-associated processes or complexes, including the splicing and transcriptional elongation machineries (protein phosphatase 4) and the CCR4-NOT deadenylase complex (CEP85, RNF219, and KIAA0355). Analysis of correlated patterns between endogenous preys uncovers the spatial organization of RNA regulatory structures and enables the definition of 144 core components of SGs and PBs. We report preexisting contacts between most core SG proteins under normal growth conditions and demonstrate that several core SG proteins (UBAP2L, CSDE1, and PRRC2C) are critical for the formation of microscopically visible SGs.

Quantifying the clusterness and trajectoriness of single-cell RNA-seq data.

Among existing computational algorithms for single-cell RNA-seq analysis, clustering and trajectory inference are two major types of analysis that are routinely applied. For a given dataset, clustering and trajectory inference can generate vastly different visualizations that lead to very different interpretations of the data. To address this issue, we propose multiple scores to quantify the "clusterness" and "trajectoriness" of single-cell RNA-seq data, in other words, whether the data looks like a collection of distinct clusters or a continuum of progression trajectory. The scores we introduce are based on pairwise distance distribution, persistent homology, vector magnitude, Ripley's K, and degrees of connectivity. Using simulated datasets, we demonstrate that the proposed scores are able to effectively differentiate between cluster-like data and trajectory-like data. Using real single-cell RNA-seq datasets, we demonstrate the scores can serve as indicators of whether clustering analysis or trajectory inference is a more appropriate choice for biological interpretation of the data.

Stromal STAT5-Mediated Trophic Activity Regulates Hematopoietic Niche Factors.

Signal transducer and activator of transcription 5 (STAT5a and STAT5b) are intrinsically critical for normal hematopoiesis but are also expressed in stromal cells. Here, STAT5ab knockout (KO) was generated with a variety of bone marrow hematopoietic and stromal Cre transgenic mouse strains. Vav1-Cre/+STAT5abfl/fl, the positive control for loss of multipotent hematopoietic function, surprisingly dysregulated niche factor mRNA expression, and deleted STAT5ab in CD45neg cells. Single-cell transcriptome analysis of bone marrow from Vav1-Cre/+ wild-type or Vav1-Cre/+STAT5abfl/fl mice showed hematopoietic stem cell (HSC) myeloid commitment priming. Nes+ cells were detected in both CD45neg and CD45+ clusters and deletion of STAT5ab with Nes-Cre caused hematopoietic repopulating defects. To follow up on these promiscuous Cre promoter deletions in CD45neg and CD45+ bone marrow cell populations, more stroma-specific Cre strains were generated and demonstrated a reduction in multipotent hematopoietic progenitors. Functional support for niche-supporting activity was assessed using STAT5-deficient mesenchymal stem cells (MSCs). With Lepr-Cre/+STAT5abfl/fl, niche factor mRNAs were downregulated with validation of reduced IGF-1 and CXCL12 proteins. Furthermore, advanced computational analyses revealed a key role for STAT5ab/Cish balance with Cish strongly co-expressed in MSCs and HSCs primed for differentiation. Therefore, STAT5ab-associated gene regulation supports the bone marrow microenvironment.

Analysis of retinal and choroidal microvascular changes using optical coherence tomography and optical coherence tomography angiography in patients with acute leukemia.

PURPOSE: Although leukemic retinopathy accounts for 80% of ocular complications in acute leukemia, its pathogenesis remains unclear. To evaluate changes in retinal and choroicapillaris and structural parameters in patients with acute leukemia, we analyzed the correlation between vascular perfusion metrics and laboratory parameters and assessed the changes after hematopoietic stem cell transplantation (HSCT). METHODS: Herein, 104 eyes of 52 patients aged 18 and above with acute leukemia were enrolled. 80 eyes of 40 healthy patients were recruited as control participants. All participants underwent optical coherence tomography (OCT) and OCT angiography (OCTA) at baseline. RESULTS: Patients with acute leukemia had a significantly thicker ganglion cell-inner plexiform layer (GCIPL) and lower circularity index than the control participants. Post-HSCT perfusion metrics did not differ significantly, but parafoveal thickness decreased significantly. During the active phase of acute leukemia, lower platelet levels were associated with significant GCIPL thickening and increased foveal avascular zone and perimeter. D-dimer levels positively correlated with GCIPL thickness. CONCLUSION: Patients with acute leukemia had subclinical retinal microvascular deficits on OCTA and GCIPL thickening on OCT, possibly associated with bone marrow function. GCIPL thickness may indicate acute ischemia in such patients. Further studies must elucidate their clinical and prognostic significance.

Factors associated with postpartum depression symptoms among postpartum women in five countries during the COVID-19 pandemic: an online cross-sectional study.

BACKGROUND: This study aimed to examine factors associated with postpartum depression (PPD) symptoms during the COVID-19 pandemic among postpartum women in five countries, a subject that has not been investigated thus far. METHODS: A multi-country, cross-sectional, online survey was conducted with a convenience sample of 3,523 postpartum women in Brazil, South Korea, Taiwan, Thailand, and the United Kingdom, from July to November 2021. Sociodemographic and obstetric data, food insecurity, COVID-19 positive status, COVID-19 vaccination, infant feeding, breastfeeding belief score, and social support were investigated. PPD and social support were measured using the Edinburgh Postnatal Depression Scale and Maternal Social Support Scale, respectively. Descriptive statistics, chi-squared tests, and t-tests were used to identify associations with PPD symptoms. A binary logistic regression model was used to identify explanatory factors associated with PPD and adjusted odds ratios (OR) and 95% confidence intervals (CIs) were calculated. RESULTS: Women in Taiwan (AOR = 0.5; 95%CI 0.34, 0.73) and Thailand (AOR = 0.68; 95%CI 0.46, 0.99) had a lower risk of PPD symptoms than those in Brazil. In addition, women with planned pregnancies had a lower risk of PPD (AOR = 0.74; 95%CI 0.60, 0.91). Younger women (AOR = 1.62; 95%CI 1.05, 2.51), health problems during pregnancy, delivery, or postpartum (AOR = 1.71; 95%CI 1.42, 2.06), and no change or worse food insecurity during COVID-19 (AOR = 1.66; 95%CI 1.21, 1.27 for no change and AOR = 1.68; 95%CI 1.27, 1.23, respectively) presented a higher likelihood of having PPD. Feeding babies with expressed human milk (AOR = 1.25; 95%CI 1.03, 1.50) and/or complementary food (AOR = 1.51; 95%CI 1.17, 1.94) were associated with PPD symptoms. Women who received low (AOR = 7.74; 95%CI 5.43, 11.03) or medium support (AOR = 3.25; 95%CI 2.71, 3.88) had higher likelihoods of PPD. CONCLUSION: PPD symptoms during the pandemic were high in young women, particularly Brazilian women, with health problems in the puerperal pregnancy cycle who fed their babies expressed breast milk and/or complementary food. Low social support also impacted PPD symptoms. This study highlights the need for the professional screening for PPD and provision of virtual or personal support.

Prevalence of adolescent-reported food insecurity and the determinants including coping strategies living in urban slum communities of Bangladesh during the era of COVID-19: a cross-sectional study.

BACKGROUND: As food insecurity (FI) continues to rise worldwide especially in developing countries like Bangladesh, adolescent experience of FI have received minimal attention globally. This study aimed to identify the prevalence of adolescent-reported FI and its association with individual and socio-environmental factors as well as coping strategies amongst a sample of adolescents living in urban slum areas of Bangladesh in the times of the Coronavirus 19 (COVID-19) pandemic. METHODS: A descriptive cross sectional study was conducted amongst 326 adolescents (12-18 years) living in the urban slums of Narayanganj, Dhaka from April to May, 2022. Adolescent-reported FI was assessed using a structured questionnaire adopted from Household Food Insecurity Access Scale (HFIAS). Descriptive statistics, Chi-square tests and ordinal logistic regression were used to draw inference. RESULTS: Prevalence of adolescent-reported FI was high (46.6% moderate and 29.8% severe). The likelihood of experiencing moderate or severe FI versus no/mild FI were 1.7 times (95% Confidence Interval (CI) [1.1, 2.5]) higher in younger adolescents and 5 times (95% CI [2.3, 12.7]) higher in unemployed youth. Socio-environmental factors determining the economic status of a household such as higher number of family members, only one earning family member, unemployed father, no household assets, food aid received by the family during pandemic and positive COVID-19 infection in family were associated with moderate and severe FI. Coping strategies such as a higher number of food seeking strategies (Adjusted Odds Ratio (AOR) 3.4, 95% CI [1.9, 5.9]), substance use (AOR 6.2, 95% CI [1.2, 31.7]) and stopping school (AOR 3.3, 95% CI [1.9, 5.7]) increased odds for moderate and severe FI. Stratified by drop-out of school status, an association between food seeking strategies and FI remained significant among those school-going, while there was no association among those dropping out of school. CONCLUSION: This study showed that adolescents living in urban slum communities in Bangladesh are at very high risk of FI and resort to harmful coping strategies. Our study highlights the importance of further research in adolescent reported FI and coping strategies in low to middle income countries (LMICs) and create appropriate interventions to lower FI among this group and improve their state of health and wellbeing.

Associations between breastfeeding intention, breastfeeding practices and post-natal depression during the COVID-19 pandemic: A multi-country cross-sectional study.

Associations between breastfeeding intention, duration and post-natal depression (PND) have been shown in pre-COVID-19 studies. However, studies during COVID-19 have not examined the associations between breastfeeding intention, breastfeeding practices, and PND in an international sample of post-natal women, taking into consideration COVID-19 related factors. This is the first study to address this gap as both PND and breastfeeding may be affected by COVID-19, and have important long-term effects on women's and infant's health. A cross-sectional internet-based survey was conducted with 3253 post-natal women from five countries: Brazil, South Korea, Taiwan, Thailand, and the United Kingdom from July to November 2021. The results showed that women who intended to breastfeed during pregnancy had lower odds of having PND than women who did not intend to. Women who had no breastfeeding intention but actually breastfed had greater odds (AOR 1.75) of having PND than women who intended to breastfeed and actually breastfed. While there was no statistical significance in expressed breast milk feeding in multivariable logistic regression models, women who had shorter duration of breastfeeding directly on breast than they planned had greater odds (AOR 1.58) of having PND than those who breastfed longer than they planned even after adjusting for covariates including COVID-19-related variables. These findings suggested the importance of working with women on their breastfeeding intention. Tailored support is required to ensure women's breastfeeding needs are met and at the same time care for maternal mental health during and beyond the pandemic.

Inhibitors of Coronavirus 3CL Proteases Protect Cells from Protease-Mediated Cytotoxicity.

We describe a mammalian cell-based assay to identify coronavirus 3CL protease (3CLpro) inhibitors. This assay is based on rescuing protease-mediated cytotoxicity and does not require live virus. By enabling the facile testing of compounds across a range of 15 distantly related coronavirus 3CLpro enzymes, we identified compounds with broad 3CLpro-inhibitory activity. We also adapted the assay for use in compound screening and in doing so uncovered additional severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) 3CLpro inhibitors. We observed strong concordance between data emerging from this assay and those obtained from live-virus testing. The reported approach democratizes the testing of 3CLpro inhibitors by developing a simplified method for identifying coronavirus 3CLpro inhibitors that can be used by the majority of laboratories, rather than the few with extensive biosafety infrastructure. We identified two lead compounds, GC376 and compound 4, with broad activity against all 3CL proteases tested, including 3CLpro enzymes from understudied zoonotic coronaviruses. IMPORTANCE Multiple coronavirus pandemics have occurred over the last 2 decades. This has highlighted a need to be proactive in the development of therapeutics that can be readily deployed in the case of future coronavirus pandemics. We developed and validated a simplified cell-based assay for the identification of chemical inhibitors of 3CL proteases encoded by a wide range of coronaviruses. This assay is reporter free, does not require specialized biocontainment, and is optimized for performance in high-throughput screening. By testing reported 3CL protease inhibitors against a large collection of 3CL proteases with variable sequence similarity, we identified compounds with broad activity against 3CL proteases and uncovered structural insights into features that contribute to their broad activity. Furthermore, we demonstrated that this assay is suitable for identifying chemical inhibitors of proteases from families other than 3CL proteases.

JSOM: Jointly-evolving self-organizing maps for alignment of biological datasets and identification of related clusters.

With the rapid advances of various single-cell technologies, an increasing number of single-cell datasets are being generated, and the computational tools for aligning the datasets which make subsequent integration or meta-analysis possible have become critical. Typically, single-cell datasets from different technologies cannot be directly combined or concatenated, due to the innate difference in the data, such as the number of measured parameters and the distributions. Even datasets generated by the same technology are often affected by the batch effect. A computational approach for aligning different datasets and hence identifying related clusters will be useful for data integration and interpretation in large scale single-cell experiments. Our proposed algorithm called JSOM, a variation of the Self-organizing map, aligns two related datasets that contain similar clusters, by constructing two maps-low-dimensional discretized representation of datasets-that jointly evolve according to both datasets. Here we applied the JSOM algorithm to flow cytometry, mass cytometry, and single-cell RNA sequencing datasets. The resulting JSOM maps not only align the related clusters in the two datasets but also preserve the topology of the datasets so that the maps could be used for further analysis, such as clustering.

Discovery of a simplified deguelin analog as an HSP90 C-terminal inhibitor for HER2-positive breast cancer.

A series of O-substituted analogs of the C-ring-truncated scaffold of deguelin designed as heat shock protein 90 (HSP90) C-terminal inhibitors were investigated as novel antitumor agents against human epidermal growth factor receptor 2 (HER2)-positive breast cancer. Among the synthesized compounds, compound 37 displayed significant inhibition in both trastuzumab-sensitive and trastuzumab-resistant breast cancer cells with little cytotoxicity to normal cells. Mechanistic studies of compound 37 carried out by HSP90α C-terminal inhibitor screening, the induction of the heat shock response and downregulation of HSP90 client proteins indicated that the antitumor activity of 37 in breast cancer cells could be attributed to the destabilization and inactivation of HSP90 client proteins by the binding of 37 to the C-terminal domain of HSP90. A molecular docking study of compound 37 with a HSP90 homology model indicated that its S-isomer fit well in the ATP binding site of the C-terminal domain, forming key interactions.

Prevalence and regional variations of coexistence of child stunting and maternal overweight or obesity in Myanmar.

OBJECTIVE: The current study aimed to investigate double burden of malnutrition within households at the national and subnational levels and to identify its association with sociodemographic factors in Myanmar. DESIGN: All the variables were extracted from children's file of the Myanmar Demographic and Health Survey 2015-2016. Children under five were identified as stunted based on a height-for-age < -2 sd below the WHO reference median. Maternal overweight/obesity was defined as a BMI ≥ 23 kg/m2. A stunted child with an overweight/obese mother (SCOM) was classified as a double-burden household. SETTING: A national household survey in Myanmar. PARTICIPANTS: Children under five and their mothers (n 3954 pairs). RESULTS: Mean ages of children and mothers were 29 (se 0·14) months and 30·9 (se 0·32) years, respectively. National prevalence of childhood stunting and maternal overweight/obesity was 28·0 % and 39·4 %, respectively, and the prevalence of SCOM was 9·1 %. Significant regional differences were found in SCOM, ranging from 3·6 % in Naypyitaw to 12 % in Kachin and Mon and 14·6 % in Kayah. In the multinomial logistic regression analysis, relative to neither a stunted child nor an overweight/obese mother, child's age, maternal age, maternal experience of a teen birth, short mothers, mothers with primary education and in middle or rich wealth tertiles, and some regions (Kachin, Kayah, Shan, Sagaing, Taninthayi, Ayeyarwaddy, Mon and Yangon) were associated with greater odds of SCOM. CONCLUSIONS: The current study showed a relatively high national prevalence of SCOM and significant regional variations. Overarching policies and programmes with culturally sensitive strategies need to be formulated and implemented.

Double burden of malnutrition and its association with infant and young child feeding practices among children under-five in Thailand.

OBJECTIVE: This study examined the prevalence of stunting-overweight and socio-demographic determinants among children under-five years of age, as well as associations with infant and young child feeding (IYCF) among children aged 6-23 months. DESIGN: Secondary data analysis based on the Thailand Multiple Indicator Cluster Survey 2015-2016. SETTING: Cross-national study. PARTICIPANTS: Nationally representative sample of children under-five years of age (n 12 313). RESULTS: The prevalence of wasting, stunting, overweight and stunting-overweight was 5·3, 10·5, 10·1 and 1·6 %, respectively. In multivariate analyses, children under 6 months, children from low and middle wealth tertiles, and children living in rural areas were prone to being wasted. Male children, low wealth tertile and a non-Thai speaking household head were positively and children aged 48-59 months and a one-child household were inversely associated with stunting. Children from a low wealth tertile were less likely to be overweight, while older age, male children and children from a one-child household were more likely to be overweight. Stunting-overweight was associated with children aged 24-47 months, male children, mothers having secondary education, a one-child household, a non-Thai speaking household head and an urban area. In terms of IYCF indicators, despite no association with stunting and stunted-overweight children, current breast-feeding and inadequate meal frequency were associated with being wasting, while current breast-feeding and dietary diversity were inversely associated with being overweight. CONCLUSIONS: This study revealed the double burden of malnutrition at the individual and population levels among Thai children under-five, which calls for concrete integrated interventions to tackle all forms of malnutrition.

Prevalence and Factors Associated with Postpartum Depression in Primary Healthcare Centres in Yangon, Myanmar.

BACKGROUND: Postpartum depression (PPD) can have serious consequences on both the mother and infant. Despite the higher prevalence, there are limited numbers of studies on PPD in low-and middle-income countries, like Myanmar. This study aimed to explore the prevalence and associated factors of PPD in primary healthcare settings in Myanmar. METHODS: This cross-sectional online study was conducted with 220 mothers under 6 months postpartum in April-May 2020 and who registered in public health centres in Kungyangone Township, Yangon, Myanmar. The postpartum depression was measured with the Edinburgh postpartum depression scale (EPDS, ≥ 13 scores). Independent variables included sociodemographic factors, obstetric and infant factors, psychosocial factors (social support and social media usage), health services utilisation and accessibility factors. Chi-square tests and multiple logistic regression were performed. RESULTS: Overall prevalence of depressive symptoms in 220 women under 6 months postpartum was 31.8% (95% confidence interval [CI]: 25.9, 37.3). In multiple logistic regression, unplanned pregnancy (adjusted odds ratio [AOR]: 2.946), less than four times antenatal care (ANC) visits (AOR: 2.518), travel time more than 1 h to reach health centres (AOR: 3.068) and birth interval more than 5 years (AOR: 4.594) were more likely to be associated with PPD, while preterm delivery (AOR: 0.091) was inversely associated. CONCLUSION: This study showed the relatively high prevalence of PPD and the strong association with preterm delivery, pregnancy intention, breastfeeding status, birth interval as well as frequency of ANC received and travel time to health centre. It may suggest that maternal mental health services should be integrated with existing maternal and child health (MCH) services for early detection and prevention of depression symptoms with promotion of MCH services utilisation and improved accessibility among mothers in primary healthcare setting.

Experimental verification of the value of the Tpeak -Tend interval in ventricular arrhythmia inducibility in an early repolarization syndrome model.

INTRODUCTION: In patients with early repolarization patterns on ECG, many researchers have studied to find predictors of fatal arrhythmia. However, there are no satisfying clinical predictors. We evaluated the value of the Tpeak -Tend interval on pseudo-ECG in canine myocardial wedge preparation models of early repolarization syndrome. METHODS AND RESULTS: Transmural pseudo-ECG and endocardial/epicardial action potentials were recorded from coronary-perfused canine left ventricular wedge preparations (n = 34). The Ito agonist NS5806 (8-10 µM), the calcium channel blocker verapamil (3 µM) and acetylcholine (2-3 µM) were used to mimic the disease model. A ventricular arrhythmia induction test was performed. QTpeak , QTend , Tpeak -Tend , and Tpeak -Tend /QTend were measured at 15 to 20 minutes after the provocative agent infusion. Polymorphic ventricular tachycardias (pVT) developed in 23 of the 34 preparations (67%). The maximal values of Tpeak -Tend and Tpeak -Tend /QTend were recorded just before pVT induction. At baseline, without the provocative agents, Tpeak -Tend and Tpeak -Tend /QTend were not different between pVT-induced and pVT-noninduced preparations. The Tpeak -Tend of the pVT-induced preparations was longer than that of non-induced preparations (58 ± 26.8 msec vs 33 ± 6.8 msec, P < .001). The Tpeak -Tend /QTend of pVT- induced preparations was larger than that of noninduced preparations (0.220 ± 0.1017 vs 0.128 ± 0.0312, P < .001). The transmural and epicardial dispersion of repolarization of pVT-induced preparations were larger than those of pVT-noninduced preparations. The transmural dispersion of repolarization showed a positive correlation with Tpeak -Tend . CONCLUSION: Tpeak -Tend predicted malignant ventricular arrhythmias in early repolarization syndrome models. Tpeak -Tend reflects the repolarization heterogeneity of ventricular myocardium.

Early Adolescent Patterns of Alcohol and Tobacco Use in Eight Association of South-East Asian Nations (ASEAN) Member States.

BACKGROUND: Though alcohol and tobacco are the most commonly used substances among adolescents, little is known about the patterning of early adolescent substance use in Association of South-East Asian Nations (ASEAN) member states. OBJECTIVES: This study examined past month patterns of substances use and its gender difference among adolescents. METHODS: Cross-sectional samples among adolescents aged 13-16 years who completed the Global School-based Student Health Survey (GSHS) from eight ASEAN countries were included in the analysis (n = 40,212). RESULTS: Prevalence of past month any tobacco use was relatively high in Brunei, Indonesia, Malaysia, Thailand, and the Philippines (11-15%), but prevalence of dual cigarette and other forms of tobacco use was about 2-5% in the five countries. Past month alcohol consumption prevalence was also high in Thailand, Viet Nam, and the Philippines (16-24%), compared to the rest countries (1.4-8.2%). Moreover, prevalence of the concurrent alcohol and tobacco use was higher in Thailand and the Philippines (7 and 10%, respectively), particularly in boys (13 and 15%, respectively). Conclusions/importance: Almost 30-40% of the boys and 10-20% of girls in Malaysia, Philippines, Thailand, and Viet Nam are engaged in at least one of the two risk behaviors, and the concurrent alcohol and tobacco use was also relatively high among boys in those countries (5-15%). This study may provide some valuable insights on alcohol and tobacco policy in the region and requires to begin prevention and treatment programs in ASEAN member states.

Parental misperception of child's weight and related factors within family norms.

PURPOSE: Parental perception of their child's weight may be a crucial factor in parental ability for action with regard to their child's weight problem. This aim of this study was to investigate parental perception of their child's weight status and dietary healthiness, amount of food consumed and physical activity level and its related factors. METHODS: A cross-sectional survey was conducted among children (Grades 4-6) selected by cluster sampling in two schools. Children were invited to participate in the measurements of anthropometry and their parents were asked to classify their child's weight and health behaviors. RESULTS: In total, 41.8% of parents misperceived their child's weight, of which 82% underestimated their child's weight, in particular regarding overweight or obesity. As parents of overweight or obese children underestimated their child's weight, around 65% were not concerned with their child's current weight and about becoming overweight in the future. Factor associated with underestimation of overweight children was not having a sibling, while among children with normal weight, the underestimation was associated with boys, lower body mass index (BMI), maternal employment and low household income. Furthermore, parents underestimating their child's weight were more likely to be optimistic about their child's dietary healthiness, food amount taken, and physical activity level than those with correct child's weight estimates. CONCLUSIONS: Findings show a high proportion of parental misperception of their child's weight status. Family-based weight control interventions will need to incorporate parental misperceptions of the body weight and health behaviors of their children.

Quantification and tracking of genetically engineered dendritic cells for studying immunotherapy.

PURPOSE: Genetically encoded reporters can assist in visualizing biological processes in live organisms and have been proposed for longitudinal and noninvasive tracking of therapeutic cells in deep tissue. Cells can be labeled in situ or ex vivo and followed in live subjects over time. Nevertheless, a major challenge for reporter systems is to identify the cell population that actually expresses an active reporter. METHODS: We have used a nucleoside analog, pyrrolo-2'-deoxycytidine, as an imaging probe for the putative reporter gene, Drosophila melanogaster 2'-deoxynucleoside kinase. Bioengineered cells were imaged in vivo in animal models of brain tumor and immunotherapy using chemical exchange saturation transfer MRI. The number of transduced cells was quantified by flow cytometry based on the optical properties of the probe. RESULTS: We performed a comparative analysis of six different cell lines and demonstrate utility in a mouse model of immunotherapy. The proposed technology can be used to quantify the number of labeled cells in a given region, and moreover is sensitive enough to detect less than 10,000 cells. CONCLUSION: This unique technology that enables efficient selection of labeled cells followed by in vivo monitoring with both optical and MRI. Magn Reson Med 79:1010-1019, 2018. © 2017 International Society for Magnetic Resonance in Medicine.

Biodegradable Polymeric Nanocarrier-Based Immunotherapy in Hepatitis Vaccination.

Various commercial vaccines are used for immunization against hepatitis B. However, these immunotherapeutic vaccines require invasive administration, which can induce side effects, and require multiple shots to elicit an immune response, limiting their efficacy. Compared to traditional hepatitis B vaccines, polymer nanoparticles have more advantageous inherent properties as vaccine delivery carriers, providing increased stability of encapsulated antigen, the possibility of single-shot immunotherapy, and the capability of mucosal administration, which allows various routes of vaccination. In this review, we present up-to-date information on the potential of a biodegradable nanoparticle-based delivery system in treating hepatitis B. We also discuss the application of nanoparticles in various vaccines and highlighted strategies for eliciting an appropriate immune response.