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, , Sherry G. Mansour, , Jason H. Greenberg, Yu Yamamoto, Aditya Biswas,
Published: 12 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251376

Abstract:
Importance False negative SARS-CoV-2 tests can lead to spread of infection in the inpatient setting to other patients and healthcare workers. However, the population of patients with COVID who are admitted with false negative testing is unstudied. Objective To characterize and develop a model to predict true SARS-CoV-2 infection among patients who initially test negative for COVID by PCR. Design Retrospective cohort study. Setting Five hospitals within the Yale New Haven Health System between 3/10/2020 and 9/1/2020. Participants Adult patients who received diagnostic testing for SARS-CoV-2 virus within the first 96 hours of hospitalization. Exposure We developed a logistic regression model from readily available electronic health record data to predict SARS-CoV-2 positivity in patients who were positive for COVID and those who were negative and never retested. Main outcomes and measures This model was applied to patients testing negative for SARS-CoV-2 who were retested within the first 96 hours of hospitalization. We evaluated the ability of the model to discriminate between patients who would subsequently retest negative and those who would subsequently retest positive. Results We included 31,459 hospitalized adult patients; 2,666 of these patients tested positive for COVID and 3,511 initially tested negative for COVID and were retested. Of the patients who were retested, 61 (1.7%) had a subsequent positive COVID test. The model showed that higher age, vital sign abnormalities, and lower white blood cell count served as strong predictors for COVID positivity in these patients. The model had moderate performance to predict which patients would retest positive with a test set area under the receiver-operator characteristic (ROC) of 0.76 (95% CI 0.70–0.83). Using a cutpoint for our risk prediction model at the 90th percentile for probability, we were able to capture 35/61 (57%) of the patients who would retest positive. This cutpoint amounts to a number-needed-to-retest range between 15 and 77 patients. Conclusion and relevance We show that a pragmatic model can predict which patients should be retested for COVID. Further research is required to determine if this risk model can be applied prospectively in hospitalized patients to prevent the spread of SARS-CoV-2 infections.
, Dhanasekaran Bodhini, Juanjie Jiang, Kandaswamy Ramya, Deepa Mohan, Coimbatore Subramanian Shanthi Rani, Nagarajan Lakshmipriya, Vasudevan Sudha, Rajendra Pradeepa, Ranjit Mohan Anjana, et al.
Published: 12 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0238555

Abstract:
Background Plasma omentin levels have been shown to be associated with circulating adiponectin concentrations and cardiometabolic disease-related outcomes. In this study, we aim to examine the association of omentin gene polymorphism with serum adiponectin levels and cardiometabolic health status using a genetic approach, and investigate whether these associations are modified by lifestyle factors. Methods The study included 945 normal glucose tolerant and 941 unrelated individuals with type 2 diabetes randomly selected from the Chennai Urban Rural Epidemiology Study (CURES), in southern India. Study participants were classified into cardiometabolically healthy and unhealthy, where cardiometabolically healthy were those without hypertension, diabetes, and dyslipidemia. Fasting serum adiponectin levels were measured by radioimmunoassay. The omentin A326T (rs2274907) single nucleotide polymorphism (SNP) was screened by polymerase chain reaction-restriction fragment length polymorphism and direct sequencing. Results The ‘A’ allele of the omentin SNP was significantly associated with lower adiponectin concentrations after adjusting for age, sex, body mass index (BMI), waist circumference (WC) and cardiometabolic health status (p = 1.90 x 10−47). There was also a significant association between circulating adiponectin concentrations and cardiometabolic health status after adjusting for age, sex, BMI, WC and Omentin SNP (p = 7.47x10-10). However, after adjusting for age, sex, BMI, WC and adiponectin levels, the association of ‘A’ allele with cardiometabolic health status disappeared (p = 0.79) suggesting that adiponectin serves as a mediator of the association between omentin SNP and cardiometabolic health status. There were no significant interactions between the SNP and dietary factors on adiponectin levels and cardiometabolic health status (p>0.25, for all comparisons). Conclusions Our findings show that adiponectin might function as a mechanistic link between omentin SNP and increased risk of cardiometabolic diseases independent of common and central obesity in Asian Indians. Before strategies to promote adiponectin modulation could be implemented, further studies are required to confirm the molecular mechanisms involved in this triangular relationship between omentin gene, adiponectin and cardiometabolic diseases.
, Mitchell Wong, Kulwant Dosanjh, Christopher Biely, Rebecca Dudovitz
Published: 12 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251332

Abstract:
Objectives Although racial stigma in school is associated with adolescent risky health behaviors, there are no studies investigating how gender stigma relates to adolescent risky health behaviors among low-income, minority youth. We sought to determine whether gender stigma awareness is associated with adolescent risky health behaviors (delinquency, fighting, and substance use) and whether this association is mediated by school disengagement (low perceived teacher support, low school engagement, cutting classes, and breaking school rules) among low-income, minority students. Methods We analyzed cross-sectional survey data, collected from 2017 to 2019, from 412 high school students. Multi-level logistic regressions tested whether gender stigma awareness was associated with delinquency, fighting, and substance use, controlling for covariates, baseline behaviors, and clustering within schools. Mediation analyses tested whether school disengagement (low school engagement, perceived teacher support, cutting class, and breaking school rules) mediated these associations. Secondary analyses explored whether associations differed for male versus female, high-performing versus low-performing, and Latinx versus non-Latinx students. Results In this predominantly Latinx (83%) sample, gender stigma awareness was associated with delinquency (AOR = 1.48, P< 0.001) and fighting (AOR = 1.15, P< 0.001). School engagement, perceived teacher support, breaking school rules, and cutting classes mediated 42.7% of the association between gender stigma awareness and delinquency and 65.42% of the association between gender stigma awareness and fighting. Gender stigma awareness was also associated with substance use for low-performing (AOR = 1.68, P = 0.003) and non-Latinx adolescents (AOR = 3.80, P = 0.03). School disengagement did not mediate the association between gender stigma awareness and substance use for non-Latinx students but mediated 50% of this association for low-performing students. Conclusions Gender stigma awareness is associated with adolescent risky health behaviors. A decreased sense of acceptance in the school community and increased school misbehavior may mediate these associations. School environments that value and accept all students may better support adolescent health.
Correction
The Plos One Staff
Published: 11 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251757

Mohamad Malik Al-Adil Baharudin, , Fairolniza Mohd Shariff, Zetty Norhana Balia Yusof, Murni Karim, Syarul Nataqain Baharum,
Published: 11 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251514

Abstract:
Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) have reached epidemic proportions globally. Therefore, there is an urgent need for a continuous supply of antibiotics to combat the problem. In this study, bacteria initially identified as species belonging to the Bacillus amyloliquefaciens operational group were re-identified based on the housekeeping gene, gyrB. Cell-free supernatants (CFS) from the strains were used for antimicrobial tests using the agar well diffusion assay against MRSA and various types of pathogenic bacteria. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and physicochemical characteristics of the CFS were determined. Based on gyrB sequence analysis, five strains (PD9, B7, PU1, BP1 and L9) were identified as Bacillus velezensis. The CFS of all B. velezensis strains showed broad inhibitory activities against Gram-negative and -positive as well as MRSA strains. Strain PD9 against MRSA ATCC 33742 was chosen for further analysis as it showed the biggest zone of inhibition (21.0 ± 0.4 mm). The MIC and MBC values obtained were 125 μl/ml. The crude antimicrobial extract showed bactericidal activity and was stable at various temperatures (40–80°C), pH (4–12), surfactants (Tween 20, Tween 80, SDS and Triton X-100) and metal ions (MgCI2, NaCI2, ZnNO3 and CuSO4) when tested. However, the crude extract was not stable when treated with proteinase K. All these properties resembled the characteristics of peptides. The antimicrobial compound from the selected strain was purified by using solvent extraction method and silica gel column chromatography. The purified compound was subjected to High Performance Liquid Chromatography which resulted in a single peak of the anti-MRSA compound being detected. The molecular weight of the anti-MRSA compound was determined by using SDS-PAGE and zymogram. The size of the purified antimicrobial peptide was approximately ~ 5 kDa. The antimicrobial peptide produced from B. velezensis strain PD9 is a promising alternative to combat the spread of MRSA infections in the future.
Zhaofang Chen, Kexin Shi, Wenjie Kuang,
Published: 11 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0250885

Abstract:
Cadmium (Cd) is a toxic non-essential element, while calcium (Ca) is an essential element with high chemical similarity to Cd. Dietary intake is the major Cd exposure pathway for non-smokers. A multi-concentration dietary intervention experiment was designed to explore the optimum concentration of Ca in diet with obvious protective effects against the toxicity of livers and kidneys induced by Cd in mice. The mice were divided into six groups with different concentrations of Cd and Ca in their food: control-group (no Cd or Ca), Ca-group (100 g/kg Ca, without Cd), Cd-group (2 mg/kg Cd, without Ca), CaL+Cd-group (2 mg/kg Cd, 2 g/kg Ca), CaM+Cd-group (2 mg/kg Cd, 20 g/kg Ca) and CaH+Cd-group (2 mg/kg Cd, 100 g/kg Ca). The organ indexes, oxidative stress biomarkers, lesions and Cd concentrations were detected after a 30-day exposure period. Results showed that serum Aspartate Aminotransferase (AST) level in CaH+Cd-group was significantly lower than that in Cd-group, while close to that in control-group. The contents of Serum Blood Urea Nitrogen (BUN) in different groups showed the same trend. Concentrations of all oxidative stress biomarkers (GSH-Px, SOD, CAT, GSH and MDA) in CaH+Cd-group were close to the normal levels of control-group while significantly different from those in Cd-group. The only exception was the Malondialdehyde (MDA) levels in kidneys. This study suggests that Ca plays a protective role in relieving the Cd-induced toxicity of livers and kidneys and a concentration of 100 g/kg for Ca in diet showed the best protective effects. These findings could provide a clue for further studies concerning human diet intervention for Cd control.
Guangzhi Yang, BoShen Wang, Dawei Sun, Huimin Wang, Mengyao Chen, Hao Chen,
Published: 11 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251090

Abstract:
Noise-induced-hearing-loss(NIHL) is a common occupational disease caused by various environmental and biological factors. To investigate the association between TAB2 and the susceptibility of NIHL of people exposed to occupational environments, a genetic association study was performed on selected companies with 588 cases and 537 healthy control subjects. Five selected single nucleotide polymorphisms (SNPs) in TAB2,incoluding rs2744434, rs521845, rs652921, rs7896, rs9485372, were genotyped after a collection of DNA samples. Evident differences in participants between the case group and the control group reveals the result that people with the TAB2 has a high probability of getting NIHL. The results show that rs521845 is deeply associated with the risk of NIHL and is available for the diagnosis in the future.
, David Armisén, Séverine Viala, Amélie Decaras,
PLOS Biology, Volume 19; doi:10.1371/journal.pbio.3001157

Abstract:
Exaggerated sexually selected traits, often carried by males, are characterized by the evolution of hyperallometry, resulting in their disproportionate growth relative to the rest of the body among individuals of the same population. While the evolution of allometry has attracted much attention for centuries, our understanding of the developmental genetic mechanisms underlying its emergence remains fragmented. Here we conduct comparative transcriptomics of the legs followed by an RNA interference (RNAi) screen to identify genes that play a role in the hyperallometric growth of the third legs in the males of the water strider Microvelia longipes. We demonstrate that a broadly expressed growth factor, Bone Morphogenetic Protein 11 (BMP11, also known as Growth Differentiation Factor 11), regulates leg allometries through increasing the allometric slope and mean body size in males. In contrast, BMP11 RNAi reduced mean body size but did not affect slope either in the females of M. longipes or in the males and females of other closely related Microvelia species. Furthermore, our data show that a tissue-specific factor, Ultrabithorax (Ubx), increases intercept without affecting mean body size. This indicates a genetic correlation between mean body size and variation in allometric slope, but not intercept. Strikingly, males treated with BMP11 RNAi exhibited a severe reduction in fighting frequency compared to both controls and Ubx RNAi-treated males. Therefore, male body size, the exaggerated weapon, and the intense fighting behavior associated with it are genetically correlated in M. longipes. Our results support a possible role of pleiotropy in the evolution of allometric slope.
, Jean-Baptiste Pavani, Javier Fernandez-Alvarez, Azucena Garcia-Palacios, Cristina Botella
Published: 11 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251561

Abstract:
A growing body of research has investigated the regulation of negative emotions in ecological settings, but little is known about the mechanisms underlying positive emotion regulation in everyday life. Although some evidence suggests that adopting positive strategies is beneficial for emotional well-being, the literature is inconsistent about the effects of positive emotions on subsequent regulatory processes. In the present study, we adopted a two-week ecological momentary assessment to explore the association between positive emotions and positive emotion regulation in daily life. According to our results, the less individuals felt positive emotions at one point, the more they tended to enhance their use of positive strategies from this time to the next, which in turn resulted in subsequent higher levels of positive emotions. This prototype of positive regulation can be seen as a highly adaptive mechanism that makes it possible to compensate for a lack of positive emotions by enhancing the deployment of positive strategies. The theoretical and clinical implications of these findings are discussed.
, , , Rebecca Nzawa, Richard Chilongosi, , Rose Nyirenda, , , Rachel Baggaley, et al.
PLOS Medicine, Volume 18; doi:10.1371/journal.pmed.1003608

Abstract:
Background Undiagnosed HIV infection remains substantial in key population subgroups including adolescents, older adults, and men, driving ongoing transmission in sub-Saharan Africa. We evaluated the impact, safety, and costs of community-led delivery of HIV self-testing (HIVST), aiming to increase HIV testing in underserved subgroups and stimulate demand for antiretroviral therapy (ART). Methods and findings This cluster-randomised trial, conducted between October 2018 and July 2019, used restricted randomisation (1:1) to allocate 30 group village head clusters in Mangochi district, Malawi to the community-led HIVST intervention in addition to the standard of care (SOC) or the SOC alone. The intervention involved mobilising community health groups to lead the design and implementation of 7-day HIVST campaigns, with cluster residents (≥15 years) eligible for HIVST. The primary outcome compared lifetime HIV testing among adolescents (15 to 19 years) between arms. Secondary outcomes compared: recent HIV testing (in the last 3 months) among older adults (≥40 years) and men; cumulative 6-month incidence of ART initiation per 100,000 population; knowledge of the preventive benefits of HIV treatment; and HIV testing stigma. Outcomes were measured through a post-intervention survey and at neighboring health facilities. Analysis used intention-to-treat for cluster-level outcomes. Community health groups delivered 24,316 oral fluid-based HIVST kits. The survey included 90.2% (3,960/4,388) of listed participants in the 15 community-led HIVST clusters and 89.2% (3,920/4,394) of listed participants in the 15 SOC clusters. Overall, the proportion of men was 39.0% (3,072/7,880). Most participants obtained primary-level education or below, were married, and reported a sexual partner. Lifetime HIV testing among adolescents was higher in the community-led HIVST arm (84.6%, 770/910) than the SOC arm (67.1%, 582/867; adjusted risk difference [RD] 15.2%, 95% CI 7.5% to 22.9%; p < 0.001), especially among 15 to 17 year olds and boys. Recent testing among older adults was also higher in the community-led HIVST arm (74.5%, 869/1,166) than the SOC arm (31.5%, 350/1,111; adjusted RD 42.1%, 95% CI 34.9% to 49.4%; p < 0.001). Similarly, the proportions of recently tested men were 74.6% (1,177/1,577) and 33.9% (507/1,495) in the community-led HIVST and SOC arms, respectively (adjusted RD 40.2%, 95% CI 32.9% to 47.4%; p < 0.001). Knowledge of HIV treatment benefits and HIV testing stigma showed no differences between arms. Cumulative incidence of ART initiation was respectively 305.3 and 226.1 per 100,000 population in the community-led HIVST and SOC arms (RD 72.3, 95% CI −36.2 to 180.8; p = 0.18). In post hoc analysis, ART initiations in the 3-month post-intervention period were higher in the community-led HIVST arm than the SOC arm (RD 97.7, 95% CI 33.4 to 162.1; p = 0.004). HIVST uptake was 74.7% (2,956/3,960), with few adverse events (0.6%, 18/2,955) and at US$5.70 per HIVST kit distributed. The main limitations include the use of self-reported HIV testing outcomes and lack of baseline measurement for the primary outcome. Conclusions In this study, we found that community-led HIVST was effective, safe, and affordable, with population impact and coverage rapidly realised at low cost. This approach could enable community HIV testing in high HIV prevalence settings and demonstrates potential for economies of scale and scope. Trial registration Clinicaltrials.gov NCT03541382.
Takuro Kobori, Mayuka Tameishi, Chihiro Tanaka, Yoko Urashima, Tokio Obata
Published: 11 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0250889

Abstract:
The ezrin/radixin/moesin (ERM) family proteins act as linkers between the actin cytoskeleton and P-glycoprotein (P-gp) and regulate the plasma membrane localization and functionality of the latter in various cancer cells. Notably, P-gp overexpression in the plasma membrane of cancer cells is a principal factor responsible for multidrug resistance and drug-induced mutagenesis. However, it remains unknown whether the ERM proteins contribute to the plasma membrane localization and transport function of P-gp in human colorectal cancer cells in which the subcellular localization of ERM has yet to be determined. This study aimed to determine the gene expression patterns and subcellular localization of ERM and P-gp and investigate the role of ERM proteins in the plasma membrane localization and transport function of P-gp using the human colon adenocarcinoma cell line LS180. Using real-time reverse transcription polymerase chain reaction and immunofluorescence analyses, we showed higher levels of ezrin and moesin mRNAs than those of radixin mRNA in these cells and preferential distribution of all three ERM proteins on the plasma membrane. The ERM proteins were highly colocalized with P-gp. Additionally, we show that the knockdown of ezrin, but not of radixin and moesin, by RNA interference significantly decreased the cell surface expression of P-gp in LS180 cells without affecting the mRNA expression of P-gp. Furthermore, gene silencing of ezrin substantially increased the intracellular accumulation of rhodamine123, a typical P-gp substrate, with no alterations in the plasma membrane permeability of Evans blue, a passive transport marker. In conclusion, ezrin may primarily regulate the cell surface localization and transport function of P-gp as a scaffold protein without influencing the transcriptional activity of P-gp in LS180 cells. These findings should be relevant for treating colorectal cancer, which is the second leading cause of cancer-related deaths in males and females combined.
Emmanuel Echeverry-Cárdenas, Carolina López-Castañeda, ,
PLOS Neglected Tropical Diseases, Volume 15; doi:10.1371/journal.pntd.0008212

Abstract:
In Colombia, little is known on the distribution of the Asian mosquito Aedes albopictus, main vector of dengue, chikungunya, and Zika in Asia and Oceania. Therefore, this work sought to estimate its current and future potential geographic distribution under the Representative Concentration Paths (RCP) 2.6 and 8.5 emission scenarios by 2050 and 2070, using ecological niche models. For this, predictions were made in MaxEnt, employing occurrences of A. albopictus from their native area and South America and bioclimatic variables of these places. We found that, from their invasion of Colombia to the most recent years, A. albopictus is present in 47% of the country, in peri-urban (20%), rural (23%), and urban (57%) areas between 0 and 1800 m, with Antioquia and Valle del Cauca being the departments with most of the records. Our ecological niche modelling for the currently suggests that A. albopictus is distributed in 96% of the Colombian continental surface up to 3000 m (p < 0.001) putting at risk at least 48 million of people that could be infected by the arboviruses that this species transmits. Additionally, by 2050 and 2070, under RCP 2.6 scenario, its distribution could cover to nearly 90% of continental extension up to 3100 m (≈55 million of people at risk), while under RCP 8.5 scenario, it could decrease below 60% of continental extension, but expand upward to 3200 m (< 38 million of people at risk). These results suggest that, currently in Colombia, A. albopictus is found throughout the country and climate change could diminish eventually its area of distribution, but increase its altitudinal range. In Colombia, surveillance and vector control programs must focus their attention on this vector to avoid complications in the national public health setting.
Kennedy Edemacu, ,
Published: 11 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0250992

Abstract:
With the rapid advancement of information and communication technologies, there is a growing transformation of healthcare systems. A patient’s health data can now be centrally stored in the cloud and be shared with multiple healthcare stakeholders, enabling the patient to be collaboratively treated by more than one healthcare institution. However, several issues, including data security and privacy concerns still remain unresolved. Ciphertext-policy attribute-based encryption (CP-ABE) has shown promising potential in providing data security and privacy in cloud-based systems. Nevertheless, the conventional CP-ABE scheme is inadequate for direct adoption in a collaborative ehealth system. For one, its expressiveness is limited as it is based on a monotonic access structure. Second, it lacks an attribute/user revocation mechanism. Third, the computational burden on both the data owner and data users is linear with the number of attributes in the ciphertext. To address these inadequacies, we propose CESCR, a CP-ABE for efficient and secure sharing of health data in collaborative ehealth systems with immediate and efficient attribute/user revocation. The CESCR scheme is unbounded, i.e., it does not bind the size of the attribute universe to the security parameter, it is based on the expressive and non-restrictive ordered binary decision diagram (OBDD) access structure, and it securely outsources the computationally demanding attribute operations of both encryption and decryption processes without requiring a dummy attribute. Security analysis shows that the CESCR scheme is secure in the selective model. Simulation and performance comparisons with related schemes also demonstrate that the CESCR scheme is expressive and efficient.
Sarah Sternbach, Nicole West, Naveen K. Singhal, Robert Clements, Soumitra Basu, Ajai Tripathi, Ranjan Dutta, Ernest J. Freeman,
Published: 11 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0250486

Abstract:
Research into the epigenome is of growing importance as a loss of epigenetic control has been implicated in the development of neurodegenerative diseases. Previous studies have implicated aberrant DNA and histone methylation in multiple sclerosis (MS) disease pathogenesis. We have previously reported that the methyl donor betaine is depleted in MS and is linked to changes in histone H3 trimethylation (H3K4me3) in neurons. We have also shown that betaine increases histone methyltransferase activity by activating chromatin bound betaine homocysteine S-methyltransferase (BHMT). Here, we investigated the role of the BHMT-betaine methylation pathway in oligodendrocytes. Immunocytochemistry in the human MO3.13 cell line, primary rat oligodendrocytes, and tissue from MS postmortem brain confirmed the presence of the BHMT enzyme in the nucleus in oligodendrocytes. BHMT expression is increased 2-fold following oxidative insult, and qRT-PCR demonstrated that betaine can promote an increase in expression of oligodendrocyte maturation genes SOX10 and NKX-2.2 under oxidative conditions. Chromatin fractionation provided evidence of a direct interaction of BHMT on chromatin and co-IP analysis indicates an interaction between BHMT and DNMT3a. Our data show that both histone and DNA methyltransferase activity are increased following betaine administration. Betaine effects were shown to be dependent on BHMT expression following siRNA knockdown of BHMT. This is the first report of BHMT expression in oligodendrocytes and suggests that betaine acts through BHMT to modulate histone and DNA methyltransferase activity on chromatin. These data suggest that methyl donor availability can impact epigenetic changes and maturation in oligodendrocytes.
, Micaela Bianchi, Catalina von Bilderling,
Published: 11 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251411

Abstract:
Cells exert traction forces on the extracellular matrix to which they are adhered through the formation of focal adhesions. Spatial-temporal regulation of traction forces is crucial in cell adhesion, migration, cellular division, and remodeling of the extracellular matrix. By cultivating cells on polyacrylamide hydrogels of different stiffness we were able to investigate the effects of substrate stiffness on the generation of cellular traction forces by Traction Force Microscopy (TFM), and characterize the molecular dynamics of the focal adhesion protein zyxin by Fluorescence Correlation Spectroscopy (FCS) and Fluorescence Recovery After Photobleaching (FRAP). As the rigidity of the substrate increases, we observed an increment of both, cellular traction generation and zyxin residence time at the focal adhesions, while its diffusion would not be altered. Moreover, we found a positive correlation between the traction forces exerted by cells and the residence time of zyxin at the substrate elasticities studied. We found that this correlation persists at the subcellular level, even if there is no variation in substrate stiffness, revealing that focal adhesions that exert greater traction present longer residence time for zyxin, i.e., zyxin protein has less probability to dissociate from the focal adhesion.
Dini Hu, Yuzhu Chao, Boru Zhang, Chen Wang, Yingjie Qi, Make Ente, Dong Zhang, Kai Li,
Published: 11 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251512

Abstract:
Horse botflies have been a threat to the Przewalski’s horses in the Kalamaili Nature Reserve in Xinjiang of China since their reintroduction to the original range. As larvae of these parasites could infest the intestine of a horse for months, they could interact with and alter the structure and composition of its intestinal microbiota, affecting adversely its health. Nonetheless, there are no such studies on the rewilded Przewalski’s horses yet. For the first time, this study characterizes the composition of the intestinal microbiota of 7 rewilded Przewalski’s horses infected severely by Gasterophilus pecorum following and prior to their anthelmintic treatment. Bioinformatics analyses of the sequence data obtained by amplicon high throughput sequencing of bacterial 16S rRNA genes showed that G. pecorum infestation significantly increased the richness of the intestinal microbial community but not its diversity. Firmicutes and Bacteroidetes were found the dominant phyla as in other animals, and the parasitic infestation decreased the F/B ratio largely by over 50%. Large reduction in relative abundances of the two genera Streptococcus and Lactobacillus observed with G. pecorum infestation suggested possible changes in colic and digestion related conditions of the infected horses. Variations on the relative abundance of the genus groups known to be pathogenic or symbiotic showed that adverse impact of the G. pecorum infestation could be associated with reduction of the symbiotic genera Lactobacillus and Bifidobacterium that are probiotics and able to promote immunity against parasitic infection.
Nisha Venugopal,
Published: 11 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251191

Abstract:
Background It is an ethical and scientific obligation to register each clinical trial, and report its results, accurately, comprehensively and on time. The WHO recognizes 17 public registries as Primary Registries, and has also introduced a set of minimal standards in the International Standards for Clinical Trial Registries (ISCTR) that primary registries need to implement. These standards are categorized into nine sections—Content, Quality and Validity, Accessibility, Unambiguous Identification, Technical Capacity, Administration and Governance, the Trial Registration Data Set (TRDS), Partner registries and Data Interchange Standards. This study compared the WHO’s primary registries, and the US’s ClinicalTrials.gov, to examine the implementation of ISCTR, with the aim of defining features of an interim ideal registry. Methods and findings The websites of the 18 registries were evaluated for 14 features that map to one or more of the nine sections of ISCTR, and assigned scores for their variations of these features. The assessed features include the nature of the content; the number and nature of fields to conduct a search; data download formats; the nature of the audit trail; the health condition category; the documentation available on a registry website; etc. The registries received scores for their particular variation of a given feature based on a scoring rationale devised for each individual feature analysed. Overall, the registries received between 27% and 80% of the maximum score of 94. The results from our analysis were used to define a set of features of an interim ideal registry. Conclusions To the best of our knowledge, this is the first study to quantify the widely divergent quality of the primary registries’ compliance with the ISCTR. Even with this limited assessment, it is clear that some of the registries have much work to do, although even a few improvements would significantly improve them.
Correction
Monica Viegas Andrade, Augusto Quaresma Coelho, Mauro Xavier Neto, Lucas Resende de Carvalho, Rifat Atun, Marcia C. Castro
Published: 11 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251764

Farrukh Jamal, Christophe Chesneau, Dalal Lala Bouali,
Published: 11 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0250790

Abstract:
In recent years, the trigonometric families of continuous distributions have found a place of choice in the theory and practice of statistics, with the Sin-G family as leader. In this paper, we provide some contributions to the subject by introducing a flexible extension of the Sin-G family, called the transformed Sin-G family. It is constructed from a new polynomial-trigonometric function presenting a desirable “versatile concave/convex” property, among others. The modelling possibilities of the former Sin-G family are thus multiplied. This potential is also highlighted by a complete theoretical work, showing stochastic ordering results, studying the analytical properties of the main functions, deriving several kinds of moments, and discussing the reliability parameter as well. Then, the applied side of the proposed family is investigated, with numerical results and applications on the related models. In particular, the estimation of the unknown model parameters is performed through the use of the maximum likelihood method. Then, two real life data sets are analyzed by a new extended Weibull model derived to the considered trigonometric mechanism. We show that it performs the best among seven comparable models, illustrating the importance of the findings.
, Carlos Martínez Hernández, María Luisa Rico Gómez
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251398

Abstract:
Heritage and space establish reciprocal relations that have been studied for decades. On the one hand, heritage has been described as an inherently spatial phenomenon. On the other hand, places are defined according to the attributes that make up their identity, among which heritage is a fundamental instrument. On the basis of the idea that education plays an important role in the socialization process, transmitted by the inherited culture, to integrate each subject within the specific community, and the notion of scale as the closest to heritage, we defined as general objectives to determine the relationships between geographic scales, heritage perspective and the didactic potential granted to heritage, within the framework of the construction of collective identities, and to contrast the perspectives of students and teachers regarding the geographical scale, heritage and their didactic potential, deducing implications for educational practices. In order to answer to these objectives, we carried out a non-experimental quantitative research, with a relational-predictive objective. Specifically, we used a survey method, being the context the whole of the local scale (Fuente Álamo, Murcia, Spain) and acting as participants all students and teachers of Secondary Education (n = 459) linked to social sciences. They answered the Test on Didactic Potentiality of Heritage according to Scale (TDPHS), and its information was analysed through different procedures (Spearman’s correlations, descriptive statistics, Mann-Whitney U…), using the statistical programs SPSS. The results show, on the one hand, that the scalar perspective scores are generally low, heritage perspective is consistent with the consideration of the scales, and the perceived didactic potential in relation to heritage is related to the importance given to each of the scales; and, on the other hand, the contrast in the perspectives of students and teachers regarding the geographical scale, heritage and their didactic potential is minimal.
, , Ana Tereza R. Vasoncelos, Cecilia Hedin-Pereira,
PLOS Computational Biology, Volume 17; doi:10.1371/journal.pcbi.1008956

Abstract:
A major factor contributing to the etiology of depression is a neurochemical imbalance of the dopaminergic and serotonergic systems, which is caused by persistently high levels of circulating stress hormones. Here, a computational model is proposed to investigate the interplay between dopaminergic and serotonergic-kynurenine metabolism under cortisolemia and its consequences for the onset of depression. The model was formulated as a set of nonlinear ordinary differential equations represented with power-law functions. Parameter values were obtained from experimental data reported in the literature, biological databases, and other general information, and subsequently fine-tuned through optimization. Model simulations predict that changes in the kynurenine pathway, caused by elevated levels of cortisol, can increase the risk of neurotoxicity and lead to increased levels of 3,4-dihydroxyphenylaceltahyde (DOPAL) and 5-hydroxyindoleacetaldehyde (5-HIAL). These aldehydes contribute to alpha-synuclein aggregation and may cause mitochondrial fragmentation. Further model analysis demonstrated that the inhibition of both serotonin transport and kynurenine-3-monooxygenase decreased the levels of DOPAL and 5-HIAL and the neurotoxic risk often associated with depression. The mathematical model was also able to predict a novel role of the dopamine and serotonin metabolites DOPAL and 5-HIAL in the ethiology of depression, which is facilitated through increased cortisol levels. Finally, the model analysis suggests treatment with a combination of inhibitors of serotonin transport and kynurenine-3-monooxygenase as a potentially effective pharmacological strategy to revert the slow-down in monoamine neurotransmission that is often triggered by inflammation.
Kristen N. Howard, Emma K. Stapleton, April A. Nelms, Kelsee C. Ryan,
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251275

Abstract:
Reading primary literature is a popular classroom practice that exposes students to the process of science. However, the analysis of primary literature can be taxing and time-consuming for students. For this reason, it is important to determine the source of student challenges and what motivates them to read primary literature. To better understand students’ challenges, preferences, and motivations towards analyzing primary literature, we held focus groups with biology undergraduates where we asked them about their thoughts and perceptions on this practice. Students felt they struggle with understanding the big picture of an article, certain aspects of scientific literacy like data interpretation and experimental setup, and lack of knowledge of terms and techniques. Further analysis of the data using the achievement goal and expectancy-value theories of motivation revealed that students: 1) demonstrate mastery and performance approach goal orientations, which are typically associated with positive learning outcomes, 2) value the usefulness of reading primary literature, and 3) feel most engaged in the process of reading an article when the topic interests them. We provide pedagogical recommendations based on our findings.
, Rostislav Khlebnikov, Alex Melville, Marija Marčan, , , Federica Cuomo, Miguel Silva Vieira, , , et al.
PLOS Computational Biology, Volume 17; doi:10.1371/journal.pcbi.1008881

Abstract:
In this work, we describe the CRIMSON (CardiovasculaR Integrated Modelling and SimulatiON) software environment. CRIMSON provides a powerful, customizable and user-friendly system for performing three-dimensional and reduced-order computational haemodynamics studies via a pipeline which involves: 1) segmenting vascular structures from medical images; 2) constructing analytic arterial and venous geometric models; 3) performing finite element mesh generation; 4) designing, and 5) applying boundary conditions; 6) running incompressible Navier-Stokes simulations of blood flow with fluid-structure interaction capabilities; and 7) post-processing and visualizing the results, including velocity, pressure and wall shear stress fields. A key aim of CRIMSON is to create a software environment that makes powerful computational haemodynamics tools accessible to a wide audience, including clinicians and students, both within our research laboratories and throughout the community. The overall philosophy is to leverage best-in-class open source standards for medical image processing, parallel flow computation, geometric solid modelling, data assimilation, and mesh generation. It is actively used by researchers in Europe, North and South America, Asia, and Australia. It has been applied to numerous clinical problems; we illustrate applications of CRIMSON to real-world problems using examples ranging from pre-operative surgical planning to medical device design optimization.
Lyudmila Maksanova, Taisiya Bardakhanova, Natalia Lubsanova, Darima Budaeva, Arnold Tulokhonov
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251383

Abstract:
The impact of protected areas on local communities is the subject of intense discussions as part of the implementation of the global ecosystem protection agenda. Conflicts between the interests of environmental protection and the needs of socio-economic development become particularly acute when large areas of land are taken out of economic circulation as a result of organizing protected areas. In this case, there is an urgent need for detailed and reliable information about the social impacts of such land withdrawal on the well-being of the local population. An analysis of the methodological approaches widely presented in the literature, used to assess the social impact of protected areas, testifies to the insufficiency of completed and practically applicable methodological guidelines for the areas with significant restrictions for people who form part of the protected landscape. In this study, we understand the cost estimate of the social impact of national parks on the local population as a quantitative calculation of the losses due to restrictions on their ownership rights to land and property assets. The methodological approach consists in considering the category of losses as a sum total of the actual damage and lost profits. The assessment algorithm includes three stages: systematization of social impacts on citizens, development of indicators and data collection, and calculation of actual damage to the population and lost profits. The assessment is performed using the example of the Tunkinsky National Park located in the Tunkinsky municipal district of the Republic of Buryatia, a region of the Russian Federation, where there are 14 rural settlements with a population of more than 20,000 people. The results of the calculations show that the losses of the rural population due to legal restrictions on the registration of land dealings amount to 170.4 million USD. Taking into account the potential amount of administrative fines and the value of property subject to demolition, the losses amount to 239.2 million USD. It is more than an order of magnitude greater than the amount of own revenues of the Tunkinsky municipal district in 2011–2019. The results obtained demonstrate the real picture of the impact of restrictions on the rights of local people to land within the boundaries of national parks and are useful for developing measures to account for their interests and include protected areas in the socio-economic development of regions. The methodological approach developed by the authors can be used in other national parks, where it is necessary to optimize the policy of improving land use for local residents.
Correction
Charles A. Coomer, Irene Carlon-Andres, Maro Iliopoulou, Michael L. Dustin, Ewoud B. Compeer, Alex A. Compton, Sergi Padilla-Parra
PLOS Pathogens, Volume 17; doi:10.1371/journal.ppat.1009584

Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0244122

Abstract:
During last years “irreproducibility” became a general problem in omics data analysis due to the use of sophisticated and poorly described computational procedures. For avoiding misleading results, it is necessary to inspect and reproduce the entire data analysis as a unified product. Reproducible Research (RR) provides general guidelines for public access to the analytic data and related analysis code combined with natural language documentation, allowing third-parties to reproduce the findings. We developed easyreporting, a novel R/Bioconductor package, to facilitate the implementation of an RR layer inside reports/tools. We describe the main functionalities and illustrate the organization of an analysis report using a typical case study concerning the analysis of RNA-seq data. Then, we show how to use easyreporting in other projects to trace R functions automatically. This latter feature helps developers to implement procedures that automatically keep track of the analysis steps. Easyreporting can be useful in supporting the reproducibility of any data analysis project and shows great advantages for the implementation of R packages and GUIs. It turns out to be very helpful in bioinformatics, where the complexity of the analyses makes it extremely difficult to trace all the steps and parameters used in the study.
Hui Zhang, Tianhong Zhao, , Hang Wu, Yang Yang, Xuefa Wen
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0237684

Abstract:
Interannual variability (IAV) in net ecosystem carbon production (NEP) plays an important role in the processes of the carbon cycle, but the long-term trends in NEP and the climatic and biotic control of IAV in NEP still remain unclear in agroecosystems. We investigated interannual variability in NEP, expressed as annual values and anomalies, and its climatic and biotic controls using an eddy-covariance dataset for 2005–2018 for rain-fed spring maize in northeastern China. Average annual NEP was 270±31 g C m−2yr −1, with no significant changes over time. The effects on interannual variability in NEP of gross ecosystem productivity (GEP) that was mainly controlled by soil water content (SWC) and leaf area index (LAI), were more than those of respiration (RE) that was controlled by temperature and LAI. Further, maximum daily NEP (NEPmax) that was dominated by summer vapor pressure deficit explained the largest fraction of annual anomalies in NEP, followed by carbon dioxide uptake period (CUP) that was defined by the beginning date (BDOY) and the end date (EDOY) of CUP. The variability in BDOY was mainly determined by spring precipitation and the effective accumulated temperature, and the variability in EDOY was determined by autumn precipitation, SWC and LAI. NEP may decrease with declining precipitation in the future due to decreasing GEP, NEPmax, or CUP, and irrigation and residues cover may be useful in efforts to maintain current NEP levels. Our results indicate that interannual variability in NEP in agroecosystems may be more sensitive to changes in water conditions (such as precipitation, SWC and VPD) induced by climate changes, while temperature may be an important indirect factor when VPD is dominated.
Daniel Graeber, , Carsten Schröder
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0248372

Abstract:
Several vaccines against COVID-19 have now been developed and are already being rolled out around the world. The decision whether or not to get vaccinated has so far been left to the individual citizens. However, there are good reasons, both in theory as well as in practice, to believe that the willingness to get vaccinated might not be sufficiently high to achieve herd immunity. A policy of mandatory vaccination could ensure high levels of vaccination coverage, but its legitimacy is doubtful. We investigate the willingness to get vaccinated and the reasons for an acceptance (or rejection) of a policy of mandatory vaccination against COVID-19 in June and July 2020 in Germany based on a representative real time survey, a random sub-sample (SOEP-CoV) of the German Socio-Economic Panel (SOEP). Our results show that about 70 percent of adults in Germany would voluntarily get vaccinated against the coronavirus if a vaccine without side effects was available. About half of residents of Germany are in favor, and half against, a policy of mandatory vaccination. The approval rate for mandatory vaccination is significantly higher among those who would get vaccinated voluntarily (around 60 percent) than among those who would not get vaccinated voluntarily (27 percent). The individual willingness to get vaccinated and acceptance of a policy of mandatory vaccination correlates systematically with socio-demographic and psychological characteristics of the respondents. We conclude that as far as people’s declared intentions are concerned, herd immunity could be reached without a policy of mandatory vaccination, but that such a policy might be found acceptable too, were it to become necessary.
, Aleksandra Torbica, Chris P. Gale, Luigi Tavazzi, Panos Vardas,
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0250931

Abstract:
Background Although many studies have described patient-level risk factors for outcomes in heart failure (HF), health care structural determinants remain largely unexplored. This research reports patient-, hospital- and country-level characteristics associated with 1-year all-cause mortality among patients with chronic HF, and investigates geographic and hospital variation in mortality. Methods and findings We included 9,277 patients with chronic HF enrolled between May 2011 and November 2017 in the prospective cohort study European Society of Cardiology Heart Failure Long Term registry across 142 hospitals, located in 22 countries. Mean age of the selected outpatients was 65 years (sd 13.2) and 28% were female. The all-cause 1-year mortality rate per 100 person-years was 7.1 (95% confidence interval (CI) 6.6–7.7), and varied between countries (median 6.8, IQR 5.6–11.2) and hospitals (median 7.8, IQR 5.2–12.4). Mortality was associated with age (incidence rate ratio 1.03, 95% CI 1.02–1.04), diabetes mellitus (1.37, 1.15–1.63), peripheral artery disease (1.56, 1.27–1.92), New York Heart Association class III/IV (1.91, 1.60–2.30), treatment with angiotensin-converting enzyme inhibitor and angiotensin receptor antagonists (0.71, 0.57–0.87) and HF clinic (0.64, 0.46–0.89). No other hospital-level characteristics, and no country-level healthcare characteristics were associated with 1-year mortality, with case-mix standardised variance between countries being very low (1.83e-06) and higher for hospitals (0.372). Conclusions All-cause mortality at 1 year among outpatients with chronic HF varies between countries and hospitals, and is associated with patient characteristics and the availability of hospital HF clinics. After full adjustment for clinical, hospital and country variables, between-country variance was negligible while between-hospital variance was evident.
, , Simone Kunzelmann, Shahid Mehmood, , , Peter Cherepanov,
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251261

Abstract:
Chl1 is a member of the XPD family of 5’-3’ DNA helicases, which perform a variety of roles in genome maintenance and transmission. They possess a variety of unique structural features, including the presence of a highly variable, partially-ordered insertion in the helicase domain 1. Chl1 has been shown to be required for chromosome segregation in yeast due to its role in the formation of persistent chromosome cohesion during S-phase. Here we present structural and biochemical data to show that Chl1 has the same overall domain organisation as other members of the XPD family, but with some conformational alterations. We also present data suggesting the insert domain in Chl1 regulates its DNA binding.
Rachele M. Bochart, Kathleen Busman-Sahay, Stephen Bondoc, David W. Morrow, Alexandra M. Ortiz, Christine M. Fennessey, Miranda B. Fischer, Oriene Shiel, Tonya Swanson, Christine M. Shriver-Munsch, et al.
PLOS Pathogens, Volume 17; doi:10.1371/journal.ppat.1009565

Abstract:
Here, we assessed the efficacy of a short-course multimodal therapy (enrofloxacin, azithromycin, fenbendazole, and paromomycin) to eliminate common macaque endemic pathogens (EPs) and evaluated its impact on gastrointestinal (GI) microbiota, mucosal integrity, and local and systemic inflammation in sixteen clinically healthy macaques. Treatment combined with expanded practices resulted in successful maintenance of rhesus macaques (RM) free of common EPs, with no evidence of overt microbiota diversity loss or dysbiosis and instead resulted in a more defined luminal microbiota across study subjects. Creation of a GI pathogen free (GPF) status resulted in improved colonic mucosal barrier function (histologically, reduced colonic MPO+, and reduced pan-bacterial 16s rRNA in the MLN), reduced local and systemic innate and adaptive inflammation with reduction of colonic Mx1 and pSTAT1, decreased intermediate (CD14+CD16+) and non-classical monocytes (CD14-CD16+), reduced populations of peripheral dendritic cells, Ki-67+ and CD38+ CD4+ T cells, Ki-67+IgG+, and Ki-67+IgD+ B cells indicating lower levels of background inflammation in the distal descending colon, draining mesenteric lymph nodes, and systemically in peripheral blood, spleen, and axillary lymph nodes. A more controlled rate of viral acquisition resulted when untreated and treated macaques were challenged by low dose intrarectal SIVmac239x, with an ~100 fold increase in dose required to infect 50% (AID50) of the animals receiving treatment compared to untreated controls. Reduction in and increased consistency of number of transmitted founder variants resulting from challenge seen in the proof of concept study directly correlated with post-treatment GPF animal’s improved barrier function and reduction of key target cell populations (Ki-67+ CD4+T cells) at the site of viral acquisition in the follow up study. These data demonstrate that a therapeutic and operational strategy can successfully eliminate varying background levels of EPs and their associated aberrant immunomodulatory effects within a captive macaque cohort, leading to a more consistent, better defined and reproducible research model.
Christine Damrau, ,
PLOS Biology, Volume 19; doi:10.1371/journal.pbio.3001228

Abstract:
The biogenic amine octopamine (OA) and its precursor tyramine (TA) are involved in controlling a plethora of different physiological and behavioral processes. The tyramine-ß-hydroxylase (tßh) gene encodes the enzyme catalyzing the last synthesis step from TA to OA. Here, we report differential dominance (from recessive to overdominant) of the putative null tßh nM18 allele in 2 behavioral measures in Buridan’s paradigm (walking speed and stripe deviation) and in proboscis extension (sugar sensitivity) in the fruit fly Drosophila melanogaster. The behavioral analysis of transgenic tßh expression experiments in mutant and wild-type flies as well as of OA and TA receptor mutants revealed a complex interaction of both aminergic systems. Our analysis suggests that the different neuronal networks responsible for the 3 phenotypes show differential sensitivity to tßh gene expression levels. The evidence suggests that this sensitivity is brought about by a TA/OA opponent system modulating the involved neuronal circuits. This conclusion has important implications for standard transgenic techniques commonly used in functional genetics.
, Krystel Moussally, Chantal Lakis, Maya El-Hajj, Sola Bahous, Carla Peruzzo, Anthony Reid, Jeffrey K. Edwards
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251316

Abstract:
Introduction Low adherence to medications, specifically in patients with Diabetes (DM) and Hypertension (HTN), and more so in refugee settings, remains a major challenge to achieving optimum clinical control in these patients. We aimed at determining the self-reported medication adherence prevalence and its predictors and exploring reasons for low adherence among these patients. Methods A mixed-methods study was conducted at Médecins Sans Frontières non-communicable diseases primary care center in the Shatila refugee camp in Beirut, Lebanon in October 2018. Data were collected using the validated Arabic version of the 8-items Morisky Medication Adherence Scale (MMAS-8) concurrently followed by in-depth interviews to explore barriers to adherence in patients with DM and/or HTN. Predictors of adherence were separately assessed using logistic regression with SPSS© version 20. Manual thematic content analysis was used to analyze the qualitative data. Results Of the 361 patients included completing the MMAS, 70% (n = 251) were moderately to highly adherent (MMAS-8 score = 6 to 8), while 30% (n = 110) were low-adherent (MMAS-8 score<6). Patients with DM-1 were the most likely to be moderately to highly adherent (85%; n = 29). Logistic regression analysis showed that patients with a lower HbA1C were 75% more likely to be moderately to highly adherent [(OR = 0.75 (95%CI 0.63–0.89), p-value 0.001]. Factors influencing self-reported moderate and high adherence were related to the burden of the disease and its treatment, specifically insulin, the self-perception of the disease outcomes and the level of patient’s knowledge about the disease and other factors like supportive family and healthcare team. Conclusion Adherence to DM and HTN was good, likely due to a patient-centered approach along with educational interventions. Future studies identifying additional factors and means addressing the barriers to adherence specific to the refugee population are needed to allow reaching optimal levels of adherence and design well-informed intervention programs.
, Emily E. Franchett, Clariana V. Ramos de Oliveira, Karima Rehmani,
PLOS Medicine, Volume 18; doi:10.1371/journal.pmed.1003602

Abstract:
Background Parents are the primary caregivers of young children. Responsive parent–child relationships and parental support for learning during the earliest years of life are crucial for promoting early child development (ECD). We conducted a global systematic review and meta-analysis to evaluate the effectiveness of parenting interventions on ECD and parenting outcomes. Methods and findings We searched MEDLINE, Embase, PsycINFO, CINAHL, Web of Science, and Global Health Library for peer-reviewed, published articles from database inception until November 15, 2020. We included randomized controlled trials (RCTs) of parenting interventions delivered during the first 3 years of life that evaluated at least 1 ECD outcome. At least 2 reviewers independently screened, extracted data, and assessed study quality from eligible studies. ECD outcomes included cognitive, language, motor, and socioemotional development, behavior problems, and attachment. Parenting outcomes included parenting knowledge, parenting practices, parent–child interactions, and parental depressive symptoms. We calculated intervention effect sizes as the standardized mean difference (SMD) and estimated pooled effect sizes for each outcome separately using robust variance estimation meta-analytic approaches. We used random-effects meta-regression models to assess potential effect modification by country-income level, child age, intervention content, duration, delivery, setting, and study quality. This review was registered with PROSPERO (CRD42018092458 and CRD42018092461). Of the 11,920 articles identified, we included 111 articles representing 102 unique RCTs. Pooled effect sizes indicated positive benefits of parenting interventions on child cognitive development (SMD = 0.32, 95% CI [confidence interval]: 0.23, 0.40, P < 0.001), language development (SMD = 0.28, 95% CI: 0.18 to 0.37, P < 0.001), motor development (SMD = 0.24, 95% CI: 0.15 to 0.32, P < 0.001), socioemotional development (SMD = 0.19, 95% CI: 0.10 to 0.28, P < 0.001), and attachment (SMD = 0.29, 95% CI: 0.18 to 0.40, P < 0.001) and reductions in behavior problems (SMD = −0.13, 95% CI: −0.18 to −0.08, P < 0.001). Positive benefits were also found on parenting knowledge (SMD = 0.56, 95% CI: 0.33 to 0.79, P < 0.001), parenting practices (SMD = 0.33, 95% CI: 0.22 to 0.44, P < 0.001), and parent–child interactions (SMD = 0.39, 95% CI: 0.24 to 0.53, P < 0.001). However, there was no significant reduction in parental depressive symptoms (SMD = −0.07, 95% CI: −0.16 to 0.02, P = 0.08). Subgroup analyses revealed significantly greater effects on child cognitive, language, and motor development, and parenting practices in low- and middle-income countries compared to high-income countries; and significantly greater effects on child cognitive development, parenting knowledge, parenting practices, and parent–child interactions for programs that focused on responsive caregiving compared to those that did not. On the other hand, there was no clear evidence of effect modification by child age, intervention duration, delivery, setting, or study risk of bias. Study limitations include considerable unexplained heterogeneity, inadequate reporting of intervention content and implementation, and varying quality of evidence in terms of the conduct of trials and robustness of outcome measures used across studies. Conclusions Parenting interventions for children during the first 3 years of life are effective for improving ECD outcomes and enhancing parenting outcomes across low-, middle-, and high-income countries. Increasing implementation of effective and high-quality parenting interventions is needed globally and at scale in order to support parents and enable young children to achieve their full developmental potential.
, Ellen Schulz-Kornas, Ottmar Kullmer, Marcel M. Janocha, Roman M. Wittig,
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251309

Abstract:
Dental wear analyses have been widely used to interpret the dietary ecology in primates. However, it remains unclear to what extent a combination of wear analyses acting at distinct temporal scales can be beneficial in interpreting the tooth use of primates with a high variation in their intraspecific dietary ecology. Here, we combine macroscopic tooth wear (occlusal fingerprint analysis, long-term signals) with microscopic 3D surface textures (short-term signals) exploring the tooth use of a historical western chimpanzee population from northeastern Liberia with no detailed dietary records. We compare our results to previously published tooth wear and feeding data of the extant and continually monitored chimpanzees of Taї National Park in Ivory Coast. Macroscopic tooth wear results from molar wear facets of the Liberian population indicate only slightly less wear when compared to the Taї population. This suggests similar long-term feeding behavior between both populations. In contrast, 3D surface texture results show that Liberian chimpanzees have many and small microscopic wear facet features that group them with those Taї chimpanzees that knowingly died during dry periods. This coincides with historical accounts, which indicate that local tribes poached and butchered the Liberian specimens during dust-rich dry periods. In addition, Liberian females and males differ somewhat in their 3D surface textures, with females having more microscopic peaks, smaller hill and dale areas and slightly rougher wear facet surfaces than males. This suggests a higher consumption of insects in Liberian females compared to males, based on similar 3D surface texture patterns previously reported for Taї chimpanzees. Our study opens new options for uncovering details of feeding behaviors of chimpanzees and other living and fossil primates, with macroscopic tooth wear tracing the long-term dietary and environmental history of a single population and microscopic tooth wear addressing short-term changes (e.g. seasonality).
, Wei Lin, Chia Chieh Goh, Feng Gao, Nicholas Larson, Joseph Wahl, Thomas V. Bilfinger, Arjun G. Yodh, Thomas F. Floyd
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251271

Abstract:
Spinal cord ischemia leads to iatrogenic injury in multiple surgical fields, and the ability to immediately identify onset and anatomic origin of ischemia is critical to its management. Current clinical monitoring, however, does not directly measure spinal cord blood flow, resulting in poor sensitivity/specificity, delayed alerts, and delayed intervention. We have developed an epidural device employing diffuse correlation spectroscopy (DCS) to monitor spinal cord ischemia continuously at multiple positions. We investigate the ability of this device to localize spinal cord ischemia in a porcine model and validate DCS versus Laser Doppler Flowmetry (LDF). Specifically, we demonstrate continuous (>0.1Hz) spatially resolved (3 locations) monitoring of spinal cord blood flow in a purely ischemic model with an epidural DCS probe. Changes in blood flow measured by DCS and LDF were highly correlated (r = 0.83). Spinal cord blood flow measured by DCS caudal to aortic occlusion decreased 62%. This monitor demonstrated a sensitivity of 0.87 and specificity of 0.91 for detection of a 25% decrease in flow. This technology may enable early identification and critically important localization of spinal cord ischemia.
, , Robert J. Scott, Benjamin Kerr
PLOS Pathogens, Volume 17; doi:10.1371/journal.ppat.1009528

Abstract:
Tradeoff theory, which postulates that virulence provides both transmission costs and benefits for pathogens, has become widely adopted by the scientific community. Although theoretical literature exploring virulence-tradeoffs is vast, empirical studies validating various assumptions still remain sparse. In particular, truncation of transmission duration as a cost of virulence has been difficult to quantify with robust controlled in vivo studies. We sought to fill this knowledge gap by investigating how transmission rate and duration were associated with virulence for infectious hematopoietic necrosis virus (IHNV) in rainbow trout (Oncorhynchus mykiss). Using host mortality to quantify virulence and viral shedding to quantify transmission, we found that IHNV did not conform to classical tradeoff theory. More virulent genotypes of the virus were found to have longer transmission durations due to lower recovery rates of infected hosts, but the relationship was not saturating as assumed by tradeoff theory. Furthermore, the impact of host mortality on limiting transmission duration was minimal and greatly outweighed by recovery. Transmission rate differences between high and low virulence genotypes were also small and inconsistent. Ultimately, more virulent genotypes were found to have the overall fitness advantage, and there was no apparent constraint on the evolution of increased virulence for IHNV. However, using a mathematical model parameterized with experimental data, it was found that host culling resurrected the virulence tradeoff and provided low virulence genotypes with the advantage. Human-induced or natural culling, as well as host population fragmentation, may be some of the mechanisms by which virulence diversity is maintained in nature. This work highlights the importance of considering non-classical virulence tradeoffs.
Lucy D. Guarnieri, Sara E. McBride, Eleanor Groden,
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251497

Abstract:
The blacklegged tick (Ixodes scapularis) and the invasive European fire ant (Myrmica rubra) are both expanding throughout their sympatric range in coastal New England. Ixodes scapularis is the primary vector of the bacterium Borrelia burgdorferi, which is the causative agent of Lyme disease, and Mount Desert Island, Maine, home to Acadia National Park, currently is affected by a high Lyme disease burden. Ticks have many natural predators, including ants, although no previous studies have investigated interactions between these two species. To test the hypothesis that the presence of M. rubra alters I. scapularis abundance, we collected ticks by drag-sampling at eight ant-infested sites and eight uninfested control sites in Acadia National Park. We found that nymph density was significantly higher at ant-infested sites, while larval density was significantly higher at control sites. In addition, we conducted a laboratory bioassay to measure M. rubra aggression against I. scapularis larvae, nymphs, and adults and Dermacentor variabilis adults, and found that ant aggression was significantly higher against D. variabilis adults than I. scapularis adults. Our findings support the hypothesis that M. rubra has divergent effects across I. scapularis life stages, and we discuss possible ecological mechanisms, including optimal microclimate and predation, that could promote density of nymphs while inhibiting density of larvae.
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0240706

Abstract:
Hibernating mammals exhibit unique metabolic and physiological phenotypes that have potential applications in medicine or spaceflight, yet our understanding of the genetic basis and molecular mechanisms of hibernation is limited. The meadow jumping mouse, a small North American hibernator, exhibits traits–including a short generation time–that would facilitate genetic approaches to hibernation research. Here we report the collection, captive breeding, and laboratory hibernation of meadow jumping mice. Captive breeders in our colony produced a statistically significant excess of male offspring and a large number of all-male and all-female litters. We confirmed that short photoperiod induced pre-hibernation fattening, and cold ambient temperature facilitated entry into hibernation. During pre-hibernation fattening, food consumption exhibited non-linear dependence on both body mass and temperature, such that food consumption was greatest in the heaviest animals at the coldest temperatures. Meadow jumping mice exhibited a strong circadian rhythm of nightly activity that was disrupted during the hibernation interval. We conclude that it is possible to study hibernation phenotypes using captive-bred meadow jumping mice in a laboratory setting.
Kumiko Okino, Mitsuhiro Aoki, Masahiro Yamane, Chikashi Kohmura
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251532

Abstract:
Objective The purpose of this study is to investigate the physical changes of the lower leg muscles in the compartment by observing the changes in the shear wave velocity of the gastrocnemius, soleus and tibialis anterior muscles with time in the sitting position for 2 hours and after elevation of the lower leg. Materials and methods The subjects were 24 healthy adult males (average age 26.6 years). Shear wave velocity was measured by Aplio 500 in immobilized leg immediately after the start of sitting, 60 minutes and 120 minutes after the start of sitting. After 120 minutes the subjects raised the lower leg for 3 minutes, then measured again. Results In the lateral and medial gastrocnemius, there was a significant increase in the velocity at 60 (1.58 ± 0.06, 1.70 ± 0.09 m/s) and 120 minutes (1.70 ± 0.10, 1.83 ± 0.11 m/s) after the start of the test (1.52 ± 0.06, 1.66 ± 0.10 m/s), respectively (p<0.01). In the soleus and the tibialis anterior, there was a significant increase in the velocity at 120 minutes (1.89 ± 0.17, 2.30 ± 0.24 m/s) compared to after the start (1.60 ± 0.15, 2.15 ± 0.26 m/s), respectively (p<0.01). In all muscles, there was a significant decrease in the velocity after the raising compared to that of 120 minutes (p<0.01). Conclusions It has been reported that the change of shear wave velocity with time is proportional to the intramuscular pressure in the leg compartment, and it is assumed that the increase of shear wave velocity in the 2-hour seated leg is due to fluid retention in extra-cellular space of the compartment.
Aminah M. I. Al-Awadi, Abdulaziz Isa AlJawder, Alyaa Mousa, Safa Taha,
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0248455

Abstract:
The Immune System-Released Activating Agent (ISRAA) was discovered as a novel molecule that functions as a mediator between the nervous and immune systems in response to a nervous stimulus following an immune challenge. This research investigated the role of ISRAA) in promoting the ontogeny of the mouse brain astrocytes. Astrocyte cultures were prepared from two-month-old BALB/c mice. Recombinant ISRAA protein was used to stimulate astrocyte cultures. Immunohistochemistry and ELISA were utilized to measure ISRAA and IFN-γ levels, IFN-γR expression and STAT1 nuclear translocation. MTT-assay was used to evaluate cellular survival and proliferation. To assess astrocyte cell lysates and tyrosine-phosphorylated proteins, SDS-PAGE and western blot were used. ISRAA was highly expressed in mouse embryonic astrocytes, depending on cell age. Astrocytes aged seven days (E7) showed increased proliferation and diminished differentiation, while 21-day-old (E21) astrocytes depicted reversed effects. IFN-γ was involved in the ISRAA action as ISRAA induced IFN-γ in both age groups, but only E21 astrocytes expressed IFN-γR. ISRAA stimulation of E21 resulted in tyrosine phosphorylation of numerous cellular proteins and the nuclear translocation of STAT1, a signalling pathway utilized by IFN-γ. The results suggest that ISRAA is involved in mouse brain development through the cytokine network involving IFN-γ.
PLOS Biology, Volume 19; doi:10.1371/journal.pbio.3001253

Abstract:
Sex/gender differences in the human brain attract attention far beyond the neuroscience community. Given the interest of nonspecialists, it is important that researchers studying human female–male brain difference assume greater responsibility for the accurate communication of their findings.
Daoxiu Xu, Xue Bai, Jing Xu, , Zijian Dong, Wenjie Shi, Fengyan Xu, Yanfeng Li, Mingyuan Liu,
PLOS Neglected Tropical Diseases, Volume 15; doi:10.1371/journal.pntd.0009408

Abstract:
Trichinellosis is a major foodborne parasitosis caused by Trichinella spiralis. In the present study, a serine protease gene from an adult T. spiralis (Ts-Adsp) cDNA library was cloned, expressed in Escherichia coli and purified by Ni-affinity chromatography. Previous studies of our laboratory have found that mice vaccinated with recombinant Ts-Adsp protein (rTs-Adsp) exhibited partial protection against T. spiralis infection. In this study, the protective effect of rTs-Adsp against T. spiralis infection in pigs was further explored. The cell-mediated and humoral immune responses induced by rTs-Adsp were measured, including the dynamic trends of specific antibody levels (IgG, IgG1, IgG2a and IgM), as well as the levels of cytokines (IFN-γ, IL-2, IL-4, and IL-10) in the serum. Moreover, the changes in T lymphocytes, B lymphocytes, and neutrophils were measured to evaluate cellular immune responses in pigs vaccinated with rTs-Adsp. The results indicated that a Th1-Th2 mixed immune response with Th1 predominant was induced by rTs-Adsp after vaccination. Flow cytometric analysis showed that the proportions of CD4+ T cells, B cells, and neutrophils in the immunized groups were significantly increased. Furthermore, pigs vaccinated with rTs-Adsp exhibited a 50.9% reduction in the muscle larvae burden, compare with pigs from the PBS group five weeks after challenged. Our results suggested that rTs-Adsp elicited partial protection and it could be a potential target molecule for preventing and controlling Trichinella transmission from pigs to human.
, Seifu Abera, Guiomar Basualdo, Roxanne P. Kerani, Farah Mohamed, Rahel Schwartz, Beyene Gebreselassie, Ahmed Ali,
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0250800

Abstract:
Studies of African immigrant health in the U.S. have traditionally focused on infectious diseases. However, the rising burden of non-communicable diseases (NCDs) indicates the increasing importance of general preventive health care. As part of a series of community health events designed for African-born individuals in King County, Washington, we administered key informant interviews (KIIs) with 16 health event participants, medical professionals, and community leaders to identify barriers and facilitators to use of preventive health care among African-born individuals. We used descriptive thematic analysis to organize barriers according to the socio-ecological model. Within the individual domain, KII participants identified lack of knowledge and awareness of preventive health benefits as barriers to engagement in care. Within the interpersonal domain, language and cultural differences frequently complicated relationships with health care providers. Within the societal and policy domains, healthcare costs, lack of insurance, and structural racism were also reported as major barriers. Participants identified community outreach with culturally competent and respectful providers as key elements of interventions to improve uptake. In conclusion, African immigrant communities face several barriers, ranging from individual to policy levels, to accessing health services, resulting in substantial unmet need for chronic disease prevention and treatment. Community-centered and -led care may help facilitate uptake and engagement in care.
Amanda M. Westman, , George J. Christ, Silvia S. Blemker
PLOS Computational Biology, Volume 17; doi:10.1371/journal.pcbi.1008937

Abstract:
Skeletal muscle possesses a remarkable capacity for repair and regeneration following a variety of injuries. When successful, this highly orchestrated regenerative process requires the contribution of several muscle resident cell populations including satellite stem cells (SSCs), fibroblasts, macrophages and vascular cells. However, volumetric muscle loss injuries (VML) involve simultaneous destruction of multiple tissue components (e.g., as a result of battlefield injuries or vehicular accidents) and are so extensive that they exceed the intrinsic capability for scarless wound healing and result in permanent cosmetic and functional deficits. In this scenario, the regenerative process fails and is dominated by an unproductive inflammatory response and accompanying fibrosis. The failure of current regenerative therapeutics to completely restore functional muscle tissue is not surprising considering the incomplete understanding of the cellular mechanisms that drive the regeneration response in the setting of VML injury. To begin to address this profound knowledge gap, we developed an agent-based model to predict the tissue remodeling response following surgical creation of a VML injury. Once the model was able to recapitulate key aspects of the tissue remodeling response in the absence of repair, we validated the model by simulating the tissue remodeling response to VML injury following implantation of either a decellularized extracellular matrix scaffold or a minced muscle graft. The model suggested that the SSC microenvironment and absence of pro-differentiation SSC signals were the most important aspects of failed muscle regeneration in VML injuries. The major implication of this work is that agent-based models may provide a much-needed predictive tool to optimize the design of new therapies, and thereby, accelerate the clinical translation of regenerative therapeutics for VML injuries.
, Jessica Kolic, Wen Wu, Kade Paterson
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251175

Abstract:
The development of walking in young toddlers is an important motor milestone. Walking patterns can differ widely amongst toddlers, and are characterised by unique biomechanical strategies. This makes comparisons between newly walking toddler’s and older children’s walking difficult. Little is currently understood regarding the effects of footwear on the gait in newly walking toddlers. A quasi-experimental pre-post study design was used to assess whether spatiotemporal parameters of gait, and in-shoe foot and lower limb kinematics, differed when walking barefoot and in soft-soled footwear in newly walking toddlers. There were 18 toddlers recruited, with 14 undergoing testing. The GAITRite system collected spatial and temporal data. The Vicon camera system collected kinematic data. The testing conditions included barefoot and footwear. Footwear tested was a commercially available soft soled shoe (Bobux XPLORER). Data was extracted directly from the GAITRite system and analysed. Walking in footwear did not change spatial or temporal data, however there were small but significant decreases in hip adduction/abduction range of motion (mean difference (MD) = 1.79°, 95% CI = -3.51 to -0.07, p = 0.04), knee flexion (MD = -7.63°, 95% CI = 2.70 to 12.55, p = 0.01), and knee flexion/extension range of movement (MD = 6.25°, 95% CI = -10.49 to -2.01, p = 0.01), and an increase in subtalar joint eversion (MD = 2.85°, 95% CI = 5.29 to -0.41, p = 0.03). Effect sizes were small for hip and ankle range, peak knee extension, and subtalar joint ranges (d<0.49), medium for knee flexion/extension range (d = 0.75) and large for peak knee flexion (d = 0.87). The magnitude of kinematic changes with soft-soled footwear were small thus the clinical importance of these findings is uncertain. Future longitudinal studies are needed to develop recommendations regarding footwear for newly walking toddlers.
, Hakaru Tamukoh
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0251329

Abstract:
In this study, we introduced a mixed-precision weights network (MPWN), which is a quantization neural network that jointly utilizes three different weight spaces: binary {−1,1}, ternary {−1,0,1}, and 32-bit floating-point. We further developed the MPWN from both software and hardware aspects. From the software aspect, we evaluated the MPWN on the Fashion-MNIST and CIFAR10 datasets. We systematized the accuracy sparsity bit score, which is a linear combination of accuracy, sparsity, and number of bits. This score allows Bayesian optimization to be used efficiently to search for MPWN weight space combinations. From the hardware aspect, we proposed XOR signed-bits to explore floating-point and binary weight spaces in the MPWN. XOR signed-bits is an efficient implementation equivalent to multiplication of floating-point and binary weight spaces. Using the concept from XOR signed bits, we also provide a ternary bitwise operation that is an efficient implementation equivalent to the multiplication of floating-point and ternary weight space. To demonstrate the compatibility of the MPWN with hardware implementation, we synthesized and implemented the MPWN in a field-programmable gate array using high-level synthesis. Our proposed MPWN implementation utilized up to 1.68-4.89 times less hardware resources depending on the type of resources than a conventional 32-bit floating-point model. In addition, our implementation reduced the latency up to 31.55 times compared to 32-bit floating-point model without optimizations.
, Philipp Gerlach, , Matthias Bogyi, Folke Brinkmann,
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0250387

Abstract:
Background Majority of active tuberculosis (TB) cases in children in low-incidence countries are due to rapid progression of infection (latent TB infection (LTBI)) to disease. We aimed to assess common practice for managing paediatric LTBI in Austria, Germany and Switzerland prior to the publication of the first joint national guideline for paediatric TB in 2017. Methods Online-based survey amongst pediatricians, practitioners and staff working in the public health sector between July and November 2017. Data analysis was conducted using IBM SPSS. Results A total of 191 individuals participated in the survey with 173 questionnaires included for final analysis. Twelve percent of respondents were from Austria, 60% from Germany and 28% from Switzerland. Proportion of children with LTBI and migrant background was estimated by the respondents to be >50% by 58%. Tuberculin skin test (TST) and interferon-γ-release-assay (IGRA), particularly Quantiferon-gold-test, were reported to be used in 86% and 88%, respectively. In children > 5 years with a positive TST or IGRA a chest x-ray was commonly reported to be performed (28%). Fifty-three percent reported to take a different diagnostic approach in children ≤ 5 years, mainly combining TST, IGRA and chest x-ray for initial testing (31%). Sixty-eight percent reported to prescribe isoniazid-monotherapy: for 9 (62%), or 6 months (6%), 31% reported to prescribe combination therapy of isoniazid and rifampicin. Dosing of isoniazid and rifampicin below current recommendations was reported by up to 22% of respondents. Blood-sampling before/during LTBI treatment was reported in >90% of respondents, performing a chest-X-ray at the end of treatment by 51%. Conclusion This survey showed reported heterogeneity in the management of paediatric LTBI. Thus, regular and easily accessible educational activities and national up-to-date guidelines are key to ensure awareness and quality of care for children and adolescents with LTBI in low-incidence countries.
Nak Won Rim, , Coltan Scrivner,
Published: 10 May 2021
PLOS ONE, Volume 16; doi:10.1371/journal.pone.0250170

Abstract:
Many eye-tracking data analyses rely on the Area-of-Interest (AOI) methodology, which utilizes AOIs to analyze metrics such as fixations. However, AOI-based methods have some inherent limitations including variability and subjectivity in shape, size, and location of AOIs. In this article, we propose an alternative approach to the traditional AOI dwell time analysis: Weighted Sum Durations (WSD). This approach decreases the subjectivity of AOI definitions by using Points-of-Interest (POI) while maintaining interpretability. In WSD, the durations of fixations toward each POI is weighted by the distance from the POI and summed together to generate a metric comparable to AOI dwell time. To validate WSD, we reanalyzed data from a previously published eye-tracking study (n = 90). The re-analysis replicated the original findings that people gaze less towards faces and more toward points of contact when viewing violent social interactions.
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