How to normalize metatranscriptomic count data for differential expression analysis
Background Differential expression analysis on the basis of RNA-Seq count data has become a standard tool in transcriptomics. Several studies have shown that prior normalization of the data is crucial for a reliable detection of transcriptional differences. Until now it has not been clear whether and how the transcriptomic approach can be used for differential expression analysis in metatranscriptomics. Methods We propose a model for differential expression in metatranscriptomics that explicitly accounts for variations in the taxonomic composition of transcripts across different samples. As a main consequence the correct normalization of metatranscriptomic count data under this model requires the taxonomic separation of the data into organism-specific bins. Then the taxon-specific scaling of organism profiles yields a valid normalization and allows us to recombine the scaled profiles into a metatranscriptomic count matrix. This matrix can then be analyzed with statistical tools for transcriptomic count data. For taxon-specific scaling and recombination of scaled counts we provide a simple R script. Results When applying transcriptomic tools for differential expression analysis directly to metatranscriptomic data with an organism-independent (global) scaling of counts the resulting differences may be difficult to interpret. The differences may correspond to changing functional profiles of the contributing organisms but may also result from a variation of taxonomic abundances. Taxon-specific scaling eliminates this variation and therefore the resulting differences actually reflect a different behavior of organisms under changing conditions. In simulation studies we show that the divergence between results from global and taxon-specific scaling can be drastic. In particular, the variation of organism abundances can imply a considerable increase of significant differences with global scaling. Also, on real metatranscriptomic data, the predictions from taxon-specific and global scaling can differ widely. Our studies indicate that in real data applications performed with global scaling it might be impossible to distinguish between differential expression in terms of transcriptomic changes and differential composition in terms of changing taxonomic proportions. Conclusions As in transcriptomics, a proper normalization of count data is also essential for differential expression analysis in metatranscriptomics. Our model implies a taxon-specific scaling of counts for normalization of the data. The application of taxon-specific scaling consequently removes taxonomic composition variations from functional profiles and therefore provides a clear interpretation of the observed functional differences.
Klingenberg et al. (2017) How to normalize metatranscriptomic count data for differential expression analysis. PeerJ 5:e3859 https://doi.org/10.7717/peerj.3859
Exploring the psychological health of emergency dispatch centre operatives: a systematic review and narrative synthesis
Background The study objective was to investigate and synthesize available evidence relating to the psychological health of Emergency Dispatch Centre (EDC) operatives, and to identify key stressors experienced by EDC operatives. Methods Eight electronic databases (Embase, PubMed, Medline, CINAHL, PsycInfo, PsycArticles, The Psychology and Behavioural Sciences Collection, and Google Scholar) were searched. All study designs were included, and no date limits were set. Studies were included if they were published in English, and explored the psychological health of any EDC operatives, across fire, police, and emergency medical services. Studies were excluded if they related solely to other emergency workers, such as police officers or paramedics. Methodological quality of included studies was assessed using checklists adapted from the Critical Appraisal Skills Programme. A narrative synthesis was conducted, using thematic analysis. Results A total of 16 articles were included in the review. Two overarching themes were identified during the narrative synthesis: ‘Organisational and Operational Factors’ and ‘Interactions with Others’. Stressors identified included being exposed to traumatic calls, lacking control over high workload, and working in under-resourced and pressured environments. Lack of support from management and providing an emotionally demanding service were additional sources of stress. Peer support and social support from friends and family were helpful in managing work-related stress. Discussion EDC operatives experience stress as a result of their work, which appears to be related to negative psychological health outcomes. Future research should explore the long-term effects of this stress, and the potential for workplace interventions to alleviate the negative impacts on psychological health. PROSPERO Registration Number CRD42014010806.
Golding et al. (2017) Exploring the psychological health of emergency dispatch centre operatives: a systematic review and narrative synthesis. PeerJ 5:e3735 https://doi.org/10.7717/peerj.3735
Microbial community diversity patterns are related to physical and chemical differences among temperate lakes near Beaver Island, MI
Lakes are dynamic and complex ecosystems that can be influenced by physical, chemical, and biological processes. Additionally, individual lakes are often chemically and physically distinct, even within the same geographic region. Here we show that differences in physicochemical conditions among freshwater lakes located on (and around) the same island, as well as within the water column of each lake, are significantly related to aquatic microbial community diversity. Water samples were collected over time from the surface and bottom-water within four freshwater lakes located around Beaver Island, MI within the Laurentian Great Lakes region. Three of the sampled lakes experienced seasonal lake mixing events, impacting either O2, pH, temperature, or a combination of the three. Microbial community alpha and beta diversity were assessed and individual microbial taxa were identified via high-throughput sequencing of the 16S rRNA gene. Results demonstrated that physical and chemical variability (temperature, dissolved oxygen, and pH) were significantly related to divergence in the beta diversity of surface and bottom-water microbial communities. Despite its correlation to microbial community structure in unconstrained analyses, constrained analyses demonstrated that dissolved organic carbon (DOC) concentration was not strongly related to microbial community structure among or within lakes. Additionally, several taxa were correlated (either positively or negatively) to environmental variables, which could be related to aerobic and anaerobic metabolisms. This study highlights the measurable relationships between environmental conditions and microbial communities within freshwater temperate lakes around the same island.
Hengy et al. (2017) Microbial community diversity patterns are related to physical and chemical differences among temperate lakes near Beaver Island, MI. PeerJ 5:e3937 https://doi.org/10.7717/peerj.3937
Tolerance: the forgotten child of plant resistance
Plant resistance against insect herbivory has greatly focused on antibiosis, whereby the plant has a deleterious effect on the herbivore, and antixenosis, whereby the plant is able to direct the herbivore away from it. Although these two types of resistance may reduce injury and yield loss, they can produce selection pressures on insect herbivores that lead to pest resistance. Tolerance, on the other hand, is a more sustainable pest management strategy because it involves only a plant response and therefore does not cause evolution of resistance in target pest populations. Despite its attractive attributes, tolerance has been poorly studied and understood. In this critical, interpretive review, we discuss tolerance to insect herbivory and the biological and socioeconomic factors that have limited its use in plant resistance and integrated pest management. First, tolerance is difficult to identify, and the mechanisms conferring it are poorly understood. Second, the genetics of tolerance are mostly unknown. Third, several obstacles hinder the establishment of high-throughput phenotyping methods for large-scale screening of tolerance. Fourth, tolerance has received little attention from entomologists because, for most, their primary interest, research training, and funding opportunities are in mechanisms which affect pest biology, not plant biology. Fifth, the efforts of plant resistance are directed at controlling pest populations rather than managing plant stress. We conclude this paper by discussing future research and development activities.
Peterson et al. (2017) Tolerance: the forgotten child of plant resistance. PeerJ 5:e3934 https://doi.org/10.7717/peerj.3934
L1 and L2 reading skills in Dutch adolescents with a familial risk of dyslexia
Background The present study investigated differences in reading and spelling outcomes in Dutch and English as a second language (ESL) in adolescents with a high familial risk of dyslexia, of whom some have developed dyslexia (HRDys) while others have not (HRnonDys), in comparison to a low familial risk control group without dyslexia (LRnonDys). This allowed us to investigate the persistence of dyslexia in the first language (L1) and the effect of dyslexia on the second language (L2), which has, in this case, a lower orthographic transparency. Furthermore, the inclusion of the HRnonDys group allowed us to investigate the continuity of the familial risk of dyslexia, as previous studies observed that the HRnonDys group often scores in between the HRDys and LRnonDys group, and whether these readers without reading deficits in Dutch, have more reading difficulties in ESL. Methods The data of three groups of adolescents were analyzed; 27 LRnonDys, 25 HRdys 25 HRnonDys. The mean age was 14;1 years; months, and 37 were male. All were native speakers of Dutch, attended regular secondary education (grade 7–10), and were non-native speakers of English. Using MANOVA the groups were compared on Dutch and English word reading fluency (WRF), spelling and vocabulary, Dutch pseudoword and loanword reading fluency, phonological awareness (PA), rapid automatized naming (RAN), and verbal short term and working memory. A repeated measures ANOVA was used to compare English and Dutch WRF, spelling and vocabulary directly within the three groups. Results The analyses revealed that the HRDys group had a deficit in both reading and spelling in Dutch and ESL. They also performed poorer than the LRnonDys group on all other measures. Effect sizes were especially large for pseudoword reading and the reaction times during the PA task. The HRnonDys group scored generally poorer than the LRnonDys group but this difference was only significant for Dutch pseudoword reading, PA reaction times and verbal short term memory. In general the HRDys and HRnonDys group scored similar in Dutch and English, except for English WRF where the HRDys group scored slightly better than expected based on their Dutch WRF. Discussion There was a high persistence of dyslexia. Adolescents with dyslexia had large impairments in reading and spelling, and reading related measures, both in Dutch and ESL. Despite high inter-individual differences, an overall three-step pattern was observed. Adolescents in the HRnonDys group scored in between the HRDys and LRnonDys group, supporting the polygenetic origin of dyslexia and the continuity of the familial risk of dyslexia. The lower orthographic transparency did not have a negative effect on L2 reading, spelling and vocabulary, both in the HRnonDys and HRDys group. The latter group performed slightly better than expected in L2, which may be a result of the massive exposure to English and high motivation to use English by adolescents.
van Setten et al. (2017) L1 and L2 reading skills in Dutch adolescents with a familial risk of dyslexia. PeerJ 5:e3895 https://doi.org/10.7717/peerj.3895
Investigating the level of agreement of two positioning protocols when using dual energy X-ray absorptiometry in the assessment of body composition
Background Dual energy X-ray absorptiometry (DXA) is a commonly used instrument for analysing segmental body composition (BC). The information from the scan guides the clinician in the treatment of conditions such as obesity and can be used to monitor recovery of lean mass following injury. Two commonly used DXA positioning protocols have been identified—the Nana positioning protocol and the National Health and Nutrition Examination Survey (NHANES). Both protocols have been shown to be reliable. However, only one study has assessed the level of agreement between the protocols and ascertained the participants’ preference of protocol based upon comfort. Given the paucity of research in the field and the growing use of DXA in both healthy and pathological populations further research determining the most appropriate positioning protocol is warranted. Therefore, the aims of this study were to assess the level of agreement between results from the NHANES protocol and Nana protocol, and the participants’ preference of protocol based on comfort. Methods Thirty healthy participants (15 males, 15 females, aged 23–59 years) volunteered to participate in this study. These participants underwent two whole body DXA scans in a single morning (Nana positioning protocol and NHANES positioning protocol), in a randomised order. Each participant attended for scanning wearing minimal clothing and having fasted overnight, refrained from exercise in the past 24 h and voided their bladders. Level of agreement, comparing NAHNES to Nana protocol was assessed using an intra-class correlation coefficient (ICC), concordance correlation coefficient (CCC) and percentage change in mean. Limit of agreement comparing the two protocols were assessed using plots, mean difference and confidence limits. Participants were asked to indicate the protocol they found most comfortable. Results When assessing level of agreement between protocols both the ICC and CCC scores were very high and ranged from 0.987 to 0.997 for whole body composition, indicating excellent agreement between the Nana and NHANES protocols. Regional analysis (arms, legs, trunk) ICC scores, ranged between 0.966 and 0.996, CCC ranged between 0.964 and 0.997, change in mean percentage ranged between −0.58% and 0.37% which indicated a very high level of agreement. Limit of agreement analysis using mean difference ranged between −0.223 and 0.686 kg and 95% CL produced results ranging between −1.262 kg and 1.630 kg. The majority (80%) of participants found the NHANES positioning protocol more comfortable. Discussion This study reveals a strong level of agreement as illustrated by high ICC’s and CCC’s between the positioning protocols, however systematic bias within limit of agreement plot and a large difference in 95% confidence limits indicates that the protocols should not be interchanged when assessing an individual. The NHANES protocol affords greater participant comfort.
Shiel et al. (2017) Investigating the level of agreement of two positioning protocols when using dual energy X-ray absorptiometry in the assessment of body composition. PeerJ 5:e3880 https://doi.org/10.7717/peerj.3880
Tumor-infiltrating CD8+ lymphocytes predict different clinical outcomes in organ- and non-organ-confined urothelial carcinoma of the bladder following radical cystectomy
Tumor-infiltrating lymphocytes (TILs) are associated with better clinical outcomes in many tumors. TILs represent a cell-mediated immune response against the carcinoma. CD8+ TILs are a crucial component of cell-mediated immunity. The significance of CD8+ TILs has not been reported respectively in organ- and non-organ-confined urothelial carcinoma (UC) of the bladder. We explored the prognostic value of CD8+ TILs in the two groups. The presence of CD8+ TILs was assessed by immunohistochemical staining of whole tissue sections from 75 organ and 51 non-organ-confined disease patients with long-term follow-up, and its correlation with clinicopathological features and overall survival (OS) was determined. The CD8+ TIL immunohistochemical staining score was 0 (<1%), 1 (≥1%), 2 (≥5%), or 3 (≥10%) based on the percentage of positively stained cells out of total cells. A patient was considered CD8 negative if the score was 0. There were no associations between CD8+ TILs and age, sex, nuclear grade, and adjuvant or neoadjuvant chemotherapy in organ- and non-organ-confined disease. The presence of CD8+ TILs was seen more frequently in pTa-1 than pT2 stage (p = 0.033) in organ-confined disease. No associations between CD8+ TILs and pT stage, pN stage were found in non-organ-confined disease. CD8+ TILs were associated with better OS (log-rank test, P = 0.036) in non-organ-confined disease, but with poorer OS (log-rank test, P = 0.040) in organ-confined disease by the Kaplan–Meier method. In multivariate analysis, CD8+ TILs were an independent favorable prognostic factor in non-organ-confined disease, but were an independent unfavorable prognostic factor in organ-confined disease. These results suggest that CD8+ TILs have clinically significant anti-tumor activity in non-organ-confined disease, but may have pro-tumor activity in organ-confined disease. Therefore, we should be cautious if CD8+ TILs are aimed to be exploited in the treatment of bladder cancer.
Zhang et al. (2017) Tumor-infiltrating CD8+ lymphocytes predict different clinical outcomes in organ- and non-organ-confined urothelial carcinoma of the bladder following radical cystectomy. PeerJ 5:e3921 https://doi.org/10.7717/peerj.3921
A retrospective study: does upper airway morphology differ between non-positional and positional obstructive sleep apnea?
Objective The objective of this study was to explore the differences in upper airway morphology between positional (POSA) and non-positional (NPOSA) obstructive sleep apnea. Methods This retrospective study enrolled 75 patients (45 NPOSA and 30 POSA) who underwent polysomnography (PSG) and computed tomography (CT). The differences in, and relationships of, the PSG values and CT data between POSA and NPOSA were analyzed. Results Significant (p < 0.05) differences between the two groups were found in the apnea/hypopnea index (AHI), lateral-AHI (L-AHI), soft palate length (SPL), cross-sectional palatopharyngeal area, and the coronal diameter (CD) of the palatopharyngeal area at the narrowest part of the glossopharynx, which were all higher in POSA, except for SPL, AHI, and L-AHI. L-AHI was correlated with the cross-sectional area (r = − 0.306, p = 0.008) and CD (r = − 0.398, p < 0.001) of the palatopharyngeal area, the cross-sectional area (r = − 0.241, p = 0.038) and CD (r = − 0.297, p = 0.010) of the narrowest level of the glossopharynx, the CD of the glossopharynx (r = 0.284, p = 0.013), body mass index (BMI, r = 0.273, p = 0.018), SPL (r = 0.284, p = 0.014), and vallecula-tip of tongue (r = 0.250, p = 0.030). The SPL and CD at the narrowest part of the glossopharynx were included in the simplified screening model. Conclusions In NPOSA, the CD of the upper airway was smaller, and the soft palate was longer, than in POSA. These differences may play significant roles in explaining the main differences between NPOSA and POSA.
Jiao et al. (2017) A retrospective study: does upper airway morphology differ between non-positional and positional obstructive sleep apnea?. PeerJ 5:e3918 https://doi.org/10.7717/peerj.3918
Neurodynamic mobilization and foam rolling improved delayed-onset muscle soreness in a healthy adult population: a randomized controlled clinical trial
Objectives Compare the immediate effects of a Neurodynamic Mobilization (NM) treatment or foam roller (FR) treatment after DOMS. Design Double blind randomised clinical trial. Setting The participants performed 100 drop jumps (5 sets of 20 repetitions, separated by 2 min rests) from a 0.5-m high box in a University biomechanics laboratory to induce muscle soreness. The participants were randomly assigned in a counter-balanced fashion to either a FR or NM treatment group. Participants Thirty-two healthy subjects (21 males and 11 females, mean age 22.6 ± 2.2 years) were randomly assigned into the NM group (n = 16) or the FR group (n = 16). Main Outcome Measures The numeric pain rating scale (NPRS; 0–10), isometric leg strength with dynamometry, surface electromyography at maximum voluntary isometric contraction (MVIC) and muscle peak activation (MPA) upon landing after a test jump were measured at baseline, 48 h after baseline before treatment, and immediately after treatment. Results Both groups showed significant reduction in NPRS scores after treatment (NM: 59%, p < .01; FR: 45%, p < .01), but no difference was found between them (p > .05). The percentage change improvement in the MVIC for the rectus femoris was the only significant difference between the groups (p < 0.05) at post-treatment. After treatment, only the FR group had a statistically significant improvement (p < 0.01) in strength compared to pre-treatment. Conclusion Our results illustrate that both treatments are effective in reducing pain perception after DOMS whereas only FR application showed differences for the MVIC in the rectus femoris and strength.
Romero-Moraleda et al. (2017) Neurodynamic mobilization and foam rolling improved delayed-onset muscle soreness in a healthy adult population: a randomized controlled clinical trial. PeerJ 5:e3908 https://doi.org/10.7717/peerj.3908
Multigene phylogeny of the scyphozoan jellyfish family Pelagiidae reveals that the common U.S. Atlantic sea nettle comprises two distinct species (Chrysaora quinquecirrha and C. chesapeakei)
Background Species of the scyphozoan family Pelagiidae (e.g., Pelagia noctiluca, Chrysaora quinquecirrha) are well-known for impacting fisheries, aquaculture, and tourism, especially for the painful sting they can inflict on swimmers. However, historical taxonomic uncertainty at the genus (e.g., new genus Mawia) and species levels hinders progress in studying their biology and evolutionary adaptations that make them nuisance species, as well as ability to understand and/or mitigate their ecological and economic impacts. Methods We collected nuclear (28S rDNA) and mitochondrial (cytochrome c oxidase I and 16S rDNA) sequence data from individuals of all four pelagiid genera, including 11 of 13 currently recognized species of Chrysaora. To examine species boundaries in the U.S. Atlantic sea nettle Chrysaora quinquecirrha, specimens were included from its entire range along the U.S. Atlantic and Gulf of Mexico coasts, with representatives also examined morphologically (macromorphology and cnidome). Results Phylogenetic analyses show that the genus Chrysaora is paraphyletic with respect to other pelagiid genera. In combined analyses, Mawia, sampled from the coast of Senegal, is most closely related to Sanderia malayensis, and Pelagia forms a close relationship to a clade of Pacific Chrysaora species (Chrysaora achlyos, Chrysaora colorata, Chrysaora fuscescens, and Chrysaora melanaster). Chrysaora quinquecirrha is polyphyletic, with one clade from the U.S. coastal Atlantic and another in U.S. Atlantic estuaries and Gulf of Mexico. These genetic differences are reflected in morphology, e.g., tentacle and lappet number, oral arm length, and nematocyst dimensions. Caribbean sea nettles (Jamaica and Panama) are genetically similar to the U.S. Atlantic estuaries and Gulf of Mexico clade of Chrysaora quinquecirrha. Discussion Our phylogenetic hypothesis for Pelagiidae contradicts current generic definitions, revealing major disagreements between DNA-based and morphology-based phylogenies. A paraphyletic Chrysaora raises systematic questions at the genus level for Pelagiidae; accepting the validity of the recently erected genus Mawia, as well as past genera, will require the creation of additional pelagiid genera. Historical review of the species-delineating genetic and morphological differences indicates that Chrysaora quinquecirrha Desor 1848 applies to the U.S. Coastal Atlantic Chrysaora species (U.S. Atlantic sea nettle), while the name C. chesapeakei Papenfuss 1936 applies to the U.S. Atlantic estuarine and Gulf of Mexico Chrysaora species (Atlantic bay nettle). We provide a detailed redescription, with designation of a neotype for Chrysaora chesapeakei, and clarify the description of Chrysaora quinquecirrha. Since Caribbean Chrysaora are genetically similar to Chrysaora chesapeakei, we provisionally term them Chrysaora c.f. chesapeakei. The presence of Mawia benovici off the coast of Western Africa provides a potential source region for jellyfish introduced into the Adriatic Sea in 2013.
Bayha et al. (2017) Multigene phylogeny of the scyphozoan jellyfish family Pelagiidae reveals that the common U.S. Atlantic sea nettle comprises two distinct species (Chrysaora quinquecirrha and C. chesapeakei). PeerJ 5:e3863 https://doi.org/10.7717/peerj.3863