Automatic detection of the parasite Trypanosoma cruzi in blood smears using a machine learning approach applied to mobile phone images
Chagas disease is a life-threatening illness caused by the parasite Trypanosoma cruzi. The diagnosis of the acute form of the disease is performed by trained microscopists who detect parasites in blood smear samples. Since this method requires a dedicated high-resolution camera system attached to the microscope, the diagnostic method is more expensive and often prohibitive for low-income settings. Here, we present a machine learning approach based on a random forest (RF) algorithm for the detection and counting of T. cruzi trypomastigotes in mobile phone images. We analyzed micrographs of blood smear samples that were acquired using a mobile device camera capable of capturing images in a resolution of 12 megapixels. We extracted a set of features that describe morphometric parameters (geometry and curvature), as well as color, and texture measurements of 1,314 parasites. The features were divided into train and test sets (4:1) and classified using the RF algorithm. The values of precision, sensitivity, and area under the receiver operating characteristic (ROC) curve of the proposed method were 87.6%, 90.5%, and 0.942, respectively. Automating image analysis acquired with a mobile device is a viable alternative for reducing costs and gaining efficiency in the use of the optical microscope.
Morais et al. (2022) Automatic detection of the parasite Trypanosoma cruzi in blood smears using a machine learning approach applied to mobile phone images. PeerJ 10:e13470 https://doi.org/10.7717/peerj.13470
Proteomic analysis of differential anther development from sterile/fertile lines in Capsicum annuum L.
Background Pepper (Capsicum annuum L.) is a major cash crop throughout the world. Male sterility is an important characteristic in crop species that leads to a failure to produce functional pollen, and it has crucial roles in agricultural breeding and the utilization of heterosis. Objectives In this study, we identified many crucial factors and important components in metabolic pathways in anther and pollen development, and elucidated the molecular mechanism related to pollen abortion in pepper. Methods Pepper pollen was observed at different stages to detect the characteristics associated with male sterility and fertility. The phytohormone and oxidoreductase activities were detected in spectrophotometric and redox reaction assays, respectively. Proteins were extracted from male sterile and fertile pepper lines, and identified by TMT/iTRAQ (tandem mass tags/isobaric tags for relative and absolute quantitation) and LC-MS/MS (liquid chromatograph-mass spectrometer) analysis. Differentially abundant proteins (DAPs) were analyzed based on Gene Ontology annotations and the Kyoto Encyclopedia of Genes and Genomes database according to |fold change)| > 1.3 and P value < 0.05. DAPs were quantified in the meiosis, tetrad, and binucleate stages by parallel reaction monitoring (PRM). Results In this study, we screened and identified one male sterile pepper line with abnormal cytological characteristics in terms of pollen development. The peroxidase and catalase enzyme activities were significantly reduced and increased, respectively, in the male sterile line compared with the male fertile line. Phytohormone analysis demonstrated that the gibberellin, jasmonic acid, and auxin contents changed by different extents in the male sterile pepper line. Proteome analysis screened 1,645 DAPs in six clusters, which were mainly associated with the chloroplast and cytoplasm based on their similar expression levels. According to proteome analysis, 45 DAPs were quantitatively identified in the meiosis, tetrad, and binucleate stages by PRM, which were related to monoterpenoid biosynthesis, and starch and sucrose metabolism pathways. Conclusions We screened 1,645 DAPs by proteomic analysis and 45 DAPs were related to anther and pollen development in a male sterile pepper line. In addition, the activities of peroxidase and catalase as well as the abundances of phytohormones such as gibberellin, jasmonic acid, and auxin were related to male sterility. The results obtained in this study provide insights into the molecular mechanism responsible for male sterility and fertility in pepper.
Pei et al. (2022) Proteomic analysis of differential anther development from sterile/fertile lines in Capsicum annuum L.. PeerJ 10:e13168 https://doi.org/10.7717/peerj.13168
Ability of functional performance assessments to discriminate athletes with and without chronic ankle instability : a case-control study
Background The decline in motor function associated with chronic ankle instability (CAI) can be assessed using Functional Performance tests. Ankle muscular strength, endurance and range of motion (ROM) has been assessed in previous studies but functional activities such as sprinting and change of direction are less well studied in athletes with CAI. Hence the aim of this study was to determine how sprint, change of direction, ankle isometric strength, endurance and ROM measures may be associated with discriminate athletes with and without CAI. Methods One hundred and six participants (CAI: n = 53 or no CAI: n = 53) provided informed consent to participate in this study. Participants performed three functional performance tests, (30-m sprint test, Modified Illinois change of direction test (MICODT)) and change of direction test. Range of motion for dorsiflexion was measured using weight bearing lunge test and inversion, eversion and plantarflexion using Saunders® digital inclinometer. Strength was assessed using Baseline® hand-held dynamometer for plantarflexors, dorsiflexors, invertors and evertors. Muscular endurance was assessed by single heel raise test and Modified single heel raise test. Between-group comparisons utilised Student’s t-test and Mann-Whitney U-tests, with a number of unique variable and multivariable binomial logistic regression performed to determine which performance measures may discriminate participants with CAI. Results The CAI participants performed significantly worse in the three functional performance tests as well as multiple measures of ankle ROM, isometric strength and muscular endurance (p < 0.008). While several measures of ROM (plantarflexion and dorsiflexion), strength (inversion and eversion) and both muscular endurance tests were significantly associated with CAI in the univariable analysis, the strongest association was the functional performance tests, especially MICDOT time (odds ratio (95% CI): 0.06 [0.02–0.17], sensitivity 94.3%, specificity 88.7%). Multivariable regression analyses indicated that performance across the functional performance tests were more strongly associated with CAI than any ankle ROM, muscular strength or endurance test. Further, the inclusion of the best ankle range of motion, strength or muscular endurance tests did not significantly improve upon the association of the MICDOT with CAI. Conclusions Chronic ankle instability in athletic populations appears to be highly associated with declines in functional performance and to a somewhat lesser extent, ankle range of motion, strength and muscle endurance measures. This may suggest that optimal rehabilitation for athletes with CAI may require a greater focus on improving sprinting speed and change of direction ability in the mid to latter stages of rehabilitation, with regular assessments of these functional performance tests necessary to guide the progression and overload of this training.
S. Jamsandekar et al. (2022) Ability of functional performance assessments to discriminate athletes with and without chronic ankle instability : a case-control study. PeerJ 10:e13390 https://doi.org/10.7717/peerj.13390
Challenges of sperm cryopreservation in transferring heat adaptation of corals across ocean basins
Reef-building corals live very close to their upper thermal limits and their persistence is imperiled by a rapidly warming climate. Human interventions may be used to increase the thermal limits of sensitive corals by cross-breeding with heat-adapted populations. However, the scope of breeding interventions is constrained by regional variation in the annual reproductive cycle of corals. Here we use cryopreservation technology to overcome this barrier and cross-breed conspecific coral populations across ocean basins for the first time. During regional spawning events, sperm samples were cryopreserved from populations of the widespread Indo-Pacific coral, Platygyra daedalea, from the southern Persian Gulf (maximum daily sea surface temperature of 36 °C), the Oman Sea (33 °C), and the central Great Barrier Reef (30 °C). These sperm samples were thawed during a later spawning event to test their ability to fertilize freshly spawned eggs of P. daedalea colonies from the central Great Barrier Reef. Average fertilization success for the Persian Gulf (9%) and Oman Sea (6%) sperm were 1.4–2.5 times lower than those for the native cryopreserved sperm from Great Barrier Reef (13–15%), potentially due to lower sperm quality of the Middle Eastern sperm and/or reproductive incompatibility between these distant populations. Overall, fertilization success with cryopreserved sperm was low compared with fresh sperm (>80%), likely due to the low motility of thawed sperm (≤5%, reduced from 50% to >90% in fresh sperm). To evaluate whether cross-bred offspring had enhanced thermal tolerance, the survival of larvae sired by Persian Gulf cryopreserved sperm, Great Barrier Reef cryopreserved sperm, and Great Barrier Reef fresh sperm was monitored for six days at ambient (27 °C) and elevated (33 °C) temperature. Against expectations of thermal tolerance enhancement, survival of larvae sired by Persian Gulf cryopreserved sperm was 2.6 times lower than larvae sired by Great Barrier Reef fresh sperm at 33 °C (27% versus 71%), but did not differ at 27 °C (77% versus 84%). This lack of enhanced thermal tolerance was unlikely due to outbreeding depression as survival was equally poor in larvae sired by Great Barrier Reef cryopreserved sperm. Rather, follow-up tests showed that cryoprotectant exposure during fertilization (0.1% DMSO) has a negative effect on the survival of P. daedalea larvae which is exacerbated at elevated temperature. Collectively, our findings highlight challenges of breeding corals for enhanced thermal tolerance using cryopreserved sperm, which may be overcome by methodological advances in the collection and preservation of high-quality motile sperm and minimizing the exposure time of eggs to cryoprotectants.
Howells et al. (2022) Challenges of sperm cryopreservation in transferring heat adaptation of corals across ocean basins. PeerJ 10:e13395 https://doi.org/10.7717/peerj.13395
Influence of DNA extraction kits on freshwater fungal DNA metabarcoding
Background Environmental DNA (eDNA) metabarcoding is a common technique for efficient biodiversity monitoring, especially of microbes. Recently, the usefulness of aquatic eDNA in monitoring the diversity of both terrestrial and aquatic fungi has been suggested. In eDNA studies, different experimental factors, such as DNA extraction kits or methods, can affect the subsequent analyses and the results of DNA metabarcoding. However, few methodological studies have been carried out on eDNA of fungi, and little is known about how experimental procedures can affect the results of biodiversity analysis. In this study, we focused on the effect of DNA extraction method on fungal DNA metabarcoding using freshwater samples obtained from rivers and lakes. Methods DNA was extracted from freshwater samples using the DNeasy PowerSoil kit, which is mainly used to extractmicrobial DNA from soil, and the DNeasy Blood & Tissue kit, which is commonly used for eDNA studies on animals. We then compared PCR inhibition and fungal DNA metabarcoding results; i.e., operational taxonomic unit (OTU) number and composition of the extracted samples. Results No PCR inhibition was detected in any of the samples, and no significant differences in the number of OTUs and OTU compositions were detected between the samples processed using different kits. These results indicate that both DNA extraction kits may provide similar diversity results for the river and lake samples evaluated in this study. Therefore, it may be possible to evaluate the diversity of fungi using a unified experimental method, even with samples obtained for diversity studies on other taxa such as those of animals.
Matsuoka et al. (2022) Influence of DNA extraction kits on freshwater fungal DNA metabarcoding. PeerJ 10:e13477 https://doi.org/10.7717/peerj.13477
Sexual dimorphism in shell size of the land snail Leptopoma perlucidum (Caenogastropoda: Cyclophoridae)
Sexual dimorphism in the shell size and shape of land snails has been less explored compared to that of other marine and freshwater snail taxa. This study examined the differences in shell size and shape across both sexes of Leptopoma perlucidum land snails. We collected 84 land snails of both sexes from two isolated populations on two islands off Borneo. A total of five shell size variables were measured: (1) shell height, (2) shell width, (3) shell spire height, (4) aperture height, and (5) aperture width. We performed frequentist and Bayesian t-tests to determine if there was a significant difference between the two sexes of L. perlucidum on each of the five shell measurements. Additionally, the shell shape was quantified based on nine landmark points using the geometric morphometric approach. We used generalised Procrustes and principal component analyses to test the effects of sex and location on shell shape. The results showed that female shells were larger than male shells across all five measurements (all with p-values < 0.05), but particularly in regards to shell height and shell width. Future taxonomic studies looking to resolve the Leptopoma species’ status should consider the variability of shell size caused by sexual dimorphism.
Phung et al. (2022) Sexual dimorphism in shell size of the land snail Leptopoma perlucidum (Caenogastropoda: Cyclophoridae). PeerJ 10:e13501 https://doi.org/10.7717/peerj.13501
Anticancer mechanism of 7-α-hydroxyfrullanolide on microtubules and computational prediction of its target binding in triple-negative breast cancer cells
Background Triple-negative breast cancer (TNBC) responds poorly to the available drugs; thus, the mortality rate associated with TNBC remains high. 7-α-Hydroxyfrullanolide (7HF) possesses anticancer properties and arrests cells in the G2/M-phase via modulation of several proteins involved in the G2/M-phase transition, as well as the mitotic checkpoint in MDA-MB-468 (TNBC) cells. Microtubules (MTs) dynamically regulate cell division in the G2/M phase and are related to cancer cell stress response. However, antimitotic drug cytotoxicity to multiple cancer resistance developed in response to drugs are obstacles faced to date. Here, the activity and mechanism via which 7HF controls MTs dynamics was investigated in MDA-MB-468 cells. Methods 7HF uptake by MDA-MB-468 cells was assessed using spectrophotometry. The drug-like properties of 7HF were predicted using the Swiss-absorption, distribution, metabolism, and excretion (ADME) webtool. Then, the effect of 7HF treatment (6, 12, and 24 µM) on the dynamic arrangement of MTs was assessed for 1, 12, and 24 h using indirect immunofluorescence. Polymerization of α- and β-tubulin was assessed using different 7HF concentrations in a cell-free system for 1 h. Cell proliferation assay with bromodeoxyuridine plus propidium iodide staining and flow cytometry was performed at different 7HF concentrations and time points. The mechanism of action was assessed by detecting the expression of proteins, including Bub3, cyclin B1, p-Cdk1 (Tyr15), Rb, p-Rb (Ser780), Chk1, p-Chk1 (Ser345), Chk2, p-Chk2 (Ser516), and p-H2AX (Ser139), using western blotting. Molecular docking was used to predict the molecular interactions between 7HF and tubulins in MTs. Results We observed that 7HF was able to enter the MDA-MB-468 cells. The ADME webtool analysis predicted that it possesses the high passive permeation and gastrointestinal absorption properties of drugs. Various concentrations of 7HF disrupted the dynamic arrangement of spindle MTs by causing radial spindle array shrinkage and expansion of fibrous spindle density and radial array lengths in a time-dependent manner. 7HF reduced polymerization of α-, β-tubulin in dose-dependent manner. 7HF also triggered DNA damage response by inducing G2/M and G1 phase arrests in a concentration and time-dependent manner, which occurred due to the upregulation of Bub3, Chk1, p-Chk1 (Ser345), p-Cdk1 (Tyr15), and cyclin B1. According to molecular docking analysis, 7HF preferred to bind to β-tubulin over α-tubulin. The lactone, ketone, and hydroxyl groups of 7HF supported the 7HF-tubulin interactions. Hydrogen bonding with a hydrocarbon ring and salt bridge attractive forces were responsible for the binding versatility of 7HF. Conclusions This is the first study to investigate the molecular mechanism, MTs interacting sites, and the internalization and drug-like properties of 7HF in TNBC cells. The findings will be useful for developing 7HF-based treatment for patients with TNBC.
Chimplee et al. (2022) Anticancer mechanism of 7-α-hydroxyfrullanolide on microtubules and computational prediction of its target binding in triple-negative breast cancer cells. PeerJ 10:e13508 https://doi.org/10.7717/peerj.13508
Predicting habitat suitability and range shifts under projected climate change for two octocorals in the north-east Atlantic
Species distribution models have become a valuable tool to predict the distribution of species across geographic space and time. In this study, maximum entropy models were constructed for two temperate shallow-water octocoral species, the pink sea fan (Eunicella verrucosa) and dead man’s fingers (Alcyonium digitatum), to investigate and compare habitat suitability. The study area covered the north-east Atlantic from the Bay of Biscay to the British Isles and southern Norway; this area includes both the northern range of E. verrucosa and the middle-northern range of A. digitatum. The optimal models for each species showed that, overall, slope, temperature at the seafloor and wave orbital velocity were important predictors of distribution in both species. Predictions of habitat suitability showed areas of present-day (1951–2000) suitable habitat where colonies have not yet been observed, particularly for E. verrucosa, where areas beyond its known northern range limit were identified. Moreover, analysis with future layers (2081–2100) of temperature and oxygen concentration predicted a sizable increase in habitat suitability for E. verrucosa beyond these current range limits under the Representative Concentration Pathway 8.5 scenario. This suggests that projected climate change may induce a potential range expansion northward for E. verrucosa, although successful colonisation would also be conditional on other factors such as dispersal and interspecific competition. For A. digitatum, this scenario of projected climate change may result in more suitable habitat in higher latitudes, but, as with E. verrucosa, there is a degree of uncertainty in the model predictions. Importantly, the results from this study highlight present-day areas of high habitat suitability which, if combined with knowledge on population density, could be used to identify priority areas to enhance protection and ensure the long-term survival of these octocoral species in the region.
Jenkins et al. (2022) Predicting habitat suitability and range shifts under projected climate change for two octocorals in the north-east Atlantic. PeerJ 10:e13509 https://doi.org/10.7717/peerj.13509
Animal reactivity to camera traps and its effects on abundance estimate using distance sampling in the Taï National Park, Côte d’Ivoire
The use of camera traps (CTs) has become an increasingly popular method of studying wildlife, as CTs are able to detect rare, nocturnal, and elusive species in remote and difficult-to-access areas. It thus makes them suited to estimate animal density and abundance, identify activity patterns and new behaviours of animals. However, animals can react when they see the CTs and this can lead to bias in the animal population estimates. While CTs may provide many advantages, an improved understanding of their impacts on individual’s behaviour is necessary to avoid erroneous density estimates. Yet, the impact of CTs on detected individuals, such as human odour near the device and the environment, or the infrared illumination, has received relatively little attention. To date, there is no clear procedure to remove this potential bias. Here, we use camera trap distance sampling (CTDS) to (1) quantify the bias resulting from the different animal responses to the CTs when determining animal density and abundance, and (2) test if olfactory, visual and auditory signals have an influence on the animals’ reaction to CTs. Between March 2019 and March 2020, we deployed CTs at 267 locations distributed systematically over the entire Taï National Park. We obtained 58,947 videos from which we analysed four medium- to-large-bodied species (Maxwell’s duiker (Philantomba maxwellii), Jentink’s duiker (Cephalophus jentinki), pygmy hippopotamus (Choeropsis liberiensis) and Western chimpanzee (Pan troglodytes verus)) displaying different behaviours towards the CTs. We then established species-specific ethograms describing the behavioural responses to the CTs. Using these species-specific responses, we observed that the Maxwell’s duiker reacted weakly to CTs (about 0.11% of the distance data), contrary to Jentink’s duiker, pygmy hippopotamus and Western chimpanzee which reacted with relatively high frequencies, representing 32.82%, 52.96% and 16.14% of the distance data, respectively. Not taking into account the species-specific responses to the CTs can lead to an artificial doubling or tripling of the populations’ sizes. All species reacted more to the CTs at close distances. Besides, the Jentink’s duiker and the pygmy hippopotamus reacted significantly more to the CTs at night than during the day. Finally, as for olfactory signals, the probability of reaction to the CTs during the first days after CTs installation was weak in Maxwell’s duiker, but concerned 18% of the video captures in Western chimpanzees which decreasing with time, but they remained high in pygmy hippopotamus and Jentink’s duiker (65% and 70% of the video captures respectively). Careful consideration should be given to animal’s response to CTs during the analysis and in the field, by reducing human’s impact around the CTs installation.
Houa et al. (2022) Animal reactivity to camera traps and its effects on abundance estimate using distance sampling in the Taï National Park, Côte d’Ivoire. PeerJ 10:e13510 https://doi.org/10.7717/peerj.13510
Trajectory patterns and factors influencing perinatal fatigue among Chinese women from late pregnancy to 6 months after delivery
Background Perinatal fatigue among women is related to the clinical outcomes of mothers and infants. Perinatal fatigue changes over time, and the trajectory varies according to the predictors and outcomes of the mothers. This, however, has not been documented in any study. Objectives This study aimed to identify and characterize the trajectory patterns of perinatal fatigue among women from late pregnancy to 6 months after delivery. Methods We used growth mixture modeling to estimate the trajectory of perinatal fatigue at 28 gestational weeks (T0), 37 gestational weeks (T1), 3 days (T2), 1 week (T3), 6 weeks (T4), and 6 months (T5) after delivery with (n = 1,030). The Mann-Whitney U test and binary logistic regression were used to tie the selected trajectory classes to predictors and outcomes. Results There were two distinct patterns of perinatal fatigue in women: “persistently high” (11.1%, n = 114) and “persistently low” (88.9%, n = 916). Levels of perinatal fatigue among women in the “persistently high” group were higher than those in the “persistently low” group across the six measurements. Complications, fatigue at T0, and employment status in late pregnancy were all significant predictors of trajectories. Additionally, the “persistently high” group had a greater prevalence of difficult baby care and weight retention and a lower prevalence of exclusive breastfeeding. Conclusions Our study proved the heterogeneity and characteristics of perinatal fatigue among women. Future research should concentrate on developing intervention packages targeted at specific individuals in order to alleviate perinatal fatigue in women.
Zhu et al. (2022) Trajectory patterns and factors influencing perinatal fatigue among Chinese women from late pregnancy to 6 months after delivery. PeerJ 10:e13387 https://doi.org/10.7717/peerj.13387