Structural modeling and functional analysis of the essential ribosomal processing protease Prp from Staphylococcus aureus

In Firmicutes and related bacteria, ribosomal large subunit protein L27 is encoded with a conserved N-terminal extension that is removed to expose residues critical for ribosome function. Bacteria encoding L27 with this N-terminal extension also encode a sequence-specific cysteine protease, Prp, which carries out this cleavage. In this work, we demonstrate that L27 variants with an un-cleavable N-terminal extension, or lacking the extension (pre-cleaved), are unable to complement an L27 deletion in Staphylococcus aureus. This indicates that N-terminal processing of L27 is not only essential but possibly has a regulatory role. Prp represents a new clade of previously uncharacterized cysteine proteases, and the dependence of S. aureus on L27 cleavage by Prp validates the enzyme as a target for potential antibiotic development. To better understand the mechanism of Prp activity, we analyzed Prp enzyme kinetics and substrate preference using a fluorogenic peptide cleavage assay. Molecular modeling and site-directed mutagenesis implicate several residues around the active site in catalysis and substrate binding, and support a structural model in which rearrangement of a flexible loop upon binding of the correct peptide substrate is required for the active site to assume the proper conformation. These findings lay the foundation for the development of antimicrobials that target this novel, essential pathway

A mixed-model moving-average approach to geostatistical modeling in stream networks

Spatial autocorrelation is an intrinsic characteristic in freshwater stream environments where nested watersheds and flow connectivity may produce patterns that are not captured by Euclidean distance. Yet, many common autocovariance functions used in geostatistical models are statistically invalid when Euclidean distance is replaced with hydrologic distance. We use simple worked examples to illustrate a recently developed moving-average approach used to construct two types of valid autocovariance models that are based on hydrologic distances. These models were designed to represent the spatial configuration, longitudinal connectivity, discharge, and flow direction in a stream network. They also exhibit a different covariance structure than Euclidean models and represent a true difference in the way that spatial relationships are represented. Nevertheless, the multi-scale complexities of stream environments may not be fully captured using a model based on one covariance structure. We advocate using a variance component approach, which allows a mixture of autocovariance models (Euclidean and stream models) to be incorporated into a single geostatistical model. As an example, we fit and compare ‘‘mixed models,’’ based on multiple covariance structures, for a biological indicator. The mixed model proves to be a flexible approach because many sources of information can be incorporated into a single model

Correcting misclassification errors in crowdsourced ecological data: A Bayesian perspective

Many research domains use data elicited from "citizen scientists" when a direct measure of a process is expensive or infeasible. However, participants may report incorrect estimates or classifications due to their lack of skill. We demonstrate how Bayesian hierarchical models can be used to learn about latent variables of interest, while accounting for the participants' abilities. The model is described in the context of an ecological application that involves crowdsourced classifications of georeferenced coral-reef images from the Great Barrier Reef, Australia. The latent variable of interest is the proportion of coral cover, which is a common indicator of coral reef health. The participants' abilities are expressed in terms of sensitivity and specificity of a correctly classified set of points on the images. The model also incorporates a spatial component, which allows prediction of the latent variable in locations that have not been surveyed. We show that the model outperforms traditional weighted-regression approaches used to account for uncertainty in citizen science data. Our approach produces more accurate regression coefficients and provides a better characterization of the latent process of interest. This new method is implemented in the probabilistic programming language Stan and can be applied to a wide number of problems that rely on uncertain citizen science data.Comment: 18 figures, 5 table

Socio-Cultural, Organizational, and Community Level Influences on Physical Activity Levels of Latino Preschool-Age Children: A Qualitative Study

Objectives: As more children grow up in families with immigrant parents of Latino origin, there is a need to understand key influences on physical activity behaviors of young Latino children to prevent obesity in this high-risk group.Design: We conducted six focus groups with low-income Latina mothers (N = 33) whose preschool-aged children (2-5 years) were enrolled in the Supplemental Nutrition Program for Women, Infants and Children (WIC) program in Rhode Island. Data was analyzed using content analysis to identify recurrent themes.Results: Despite understanding the importance of physical activity for overall health, physical activity was not a top priority for the Latino mothers participating in the focus groups. Mothers reported facing numerous barriers to establishing and maintaining healthful physical activity habits for their preschool-aged children and themselves, particularly financial and socio-cultural barriers. Analyses revealed that Latina mothers perceive the WIC as a program focused on the development and maintenance of healthy eating habits and nutritional status of children and not physical activity.Conclusions: Recognizing the importance of socioeconomic position and the influence of cultural factors on physical activity is essential if effective prevention and intervention programs for Latino families and their children are to be designed. Study findings emphasize the importance of the family as a central unit of change and suggest that successful interventions to promote physical activity of low-income Latino preschool children must take into account the needs and constraints of the family unit as a whole. The WIC program has the potential to be a venue for promoting awareness of and educating low-income Latino parents about the importance of helping their children develop and maintain early healthful physical activity habits. The WIC program can also play an important role in facilitating access and creating programs and services that provide increased opportunities for physical activity of young children and their families

Bayesian spatio-temporal models for stream networks

Spatio-temporal models are widely used in many research areas including ecology. The recent proliferation of the use of in-situ sensors in streams and rivers supports space-time water quality modelling and monitoring in near real-time. In this paper, we introduce a new family of dynamic spatio-temporal models, in which spatial dependence is established based on stream distance and temporal autocorrelation is incorporated using vector autoregression approaches. We propose several variations of these novel models using a Bayesian framework. Our results show that our proposed models perform well using spatio-temporal data collected from real stream networks, particularly in terms of out-of-sample RMSPE. This is illustrated considering a case study of water temperature data in the northwestern United States.Comment: 26 pages, 10 fig

Increasing trust in new data sources: crowdsourcing image classification for ecology

Crowdsourcing methods facilitate the production of scientific information by non-experts. This form of citizen science (CS) is becoming a key source of complementary data in many fields to inform data-driven decisions and study challenging problems. However, concerns about the validity of these data often constrain their utility. In this paper, we focus on the use of citizen science data in addressing complex challenges in environmental conservation. We consider this issue from three perspectives. First, we present a literature scan of papers that have employed Bayesian models with citizen science in ecology. Second, we compare several popular majority vote algorithms and introduce a Bayesian item response model that estimates and accounts for participants' abilities after adjusting for the difficulty of the images they have classified. The model also enables participants to be clustered into groups based on ability. Third, we apply the model in a case study involving the classification of corals from underwater images from the Great Barrier Reef, Australia. We show that the model achieved superior results in general and, for difficult tasks, a weighted consensus method that uses only groups of experts and experienced participants produced better performance measures. Moreover, we found that participants learn as they have more classification opportunities, which substantially increases their abilities over time. Overall, the paper demonstrates the feasibility of CS for answering complex and challenging ecological questions when these data are appropriately analysed. This serves as motivation for future work to increase the efficacy and trustworthiness of this emerging source of data.Comment: 25 pages, 10 figure

Identification of a transporter complex responsible for the cytosolic entry of nitrogen-containing bisphosphonates

Nitrogen-containing-bisphosphonates (N-BPs) are widely prescribed to treat osteoporosis and other bone-related diseases. Although previous studies established that N-BPs function by inhibiting the mevalonate pathway in osteoclasts, the mechanism by which N-BPs enter the cytosol from the extracellular space to reach their molecular target is not understood. Here we implemented a CRISPRi-mediated genome-wide screen and identified SLC37A3 (solute carrier family 37 member A3) as a gene required for the action of N-BPs in mammalian cells. We observed that SLC37A3 forms a complex with ATRAID (all-trans retinoic acid-induced differentiation factor), a previously identified genetic target of N-BPs. SLC37A3 and ATRAID localize to lysosomes and are required for releasing N-BP molecules that have trafficked to lysosomes through fluid-phase endocytosis into the cytosol. Our results elucidate the route by which N-BPs are delivered to their molecular target, addressing a key aspect of the mechanism of action of N-BPs that may have significant clinical relevance

A framework for automated anomaly detection in high frequency water-quality data from in situ sensors

River water-quality monitoring is increasingly conducted using automated in situ sensors, enabling timelier identification of unexpected values. However, anomalies caused by technical issues confound these data, while the volume and velocity of data prevent manual detection. We present a framework for automated anomaly detection in high-frequency water-quality data from in situ sensors, using turbidity, conductivity and river level data. After identifying end-user needs and defining anomalies, we ranked their importance and selected suitable detection methods. High priority anomalies included sudden isolated spikes and level shifts, most of which were classified correctly by regression-based methods such as autoregressive integrated moving average models. However, using other water-quality variables as covariates reduced performance due to complex relationships among variables. Classification of drift and periods of anomalously low or high variability improved when we applied replaced anomalous measurements with forecasts, but this inflated false positive rates. Feature-based methods also performed well on high priority anomalies, but were also less proficient at detecting lower priority anomalies, resulting in high false negative rates. Unlike regression-based methods, all feature-based methods produced low false positive rates, but did not and require training or optimization. Rule-based methods successfully detected impossible values and missing observations. Thus, we recommend using a combination of methods to improve anomaly detection performance, whilst minimizing false detection rates. Furthermore, our framework emphasizes the importance of communication between end-users and analysts for optimal outcomes with respect to both detection performance and end-user needs. Our framework is applicable to other types of high frequency time-series data and anomaly detection applications

Using virtual reality and thermal imagery to improve statistical modelling of vulnerable and protected species.

Biodiversity loss and sparse observational data mean that critical conservation decisions may be based on little to no information. Emerging technologies, such as airborne thermal imaging and virtual reality, may facilitate species monitoring and improve predictions of species distribution. Here we combined these two technologies to predict the distribution of koalas, specialized arboreal foliovores facing population declines in many parts of eastern Australia. For a study area in southeast Australia, we complemented ground-survey records with presence and absence observations from thermal-imagery obtained using Remotely-Piloted Aircraft Systems. These field observations were further complemented with information elicited from koala experts, who were immersed in 360-degree images of the study area. The experts were asked to state the probability of habitat suitability and koala presence at the sites they viewed and to assign each probability a confidence rating. We fit logistic regression models to the ground survey data and the ground plus thermal-imagery survey data and a Beta regression model to the expert elicitation data. We then combined parameter estimates from the expert-elicitation model with those from each of the survey models to predict koala presence and absence in the study area. The model that combined the ground, thermal-imagery and expert-elicitation data substantially reduced the uncertainty around parameter estimates and increased the accuracy of classifications (koala presence vs absence), relative to the model based on ground-survey data alone. Our findings suggest that data elicited from experts using virtual reality technology can be combined with data from other emerging technologies, such as airborne thermal-imagery, using traditional statistical models, to increase the information available for species distribution modelling and the conservation of vulnerable and protected species