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The continental shelf of Brazil is home to a wide range of lobster species, with varying body size, color, habitat preference, and geographic and bathymetric distribution. Spiny lobsters (Panulirus) and slipper lobsters (Scyllaridae) are exploited for export and for the domestic market. Deep sea lobsters (Nephropidae and Polychelidae) have no commercial potential, and little is known about their biology. In this review, we identified 24 lobster species from benthic ecosystems off Brazil (Palinuridae 25%, Scyllaridae 29%, Nephropidae 25%, Polychelidae 17%, Enoplometopidae 4%). We designed a simplified theoretical scheme to understand the role of lobsters in the ecosystem, based on available evidence of distribution, biodiversity, life cycle, connectivity, and abundance. Finally, we propose a theoretical scheme of trophic top-down control, with interactions between a large decapod (spiny lobster), a demersal predator (red snapper), an apex predator (small tuna), benthic invertebrates and fishing exploitation.

Residency (R) and site fidelity (SF) are important parameters in population ecology, yet their quantification poses challenges in marine mammals. Based on a previous review, this study used simulated and empirical mark-resight data to assess the variations and performance of the most used R (n = 8) and SF (n = 11) indices in peer-reviewed literature under different scenarios. We applied the Jolly-Seber model to simulate thousands of bottlenose dolphin populations varying resighting (p) and survival (Phi) probabilities, and performed calibration, sensitivity, and validation analyses. Our results underscore the effects of p and Phi on individual categorization within the diverse simulated conditions, representing the often-overlooked heterogeneity in residency classification for Tursiops populations. All SF indices showed similar and consistent performance (>0.70 Gower’s distance) across the simulated scenarios, even when compared to field study data from wild dolphin populations (i.e., Savannah, USA, and Alvarado, Mexico); thus, SF should be a critical parameter for interstudy comparisons. Conversely, R indices were remarkably different based on their definitions and classification criteria. The different thresholds among definitions largely biased the proportion of residents and transient individuals (or occasional visitors) even leading to counterintuitive outcomes. This emphasizes the importance of considering trade-offs in R index selection aligned with project goals, specific sampling efforts, and population dynamics. For instance, the simplified binomial categorization of R defined by Conway (2017) (https://digitalcommons.coastal.edu/etd/10/) easier to interpret but R indices incorporating temporal components (e.g., monthly, seasonal, and annual) outperformed (>0.70 Gower’s distance) other R indices lacking such criteria. This allowed for a more detailed representation of the temporal structure of the population, and higher consistency and accuracy while classifying individuals. Also, although the residency categories proposed by Möller, Allen & Harcourt (2002) (DOI 10.1071/AM02011) did not perform as well, these seemed to fit better when dealing with data gaps across spatial and temporal scales. Our results contribute to the ongoing discussion on methodological implications for the interpretation of ecological patterns, facilitating a nuanced understanding of population dynamics, aiding scientists, and conservation agencies in making informed decisions for bottlenose dolphin populations worldwide.

Background The role of attentional focus is a well-explored topic in exercise sciences. Studies generally indicate that external focus (EF) enhances motor performance and learning compared with internal focus (IF). However, most studies only included one or two experimental days which limits participants’ exposure to the focus conditions. This raises the question of whether the superiority of EF varies over time. Methods Accordingly, in this pre-registered within-subject study, we examined the effects of focus instructions on golf-putting performance over four days, with 48–72 h between them. On each day, participants performed 15 putts under three instructional conditions: (1) EF, (2) IF, and (3) control, in a randomized and counterbalanced order. Results We observed trivial differences in performance between conditions but considerable improvements from day 1 to day 4. When using an exploratory analysis, we found that participants performed better under EF and control conditions compared with the IF condition on day 1, but not on subsequent days. Conclusions Since IF instructions are more commonly used in practice, we speculate that the two other focus conditions were experienced as more novel, potentially accounting for their superiority on Day 1. Nevertheless, our results question the significance of employing EF to enhance performance.

Background/Objectives: The incidence of acute kidney injury (AKI) has been steadily increasing. Despite its high prevalence, there is no pathogenetically rational therapy for AKI. This deficiency stems from the poor understanding of the pathogenesis of AKI. Renal ischemia/hypoxia is one of the leading causes of clinical AKI. This study investigates whether αMUPA mice, overexpressing the urokinase plasminogen activator (uPA) gene are protected against ischemic AKI, thus unraveling a potential renal damage treatment target. Methods: We utilized an in vivo model of I/R-induced AKI in αMUPA mice and in vitro experiments of uPA-treated HEK-293 cells. We evaluated renal injury markers, histological changes, mRNA expression of inflammatory, apoptotic, and autophagy markers, as compared with wild-type animals. Results: the αMUPA mice exhibited less renal injury post-AKI, as was evident by lower SCr, BUN, and renal NGAL and KIM-1 along attenuated adverse histological alterations. Notably, the αMUPA mice exhibited decreased levels pro-inflammatory, fibrotic, apoptotic, and autophagy markers like TGF-β, IL-6, STAT3, IKB, MAPK, Caspase-3, and LC3. By contrast, ACE-2, p-eNOS, and PGC1α were higher in the kidneys of the αMUPA mice. In vitro results of the uPA-treated HEK-293 cells mirrored the in vivo findings. Conclusions: These results indicate that uPA modulates key pathways involved in AKI, offering potential therapeutic targets for mitigating renal damage.

The aim of this study was two-fold: (i) to examine the relationships between force outputs measured in the isometric horizontal push test (IHPT) and athletic performances; (ii) to compare IHPT outputs between football players and recreationally active controls. Thirty-two male subjects (football players, n = 16; university students, n = 16) completed the IHPT, countermovement jump (CMJ), standing long jump (SLJ), 5 m, 10 m and 20 m sprint tests, randomly across two testing sessions. Multivariate linear regression analysis was used to examine the relationships between IHPT outputs and athletic performances by accounting for the subjects’ athletic background. An independent sample t-test was used to compare the IHPT outputs between groups. Moderate to very strong linear relationships (r2 range: 0.16–0.56) were found between the IHPT and all athletic performances (all p

We explored the influence of habitat structure on bird density and species richness in the poorly known bird communities in the steppes of Eastern Morocco, along a 200 km long N–S gradient of increasing aridity. The birds were surveyed, and habitat structure was measured in 44 transects regularly distributed along the gradient and during the winter and spring seasons in two consecutive years. After applying a principal component analysis (PCA), five axes were identified, including one related to the latitude–altitude–soil-type gradient and another describing the development of herbaceous vegetation. Generalized linear models were used to explore the relations between bird density and species richness with PCA axes in each season, considering both the entire community and groups of granivorous, insectivorous, and mixed-diet species. More than 90% of the birds were year-round residents, with larks dominating the community in both seasons. We conclude that a distinct multifactorial response can be identified for each functional group of species. In the winter, the community is mainly affected by the structure of the habitat, while aridity (and its assumed relation to primary production) is less influential. In the spring, habitat structure continues to have the greatest explanatory power, but location along the aridity gradient becomes more relevant. These findings reveal the interaction of the negative effects of climatic and anthropogenic changes in the habitat available to these bird communities, with a greater impact expected on birds with diets that include seeds, as well as a general shift of optimal breeding conditions toward more northerly latitudes.

Abstract Carapace width-carapace length (CW/CLRs), carapace width-weight (CW/WRs), and carapace length-weight (CL/WR) relationships are presented for the portunid species Callinectes bocourti A. Milne Edwards, 1879, Callinectes danae Smith, 1869, and Callinectes ornatus Ordway, 1863 from the northern coast of Brazil. A total of 85 crabs were collected between May and October 2015 in intertidal zones of the Caeté, Pirabas and Salinas estuaries. All species had strong relationships between measurements with a coefficient of determination (r 2) ranging between 0.97 and 0.99. The variable most strongly related to CW was CL with 99 % of the variability provided by C. ornatus. These relationships are a useful tool for field ecologists to estimate length and/or mass of portunid species, and it is not necessary to euthanize the organism to collect body measurements. Additionally, this study describes the first reference of CW/CLRs and CL/WRs for these portunid species in the region. The Callinectes species in Amazon estuaries are still poorly studied, and there is a need for monitoring their stocks due to its importance for artisanal fishery. The present data are important for supplying biological information required for an adequate management of this fishery.

Monitoring temporal dynamics in genetic diversity is of great importance for conservation, especially for threatened species that are suffering a rapid population decline and increased fragmentation. Here, we investigate temporal variation in genetic diversity, structure, and gene flow in the Dupont’s lark (Chersophilus duponti) across most of its range. This species shows increasing levels of population fragmentation, substantial population declines, and severe range contraction, so temporal losses of genetic diversity, increasing differentiation, and decreasing gene flow are expected when comparing present day data with previous situations. To address this, we resampled sites (nine regions in two countries) after 12–15 years (five-to-seven generations) and assessed changes in genetic parameters using 11 microsatellite markers. We found no substantial loss in genetic diversity over time at the species level, but we detected considerable variation among regions in the amount of allelic diversity and heterozygosity lost over time. Temporal variation in allele frequencies (common, rare, and private alleles), and changes in genetic differentiation and gene flow over time suggest a major role of connectivity for the stability of the overall metapopulation. Our results agree with the hypothesis that connectivity rescues genetic diversity via immigration and gene flow. However, evidence of recent genetic bottleneck and the substantial changes detected in some regions are clear signs of genetic erosion and may be signalling a rapid decline of the populations. Urgent actions must be carried out to stop and reverse human impacts on this threatened lark and its habitat.

Summary: Primary tauopathies are characterized neuropathologically by inclusions containing abnormal forms of the microtubule-associated protein tau (MAPT) and clinically by diverse neuropsychiatric, cognitive, and motor impairments. Autosomal dominant mutations in the MAPT gene cause heterogeneous forms of frontotemporal lobar degeneration with tauopathy (FTLD-Tau). Common and rare variants in the MAPT gene increase the risk for sporadic FTLD-Tau, including progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD). We generated a collection of fibroblasts from 140 MAPT mutation/risk variant carriers, PSP, CBD, and cognitively normal controls; 31 induced pluripotent stem cell (iPSC) lines from MAPT mutation carriers, non-carrier family members, and autopsy-confirmed PSP patients; 33 genome engineered iPSCs that were corrected or mutagenized; and forebrain neural progenitor cells (NPCs). Here, we present a resource of fibroblasts, iPSCs, and NPCs with comprehensive clinical histories that can be accessed by the scientific community for disease modeling and development of novel therapeutics for tauopathies. : In this article, Karch, Temple and colleagues describe a resource of fibroblasts, patient-derived induced pluripotent stem cells, and genome engineered stem cells with comprehensive clinical histories that can be accessed by the scientific community for disease modeling and development of novel therapeutics for primary tauopathies. Keywords: tau, MAPT, fibroblasts, induced pluripotent stem cells, neural progenitor cells, tauopathy, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration, CRISPR/Cas9

Abstract The family Pseudorhombilidae Alcock, 1900 is widely distributed in the tropical western Atlantic, occurring from shallow waters to the continental slope, inhabiting sandy, gravelly, or muddy substrates. Despite the broad distribution of this family in the Americas, the species Pseudorhombila octodentata Rathbun, 1906 and Pseudorhombila quadridentata (Latreille, 1828) have been observed only in a few localities, with disjunct records, and Robertsella meridionalis Tavares and Gouvêa, 2013 has been recorded only in one type locality (Santa Catarina, Brazil). The present work reports new occurrences of P. octodentata from the state of Sergipe and P. quadridentata from the state of Amapá, and the second occurrence of R. meridionalis from the state of Rio Grande do Norte, Brazil. These new records are an important contribution to the understanding of distributional patterns of pseudorhombilid crab species in Brazil and to the increase in biodiversity data of crustaceans from the North and Northeast regions of Brazil.

Background The aim of this study was to investigate if choice over resistance training exercise order affects motor performance and psychological outcomes among elite youth hockey players. Methods Seventeen elite hockey players (male, n = 14; female, n = 3, age: 15.1 ± 1.1 years) participated in this study. In the first session, individual optimum power loads were calculated in the back squat, jump squat, bench press and bench throw exercises. Then, in four counterbalanced sessions, participants completed three sets of six repetitions in the same exercises loaded with their optimum power loads. In two sessions, athletes used a self-selected order of exercises, while in other two sessions the order was predetermined. Power outputs were estimated with a linear position transducer. Fatigue and enjoyment were measured during and after the sessions using standardized questionnaires. Repeated measures analyses of variance and a paired-sample t-test were used to compare the effects between conditions. Results We observed trivial to small differences between conditions in power outputs (p ≥ 0.07; ES ≤ 0.21), fatigue (p ≥ 0.42; ES ≤ 0.33) and enjoyment (p = 0.72; ES = 0.05). Conclusion Given the comparable effects between approaches, both can be used when coaching youth athletes. Self-selecting the order of exercises based on preferences is a feasible and practical coaching option when working with youth athletes.

In general, a mathematical model that contains many linear/nonlinear differential equations, describing a phenomenon, does not have an explicit hierarchy of system variables. That is, the identification of the fast variables and the slow variables of the system is not explicitly clear. The decomposition of a system into fast and slow subsystems is usually based on intuitive ideas and knowledge of the mathematical model being investigated. In this study, we apply the singular perturbed vector field (SPVF) method to the COVID-19 mathematical model of to expose the hierarchy of the model. This decomposition enables us to rewrite the model in new coordinates in the form of fast and slow subsystems and, hence, to investigate only the fast subsystem with different asymptotic methods. In addition, this decomposition enables us to investigate the stability analysis of the model, which is important in case of COVID-19. We found the stable equilibrium points of the mathematical model and compared the results of the model with those reported by the Chinese authorities and found a fit of approximately 96 percent.

Background While reaching task-failure in resistance-exercises is a topic that attracts scientific and applied interest, the underlying perceived reasons leading to task-failure remain underexplored. Here, we examined the reasons subjects attribute to task-failure as they performed resistance-exercises using different loads. Methods Twenty-two resistance-trained subjects (11-females) completed one Repetition-Maximum (RM) tests in the barbell squat and bench-press. Then, in the next two counterbalanced sessions, subjects performed two sets to task-failure in both exercises, using either 70% or 83% of 1RM. Approximately 30 seconds after set-completion, subjects verbally reported the reasons they perceived to have caused them to reach task-failure. Their answers were recorded, transcribed, and thematically analyzed. The differences between the frequencies of the identified categories were then tested using a mixed logistic regression model. Results The most commonly reported reason was muscle fatigue (54%, p

To use machine vision technology in visual quality control of cereal seeds, sufficient knowledge is necessary. In this work, the capability of machine visual systems, equipped with industrial digital cameras for the identification and classification of seven-grain groups in wheat seed samples, was studied. Two statistical models and three support vector machines were employed in this study. Through image processing of 21,000 single grains, the shape, colour, and textural features of each grain were determined. Ninety-one features were ranked through the ReliefF method. The shape features were the most prominent, followed by the textural and colour features. Among the five models tested, the highest classification accuracy was obtained using quadratic support vector machine (QSVM) and the first 35 features. In the test run of this model with independent data, the classification accuracy for sound white wheat, small white wheat, broken white wheat, shrunken white wheat, red wheat, barley and rye were, respectively, 98.7, 98, 99.3, 90.7, 99, 100, and 97.3%, with an overall average accuracy of 97.6%. In the context of this study, the machine vision system—comprising an industrial digital camera and quadratic support vector machine or non-linear discriminate analysis method—was identified as a valuable system in the investigation of the visual qualities of wheat seeds.

Accurately determining the nutritional status of plants can prevent many diseases caused by fertilizer disorders. Leaf analysis is one of the most used methods for this purpose. However, in order to get a more accurate result, disorders must be identified before symptoms appear. Therefore, this study aims to identify leaves with excessive nitrogen using one-dimensional convolutional neural networks (1D-CNN) on a dataset of spectral data using the Keras library. Seeds of cucumber were planted in several pots and, after growing the plants, they were divided into different classes of control (without excess nitrogen), N30% (excess application of nitrogen fertilizer by 30%), N60% (60% overdose), and N90% (90% overdose). Hyperspectral data of the samples in the 400–1100 nm range were captured using a hyperspectral camera. The actual amount of nitrogen for each leaf was measured using the Kjeldahl method. Since there were statistically significant differences between the classes, an individual prediction model was designed for each class based on the 1D-CNN algorithm. The main innovation of the present research resides in the application of separate prediction models for each class, and the design of the proposed 1D-CNN regression model. The results showed that the coefficient of determination and the mean squared error for the classes N30%, N60% and N90% were 0.962, 0.0005; 0.968, 0.0003; and 0.967, 0.0007, respectively. Therefore, the proposed method can be effectively used to detect over-application of nitrogen fertilizers in plants.

Cross-presentation initiates immune responses against tumors and viral infections by presenting extracellular antigen on MHC I to activate CD8+ T cell-mediated cytotoxicity. In vitro studies in dendritic cells (DCs) established SNARE protein SEC22B as a specific regulator of cross-presentation. However, the in vivo contribution of SEC22B to cross-presentation has not been tested. To address this, we generated DC-specific Sec22b knockout (CD11c-Cre Sec22bfl/fl) mice. Contrary to the paradigm, SEC22B-deficient DCs efficiently cross-present both in vivo and in vitro. Although in vitro small hairpin RNA (shRNA)-mediated Sec22b silencing in bone-marrow-derived dendritic cells (BMDCs) reduced cross-presentation, treatment of SEC22B-deficient BMDCs with the same shRNA produced a similar defect, suggesting the Sec22b shRNA modulates cross-presentation through off-target effects. RNA sequencing of Sec22b shRNA-treated SEC22B-deficient BMDCs demonstrated several changes in the transcriptome. Our data demonstrate that contrary to the accepted model, SEC22B is not necessary for cross-presentation, cautioning against extrapolating phenotypes from knockdown studies alone.

Today, as fossil fuels are depleted, renewable energy must be used to meet the needs of human beings. One of the renewable energy sources is undoubtedly the solar–geothermal power plant. In this paper, the conventional and advanced, exergo-environmental and exergo-economic analysis of a geothermal–solar hybrid power plant (SGHPP) based on an organic Rankin cycle (ORC) cycle is investigated. In this regard, at first, a conventional analysis was conducted on a standalone geothermal cycle (first mode), as well as a hybrid solar–geothermal cycle (second mode). The results of exergy destruction for simulating the standalone geothermal cycle showed that the ORC turbine with 1050 kW had the highest exergy destruction that was 38% of the total share of destruction. Then, the ORC condenser with 26% of the total share of exergy destruction was in second place. In the hybrid geothermal–solar cycle, the solar panel had the highest environmental impact and about 56% of the total share of exergy destruction. The ORC turbine had about 9% of all exergy destruction. The results of the advanced analysis of exergy in the standalone geothermal cycle showed that the avoidable exergy destruction of the condenser was the highest. In the hybrid geothermal–solar cycle, the solar panel, steam economizer and steam evaporator were ranked first to third from an avoidable exergy destruction perspective. The avoidable exergo-economic destruction of the evaporator and pump were higher than the other components. The hybrid geothermal–solar cycle, steam economizer, solar pane and steam evaporator were ranked first to third, respectively, and they could be modified. The avoidable exergo-environmental destruction of the ORC turbine and the ORC pump were the highest, respectively. In the hybrid geothermal–solar cycle, steam economizers, solar panel and steam evaporators had the highest avoidable exergy destruction, respectively. For the standalone geothermal cycle, the total endogenous exergy destruction and exogenous exergy destruction was 83.61% and 16.39%. Moreover, from an exergo-economic perspective, 89% of the total destruction rate was endogenous and 11% was exogenous. From an exergo-environmental perspective, 88.73% of the destruction rate was endogenous and 11.27% was exogenous. For the hybrid geothermal–solar cycle, the total endogenous and exogenous exergy destruction was 75.08% and 24.92%, respectively. Moreover, 81.82% of the exergo-economic destruction rate was endogenous and 18.82% was exogenous. From an exergo-environmental perspective, 81.19% of the exergy destruction was endogenous and 18.81% was exogenous.

Music has been shown to reduce rating of perceived exertion, increase exercise enjoyment and enhance exercise performance, mainly in low-moderate intensity exercises. However, the effects of music are less conclusive with high-intensity activities. The purpose of this with-participant design study was to compare the effects of high tempo music (130 bpm) to a no-music condition during repeated high intensity cycling bouts (80% of peak power output (PPO)) on the following measures: time to exercise end-point, rating of perceived exertion (RPE), heart rate (HR), breathing frequency, ventilatory kinetics and blood lactate (BL). Under the music condition, participants exercised 10.7% longer (p = 0.035; Effect size (ES) = 0.28) (increase of 1 min) and had higher HR (4%; p = 0.043; ES = 0.25), breathing frequency (11.6%; p < 0.001; ES = 0.57), and RER (7% at TTF; p = 0.021; ES = 1.1) during exercise, as measured at the exercise end-point. Trivial differences were observed between conditions in RPE and other ventilatory kinetics during exercise. Interestingly, 5 min post-exercise termination, HR recovery was 13.0% faster following the music condition (p < 0.05) despite that music was not played during this period. These results strengthen the notion that music can alter the association between central motor drive, central cardiovascular command and perceived exertion, and contribute to prolonged exercise durations at higher intensities along with a quicken HR recovery.

Nonmembrane-bound organelles that behave like liquid droplets are widespread among eukaryotic cells. Their dysregulation appears to be a critical step in several neurodegenerative conditions. Here, we report that tau protein, the primary constituent of Alzheimer neurofibrillary tangles, can form liquid droplets and therefore has the necessary biophysical properties to undergo liquid-liquid phase separation (LLPS) in cells. Consonant with the factors that induce LLPS, tau is an intrinsically disordered protein that complexes with RNA to form droplets. Uniquely, the pool of RNAs to which tau binds in living cells are tRNAs. This phase state of tau is held in an approximately 1:1 charge balance across the protein and the nucleic acid constituents, and can thus be maximal at different RNA:tau mass ratios, depending on the biopolymer constituents involved. This feature is characteristic of complex coacervation. We furthermore show that the LLPS process is directly and sensitively tuned by salt concentration and temperature, implying it is modulated by both electrostatic interactions between the involved protein and nucleic acid constituents, as well as net changes in entropy. Despite the high protein concentration within the complex coacervate phase, tau is locally freely tumbling and capable of diffusing through the droplet interior. In fact, tau in the condensed phase state does not reveal any immediate changes in local protein packing, local conformations and local protein dynamics from that of tau in the dilute solution state. In contrast, the population of aggregation-prone tau as induced by the complexation with heparin is accompanied by large changes in local tau conformations and irreversible aggregation. However, prolonged residency within the droplet state eventually results in the emergence of detectable β-sheet structures according to thioflavin-T assay. These findings suggest that the droplet state can incubate tau and predispose the protein toward the formation of insoluble fibrils.

In this study, the spatial distribution of PM2.5 air pollution in Mexico City from 37 personal exposures was modeled. Meteorological, demographic, geographic, and social data were also included. Geographic information systems (GIS), spatial analysis, and Land-Use Regression (LUR) were used to generate the final predictive model and the spatial distribution map which revealed two areas with very high concentrations (up to 109.3 µg/m3) and two more with lower concentrations (between 72 to 86.5 µg/m3) (p < 0.05). These results illustrate an overview trend of PM2.5 in relation to human activity during the studied periods in Mexico City and show a general approach to understanding the spatial variability of PM2.5.

Community-based natural resource management (CBNRM) is a concept critical to managing social-ecological systems but whose implementation needs strengthening. Scenario planning is one approach that may offer benefits relevant to CBNRM but whose potential is not yet well understood. Therefore, we designed, trialed, and evaluated a scenario-planning method intended to support CBNRM in three cases, located in Colombia, Mexico, and Argentina. Implementing scenario planning was judged as worthwhile in all three cases, although aspects of it were challenging to facilitate. The benefits generated were relevant to strengthening CBNRM: encouraging the participation of local people and using their knowledge, enhanced consideration of and adaptation to future change, and supporting the development of systems thinking. Tracing exactly when and how these benefits arose was challenging, but two elements of the method seemed particularly useful. First, using a systematic approach to discuss how drivers of change may affect local social-ecological systems helped to foster systems thinking and identify connections between issues. Second, explicitly focusing on how to use and respond to scenarios helped identify specific practical activities, or "response options," that would support CBNRM despite the pressures of future change. Discussions about response options also highlighted the need for support by other actors, e.g., policy groups: this raised the question of when and how other actors and other sources of knowledge should be involved in scenario planning, so as to encourage their buy-in to actions identified by the process. We suggest that other CBNRM initiatives may benefit from adapting and applying scenario planning. However, these initiatives should be carefully monitored because further research is required to understand how and when scenario-planning methods may produce benefits, as well as their strengths and weaknesses versus other methods.