Defining the Effects of PKC Modulator HIV Latency-Reversing Agents on Natural Killer Cells.

Background: Latency reversing agents (LRAs) such as protein kinase C (PKC) modulators can reduce rebound-competent HIV reservoirs in small animal models. Furthermore, administration of natural killer (NK) cells following LRA treatment improves this reservoir reduction. It is currently unknown why the combination of a PKC modulator and NK cells is so potent and whether exposure to PKC modulators may augment NK cell function in some way. Methods: Primary human NK cells were treated with PKC modulators (bryostatin-1, prostratin, or the designed, synthetic bryostatin-1 analog SUW133), and evaluated by examining expression of activation markers by flow cytometry, analyzing transcriptomic profiles by RNA sequencing, measuring cytotoxicity by co-culturing with K562 cells, assessing cytokine production by Luminex assay, and examining the ability of cytokines and secreted factors to independently reverse HIV latency by co-culturing with Jurkat-Latency (J-Lat) cells. Results: PKC modulators increased expression of proteins involved in NK cell activation. Transcriptomic profiles from PKC-treated NK cells displayed signatures of cellular activation and enrichment of genes associated with the NFκB pathway. NK cell cytotoxicity was unaffected by prostratin but significantly decreased by bryostatin-1 and SUW133. Cytokines from PKC-stimulated NK cells did not induce latency reversal in J-Lat cell lines. Conclusions: Although PKC modulators have some significant effects on NK cells, their contribution in "kick and kill" strategies is likely due to upregulating HIV expression in CD4+ T cells, not directly enhancing the effector functions of NK cells. This suggests that PKC modulators are primarily augmenting the "kick" rather than the "kill" arm of this HIV cure approach.

Cutaneous Mucormycosis in Buffalos in the Brazilian Amazon Biome.

This is the first description of cutaneous mucormycosis in buffalo in the Brazilian Amazon biome. All buffalo showed apathy, inappetence, weight loss, reluctance to move, and prolonged sternal decubitus. Of the four affected animals, two died 15 and 30 days after the appearance of clinical signs. In the initial phase, the skin lesions were rounded areas with dry central regions, sensitive to palpation, with protruding edges and diameters ranging from 8 cm to 15 cm. These areas of necrosis were isolated or coalescing and present mainly on the limbs and sides. In an advanced stage of the disease, there was detachment of the skin from the necrotic areas with extensive wound formation, which sometimes exposed the subcutaneous tissue. The histopathology of the skin showed a multifocal inflammatory infiltrate composed of intact and degenerated eosinophils surrounded by epithelioid macrophages. At the center of these areas was a focally extensive area of epidermal ulceration characterized by intact and degenerated neutrophils, the necrosis of epithelial cells, and the accumulation of fibrin and erythrocytes. The mycological culture was positive for Rhizopus sp. The diagnosis of cutaneous dermatitis caused by Rhizopus sp. was based on clinical signs, macroscopic and histopathological findings, and the identification of the fungus by mycological and molecular techniques.

Complement receptor 3 is required for maximum in vitro trogocytic killing of the parasite Trichomonas vaginalis by human neutrophil-like cells.

Trichomonas vaginalis (Tv) is a parasite that causes trichomoniasis, a prevalent sexually-transmitted infection. Neutrophils are found at the site of infection, and can rapidly kill the parasite in vitro, using trogocytosis. However, the specific molecular players in neutrophil killing of Tv are unknown. Here, we show that complement proteins play a role in Tv killing by human neutrophil-like cells (NLCs). Using CRISPR/Cas9, we generated NLCs deficient in each of three complement receptors (CRs) known to be expressed on human neutrophils: CR1, CR3, and CR4. Using in vitro trogocytosis assays, we found that CR3, but not CR1 or CR4 is required for maximum trogocytosis of the parasite by NLCs, with NLCs lacking CR3 demonstrating ~40% reduction in trogocytosis, on average. We also observed a reduction in NLC killing of Tv in CR3 knockout, but not CR1 or CR4 knockout NLCs. On average, NLCs lacking CR3 had ~50% reduction in killing activity. We also used a parallel approach of pre-incubating NLCs with blocking antibodies against CR3, which similarly reduced NLC killing of parasites. These data support a model in which Tv is opsonized by the complement protein iC3b, and bound by neutrophil CR3 receptor, to facilitate trogocytic killing of the parasite.

A new oxidative pathway of nitric oxide production from oximes in plants.

Nitric oxide (NO) is an essential reactive oxygen species and a signal molecule in plants. Although several studies have proposed the occurrence of oxidative NO production, only reductive routes for NO production, such as the nitrate (NO-3) -upper-reductase pathway, have been evidenced to date in land plants. However, plants grown axenically with ammonium as the sole source of nitrogen exhibit contents of nitrite and NO3-, evidencing the existence of a metabolic pathway for oxidative production of NO. We hypothesized that oximes, such as indole-3-acetaldoxime (IAOx), a precursor to indole-3-acetic acid, are intermediate oxidation products in NO synthesis. We detected the production of NO from IAOx and other oximes catalyzed by peroxidase (POD) enzyme using both 4-amino-5-methylamino-2',7'-difluorescein fluorescence and chemiluminescence. Flavins stimulated the reaction, while superoxide dismutase inhibited it. Interestingly, mouse NO synthase can also use IAOx to produce NO at a lower rate than POD. We provided a full mechanism for POD-dependent NO production from IAOx consistent with the experimental data and supported by density functional theory calculations. We showed that the addition of IAOx to extracts from Medicago truncatula increased the in vitro production of NO, while in vivo supplementation of IAOx and other oximes increased the number of lateral roots, as shown for NO donors, and a more than 10-fold increase in IAOx dehydratase expression. Furthermore, we found that in vivo supplementation of IAOx increased NO production in Arabidopsis thaliana wild-type plants, while prx33-34 mutant plants, defective in POD33-34, had reduced production. Our data show that the release of NO by IAOx, as well as its auxinic effect, explain the superroot phenotype. Collectively, our study reveals that plants produce NO utilizing diverse molecules such as oximes, POD, and flavins, which are widely distributed in the plant kingdom, thus introducing a long-awaited oxidative pathway to NO production in plants. This knowledge has essential implications for understanding signaling in biological systems.

Control technologies to prevent aerosol-based disease transmission in animal agriculture production settings: a review of established and emerging approaches.

Transmission of infectious agents via aerosols is an ever-present concern in animal agriculture production settings, as the aerosol route to disease transmission can lead to difficult-to-control and costly diseases, such as porcine respiratory and reproductive syndrome virus and influenza A virus. It is increasingly necessary to implement control technologies to mitigate aerosol-based disease transmission. Here, we review currently utilized and prospective future aerosol control technologies to collect and potentially inactivate pathogens in aerosols, with an emphasis on technologies that can be incorporated into mechanically driven (forced air) ventilation systems to prevent aerosol-based disease spread from facility to facility. Broadly, we find that control technologies can be grouped into three categories: (1) currently implemented technologies; (2) scaled technologies used in industrial and medical settings; and (3) emerging technologies. Category (1) solely consists of fibrous filter media, which have been demonstrated to reduce the spread of PRRSV between swine production facilities. We review the mechanisms by which filters function and are rated (minimum efficiency reporting values). Category (2) consists of electrostatic precipitators (ESPs), used industrially to collect aerosol particles in higher flow rate systems, and ultraviolet C (UV-C) systems, used in medical settings to inactivate pathogens. Finally, category (3) consists of a variety of technologies, including ionization-based systems, microwaves, and those generating reactive oxygen species, often with the goal of pathogen inactivation in aerosols. As such technologies are typically first tested through varied means at the laboratory scale, we additionally review control technology testing techniques at various stages of development, from laboratory studies to field demonstration, and in doing so, suggest uniform testing and report standards are needed. Testing standards should consider the cost-benefit of implementing the technologies applicable to the livestock species of interest. Finally, we examine economic models for implementing aerosol control technologies, defining the collected infectious particles per unit energy demand.

HIV establishes an early foothold.

The weeks following HIV acquisition are a critical time when the virus causes significant immunological damage and establishes long-term latent reservoirs. A recent study in Immunity by Gantner et al. uses single-cell analysis to explore these key early infection events, providing insights into early HIV pathogenesis and reservoir formation.

Indole-3-acetaldoxime delays root iron-deficiency responses and modify auxin homeostasis in Medicago truncatula.

Iron (Fe) is an essential plant micronutrient, being a major limiting growth factor in calcareous soils. To increase Fe uptake, plants induce lateral roots growth, the expression of a Fe(III)-chelate reductase (FCR), a Fe(II)-transporter and a H+-ATPase and the secretion of flavins. Furthermore, auxin hormone family is involved in the Fe-deficiency responses but the action mechanism remains elusive. In this work, we evaluated the effect of the auxin-precursor indole-3-acetaldoxime (IAOx) on hydroponically grown Medicago truncatula plants under different Fe conditions. Upon 4-days of Fe starvation, the pH of the nutrient solution decreased, while both the FCR activity and the presence of flavins increased. Exogenous IAOx increased lateral roots growth contributing to superroot phenotype, decreased chlorosis, and delayed up to 3-days the pH-decrease, the FCR-activity increase, and the presence of flavins, compared to Fe-deficient plants. Gene expression levels were in concordance with the physiological responses. RESULTS: showed that IAOx was immediately transformed to IAN in roots and shoots to maintain auxin homeostasis. IAOx plays an active role in iron homeostasis delaying symptoms and responses in Fe-deficient plants. We may speculate that IAOx or its derivatives remobilize Fe from root cells to alleviate Fe-deficiency. Overall, these results point out that the IAOx-derived phenotype may have advantages to overcome nutritional stresses.

Secreted factors induced by PKC modulators do not indirectly cause HIV latency reversal.

HIV can establish a long-lived latent infection in cells harboring integrated non-expressing proviruses. Latency reversing agents (LRAs), including protein kinase C (PKC) modulators, can induce expression of latent HIV, thereby reducing the latent reservoir in animal models. However, PKC modulators such as bryostatin-1 also cause cytokine upregulation in peripheral blood mononuclear cells (PBMCs), including cytokines that might independently reverse HIV latency. To determine whether cytokines induced by PKC modulators contribute to latency reversal, primary human PBMCs were treated with bryostatin-1 or the bryostatin analog SUW133, a superior LRA, and supernatant was collected. As anticipated, LRA-treated cell supernatant contained increased levels of cytokines compared to untreated cell supernatant. However, exposure of latently-infected cells with this supernatant did not result in latency reactivation. These results indicate that PKC modulators do not have significant indirect effects on HIV latency reversal in vitro and thus are targeted in their latency reversing ability.

A Study of the Interface of Gold Nanoparticles Conjugated to Cowpea Fe-Superoxide Dismutase.

The iron superoxide dismutase (FeSOD) is a first barrier to defend photosynthetic organisms from superoxide radicals. Although it is broadly present in plants and bacteria, FeSODs are absent in animals. They belong to the same phylogenic family as Mn-containing SODs, which are also highly efficient at detoxifying superoxide radicals. In addition, SODs can react with peroxynitrite, and FeSOD enzyme has already been used to evaluate the anti-nitrative capacity of plant antioxidants. Gold nanoparticles (AuNPs) have been shown to significantly improve the functionality and the efficiency of ligands, providing they are properly assembled. In this work, the characteristics of the recombinant cowpea (Vigna unguiculata) FeSOD (rVuFeSOD) immobilized onto AuNPs were investigated as a function of (1) NP surface chemistry and (2) biofunctionalization methods, either physical adsorption or covalent bonding. The NP surface chemistry was studied by varying the concentration of the ligand molecule 11-mercaptoundecanoic acid (MUA) on the NP surface. The coverage and activity of the protein on AuNPs was determined and correlated to the surface chemistry and the two biofunctionalization methods. rVuFeSOD-AuNPs conjugate stability was monitored through absorption measurements, agarose gel electrophoresis and DLS, enzymatic activity by a colorimetric assay and by in-gel activity assay, and coverage was measured by colorimetric assay. When using physical adsorption, the NP is the most perturbing agent for the activity of the enzyme. In contrast, only the NP coverage was affected by MUA ligand concentration. However, during covalent attachment, both the NP and the concentration of MUA on the surface influenced the enzyme activity, while the coverage of the NP remained constant. The results evidence the importance of the biomolecule and AuNP interaction for the functionality of the hybrid. These strategies can be used to develop electrochemical biosensors for O2•- and for peroxynitrite in biomedical applications.

Contribution of Sex Differences to HIV Immunology, Pathogenesis, and Cure Approaches.

Approximately 38 million people were living with human immunodeficiency virus (HIV) in 2020 and 53% of those infected were female. A variety of virological and immunological sex-associated differences (sexual dimorphism) in HIV infection have been recognized in males versus females. Social, behavioral, and societal influences play an important role in how the HIV pandemic has affected men and women differently. However, biological factors including anatomical, physiologic, hormonal, and genetic differences in sex chromosomes can each contribute to the distinct characteristics of HIV infection observed in males versus females. One striking example of this is the tendency for women to have lower HIV plasma viral loads than their male counterparts early in infection, though both progress to AIDS at similar rates. Sex differences in acquisition of HIV, innate and adaptive anti-HIV immune responses, efficacy/suitability of specific antiretroviral drugs, and viral pathogenesis have all been identified. Sex differences also have the potential to affect viral persistence, latency, and cure approaches. In this brief review, we summarize the major biological male/female sex differences in HIV infection and their importance to viral acquisition, pathogenesis, treatment, and cure efforts.

Body Composition and Nutrients Dietary Intake Changes during COVID-19 Lockdown in Spanish Healthy Postmenopausal Women.

(1) Background: During the COVID-19 lockdown, high rates of physical inactivity and dietary imbalances were reported in both adults and adolescents. Physical separation and isolation not only have a significant impact on the performance of physical activity but also affect people's lives, particularly their dietary habits. In the present study, we aimed to examine whether or not bioelectrical impedance-derived body composition parameters and dietary habits were affected during the pandemic-associated lockdown in postmenopausal Spanish women. (2) Methods: Sixty-six women participated in the study (58.7 ± 5.4 years) before (between July−October 2019) and after (August−October 2020) the lockdown, which occurred as a consequence of the COVID-19 pandemic in Spain. Body composition parameters were measured by bioelectrical impedance analysis while dietary intake of proteins, fat, carbohydrates, and energy was measured by a food frequency questionnaire. (3) Results Regarding body composition, no differences were observed in fat mass in % (mean increase 0.05 (2.74); p = 0.567), fat mass in kg (mean increase −0.07 (4.137); p = 0.356) or lean mass in kg (mean increase 0.20 (1.424); p = 0.636). Similarly, no statistically significant differences were observed between the two study periods for any of the nutrients studied, nor for energy intake (p > 0.05 in all cases). (4) Conclusions: After comprehensively assessing body composition and dietary intake of protein, fat, carbohydrates, and energy before and after COVID-19 lockdown in healthy adult women in Spain no changes in the parameters studied were observed during the period analyzed in the women examined.

"Por las razones adecuadas": producción y competencias científicas en la investigación en terapia ocupacional / "For the right reasons": scientific production and competencies in occupational therapy research

La investigación de calidad es una de las piedras angulares sobre la que se sustenta la práctica basada en la evidencia. De esta forma, tanto leer como producir ciencia nos pone en el camino para realizar la mejor toma de decisiones posible sobre los procedimientos a aplicar junto con los usuarios. En estos últimos años se han evidenciado las limitaciones y barreras a las que se enfrentan los y las terapeutas ocupacionales a la hora de implementar una terapia ocupacional basada en la evidencia de calidad en su práctica diaria: mayor necesidad de formación metodológica, en bioestadística y/o dificultad para analizar la literatura disponible. Además, cada vez se tiene una mayor conciencia de la existencia de una brecha entre el ejercicio profesional y la investigación, lo que esperamos invite a reducir dicha brecha para poder ofrecer a la sociedad una práctica basada en las mejores evidencias disponibles.

High quality research is one of the foundations on which evidence-based occupational therapy is sustained. In this way, both reading and producing science allows us to take the best decisions about the most appropriate approaches to apply with our users. In recent years, the limitations and barriers faced by occupational therapists implementing an evidence-based occupational therapy in their daily practice have become obvious: a greater need for both methodological and biostatistics training, or the difficulty in analyzing the available literature. But there is a growing awareness of the existence of a gap between professional practice and research, which we hope will help to reduce this gap to offer society a practice based on the best available evidence.

The importance of the urea cycle and its relationships to polyamine metabolism during ammonium stress in Medicago truncatula.

The ornithine-urea cycle (urea cycle) makes a significant contribution to the metabolic responses of lower photosynthetic eukaryotes to episodes of high nitrogen availability. In this study, we compared the role of the plant urea cycle and its relationships to polyamine metabolism in ammonium-fed and nitrate-fed Medicago truncatula plants. High ammonium resulted in the accumulation of ammonium and pathway intermediates, particularly glutamine, arginine, ornithine, and putrescine. Arginine decarboxylase activity was decreased in roots, suggesting that the ornithine decarboxylase-dependent production of putrescine was important in situations of ammonium stress. The activity of copper amine oxidase, which releases ammonium from putrescine, was significantly decreased in both shoots and roots. In addition, physiological concentrations of ammonium inhibited copper amine oxidase activity in in vitro assays, supporting the conclusion that high ammonium accumulation favors putrescine synthesis. Moreover, early supplementation of plants with putrescine avoided ammonium toxicity. The levels of transcripts encoding urea-cycle-related proteins were increased and transcripts involved in polyamine catabolism were decreased under high ammonium concentrations. We conclude that the urea cycle and associated polyamine metabolism function as important protective mechanisms limiting ammonium toxicity in M. truncatula. These findings demonstrate the relevance of the urea cycle to polyamine metabolism in higher plants.

Tryptophan Levels as a Marker of Auxins and Nitric Oxide Signaling.

The aromatic amino acid tryptophan is the main precursor for indole-3-acetic acid (IAA), which involves various parallel routes in plants, with indole-3-acetaldoxime (IAOx) being one of the most common intermediates. Auxin signaling is well known to interact with free radical nitric oxide (NO) to perform a more complex effect, including the regulation of root organogenesis and nitrogen nutrition. To fathom the link between IAA and NO, we use a metabolomic approach to analyze the contents of low-molecular-mass molecules in cultured cells of Arabidopsis thaliana after the application of S-nitrosoglutathione (GSNO), an NO donor or IAOx. We separated the crude extracts of the plant cells through ion-exchange columns, and subsequent fractions were analyzed by gas chromatography-mass spectrometry (GC-MS), thus identifying 26 compounds. A principal component analysis (PCA) was performed on N-metabolism-related compounds, as classified by the Kyoto Encyclopedia of Genes and Genomes (KEGG). The differences observed between controls and treatments are mainly explained by the differences in Trp contents, which are much higher in controls. Thus, the Trp is a shared response in both auxin- and NO-mediated signaling, evidencing some common signaling mechanism to both GSNO and IAOx. The differences in the low-molecular-mass-identified compounds between GSNO- and IAOx-treated cells are mainly explained by their concentrations in benzenepropanoic acid, which is highly associated with IAA levels, and salicylic acid, which is related to glutathione. These results show that the contents in Trp can be a marker for the study of auxin and NO signaling.

Improving the Reliability of Literature Reviews: Detection of Retracted Articles through Academic Search Engines.

Nowadays, a multitude of scientific publications on health science are being developed that require correct bibliographic search in order to avoid the use and inclusion of retracted literature in them. The use of these articles could directly affect the consistency of the scientific studies and could affect clinical practice. The aim of the present study was to evaluate the capacity of the main scientific literature search engines, both general (Gooogle Scholar) and scientific (PubMed, EMBASE, SCOPUS, and Web of Science), used in health sciences in order to check their ability to detect and warn users of retracted articles in the searches carried out. The sample of retracted articles was obtained from RetractionWatch. The results showed that although Google Scholar was the search engine with the highest capacity to retrieve selected articles, it was the least effective, compared with scientific search engines, at providing information on the retraction of articles. The use of different scientific search engines to retrieve as many scientific articles as possible, as well as never using only a generic search engine, is highly recommended. This will reduce the possibility of including retracted articles and will avoid affecting the reliability of the scientific studies carried out.

Relationship between the distribution of vegetation and the environment in the coastal embryo dunes of Jalisco, México.

Background: The poorly developed soils of the embryo dunes imply little capacity for plant support, however, the adaptation mechanisms of plants respond sensitively to environmental variations, even when these variations are small, which results in a set of specialized habitats and flora that are rarely shared with other terrestrial ecosystems. The coastal dunes of the Mexican Pacific remain vaguely studied, this is why this research explored the relationship between environmental properties and the presence of plant species in the embryo dunes of the coast of Jalisco, Mexico. Methods: Twenty-nine sites were sampled, one or two sites per embryo dune, with a random stratified design. Geomorphological and vegetation data were collected at site. Laboratory determinations included soil color, particle size, organic matter, pH, electrical conductivity, magnetite content, and moisture retention. Statistical analysis included correlation analysis to identify relationships between environmental variables; principal component analysis (PCA) and cluster analysis to group dune sites by environmental properties; canonical correspondence analysis (CCA) to determine a possible significant relationship between the presence of plant species and environmental variables; cluster analysis to group dune sites by presence/absence of plant species and correlate both clusters to validate the relationship between them, the salient aspects of this relationship were described and the spatial distribution of the groups was mapped. Results: Eleven plant species were identified, six of them exclusive to the embryo dunes and the rest ubiquitous. The incipient development of these soils is reflected in a low content of organic matter, silt, clay, and moisture retention, with scattered data on granulometry, electrical conductivity, organic matter, and magnetite. Some significant correlations were found between some environmental properties, and the CCA showed a significant relationship between the presence of plant species and environmental variables (p-value of the Monte Carlo test = 0.026). The cluster analysis of dune sites according to environmental variables and the cluster analysis by presence/absence of plant species produces the formation of five groups of sites with significant environmental differences and five groups of sites with significant floristic differences. A significant connection (r = 0.471, p = 0.01) between the two clustering schemes also evidences the meaningful relationship between the presence of plant species and the environmental characteristics of the embryo dunes of Jalisco, Mexico. Differences in habitat preferences were observed among plant species exclusive to the embryo dunes; thus, Abronia maritima, Uniola pittieri, and Pectis arenaria showed a preference for embryo dunes with poor edaphic conditions, in contrast to Okenia hypogaea, Canavalia rosea, and Scaevola plumieri, which were mostly found in embryo dunes with higher fertility.

Nutrition and Women's Bone Health.

Nutrition is a key element that has the potential to reduce bone loss or fracture risk [...].

SCRUTATIOm: cómo detectar literatura retractada incluida en revisiones sistemáticas y metaanálisis usando SCOPUS© y ZOTERO© / SCRUTATIOm: how to detect retracted literature included in systematics reviews and metaanalysis using SCOPUS© and ZOTERO©

Una buena y precisa revisión de la literatura es un elemento clave de las revisiones sistemáticas y de los metaanálisis, pues deben basarse en estudios originales de alta calidad y lógicamente nunca retractados. El objetivo de la presente nota metodológica es proporcionar un sistema rápido, reproducible y fiable de detección de literatura retractada incluida en revisiones sistemáticas y metaanálisis, que permite verificar la consistencia de los resultados y habilita la posibilidad de poder comunicar la potencial presencia de errores a las revistas implicadas mediante un proceso de revisión posterior a la publicación (post peer review). La aplicación sistemática del método SCRUTATIOm permitiría saber si en nuestra área de trabajo se ha publicado un determinado estudio que puede no aportar evidencia consistente al cuerpo del conocimiento. (AU)

A good and accurate literature review is a key element of systematic reviews and meta-analyses and always needs to be based on high quality original studies and logically never retracted. The aim of this methodological note is to provide a fast, reproducible and reliable system for detecting retracted literature included in systematic reviews and meta-analyses, allowing the consistency of the results of these to be verified and enabling the possibility of being able to communicate the potential presence of errors to the journals involved by means of a post-publication or post-peer review process. The SCRUTATIOm method, apply routinely, allows us to know if a certain study has been published in our field of study, which may not provide consistent evidence to the body of knowledge. (AU)

[SCRUTATIOm: how to detect retracted literature included in systematics reviews and metaanalysis using SCOPUS© and ZOTERO©]. / SCRUTATIOm: cómo detectar literatura retractada incluida en revisiones sistemáticas y metaanálisis usando SCOPUS© y ZOTERO©.

A good and accurate literature review is a key element of systematic reviews and meta-analyses and always needs to be based on high quality original studies and logically never retracted. The aim of this methodological note is to provide a fast, reproducible and reliable system for detecting retracted literature included in systematic reviews and meta-analyses, allowing the consistency of the results of these to be verified and enabling the possibility of being able to communicate the potential presence of errors to the journals involved by means of a post-publication or post-peer review process. The SCRUTATIOm method, apply routinely, allows us to know if a certain study has been published in our field of study, which may not provide consistent evidence to the body of knowledge.