Search

Abstract Dengue (DENV) and chikungunya (CHIKV) viruses are among the most preponderant arboviruses. Although primarily transmitted through the bite of Aedes aegypti mosquitoes, Aedes albopictus and Aedes malayensis are competent vectors and have an impact on arbovirus epidemiology. Here, to fill the gap in our understanding of the molecular interactions between secondary vectors and arboviruses, we used transcriptomics to profile the whole-genome responses of A. albopictus to CHIKV and of A. malayensis to CHIKV and DENV at 1 and 4 days post-infection (dpi) in midguts. In A. albopictus, 1793 and 339 genes were significantly regulated by CHIKV at 1 and 4 dpi, respectively. In A. malayensis, 943 and 222 genes upon CHIKV infection, and 74 and 69 genes upon DENV infection were significantly regulated at 1 and 4 dpi, respectively. We reported 81 genes that were consistently differentially regulated in all the CHIKV-infected conditions, identifying a CHIKV-induced signature. We identified expressed immune genes in both mosquito species, using a de novo assembled midgut transcriptome for A. malayensis, and described the immune architectures. We found the JNK pathway activated in all conditions, generalizing its antiviral function to Aedines. Our comprehensive study provides insight into arbovirus transmission by multiple Aedes vectors.

Purpose: Patient-reported outcome measures (PROMs) for persons with aphasia (PWA) are commonly used to measure various outcomes. Persons with aphasia commonly present with language deficits that can likely increase respondent burden. Elements that contribute to respondent burden include readability, design, and formatting methods. The aims of this study were to determine how PROMs for PWA rate in level of respondent burden and on readability levels. Method: Irwin's 2012 review of PROMs for PWA was used for inclusion criteria and additional PROMs developed after 2012 were identified. This method resulted in 14 PROMs, which were rated on proposed respondent burden criteria and also underwent a readability analysis. Results: Six PROMs achieved a 7 or higher with the proposed 10-point respondent burden measure, indicating lower respondent burden. Eight PROMs met the nationally recommended reading level of sixth grade. Conclusions: Results indicated that the several PROMs available for PWA fail to minimize respondent burden.

The definitions and features of "resilience," the process and outcome of successfully responding to adversity, are variable across the disability and rehabilitation literatures, and its influence on aphasia treatment and outcomes is as of yet unknown. This scoping review identified and thematically categorized characterizations and descriptions of resilience and the similarly used terms "adaptation," "adjustment," and "coping" in the aphasia research literature. Frequently noted features of resilience included internal/cognitive emotional responses, such as positive self-concept, attitude/outlook, and psychological function, as well as external/behavioral responses, such as engaging in new activities and opportunities for social connection. These general features align with a definition of resilience recently developed for individuals with chronic impairment. And, although little information about the influence of resilience on treatment engagement and outcomes emerged from the reviewed literature, a number of clinical considerations and research directions are suggested.

Abstract Conformational dynamics plays a critical role for the function of multidomain electron transfer complexes. While crystallographic or NMR approaches allow detailed insight into structures, lower resolution methods like cryo-electron microscopy can provide more information on dynamics. In silico structure modelling using AlphaFold was recently successfully extended to the prediction of protein complexes but its capability to address large conformational changes involved in catalysis remained obscure. We used bacterial CYP102A1 monooxygenase homodimer as a test case to design a competitive modelling approach (CMA) for assessing alternate conformations of multi-domain complexes. Predictions were confronted with published crystallographic and cryo-EM data, evidencing consistencies but also permitting some reinterpretation of experimental data. Structural determinants stabilising the new type of domain connectivity evidenced in this bacterial self-sufficient monooxygenase were analysed by CMA and used for in silico retro-engineering applied to its eukaryotic bi-component counterparts.

The global public health burden exerted by viruses partially stems from viruses’ ability to subdue host cells into creating an environment that promotes their multiplication (i.e., pro-viral). It has been discovered that viruses alter cell physiology by transferring viral material through extracellular vesicles (EVs), which serve as vehicles for intercellular communication. Here, we aim to provide a conceptual framework of all possible EV-virus associations and their resulting functions in infection output. First, we describe the different viral materials potentially associated with EVs by reporting that EVs can harbor entire virions, viral proteins and viral nucleic acids. We also delineate the different mechanisms underlying the internalization of these viral components into EVs. Second, we describe the potential fate of EV-associated viral material cargo by detailing how EV can circulate and target a naive cell once secreted. Finally, we itemize the different pro-viral strategies resulting from EV associations as the Trojan horse strategy, an alternative mode of viral transmission, an expansion of viral cellular tropism, a pre-emptive alteration of host cell physiology and an immunity decoy. With this conceptual overview, we aim to stimulate research on EV-virus interactions.

Mosquito transmission of dengue viruses to humans starts with infection of skin resident cells at the biting site. There is great interest in identifying transmission-enhancing factors in mosquito saliva in order to counteract them. Here we report the discovery of high levels of the anti-immune subgenomic flaviviral RNA (sfRNA) in dengue virus 2-infected mosquito saliva. We established that sfRNA is present in saliva using three different methods: northern blot, RT-qPCR and RNA sequencing. We next show that salivary sfRNA is protected in detergent-sensitive compartments, likely extracellular vesicles. In support of this hypothesis, we visualized viral RNAs in vesicles in mosquito saliva and noted a marked enrichment of signal from 3'UTR sequences, which is consistent with the presence of sfRNA. Furthermore, we show that incubation with mosquito saliva containing higher sfRNA levels results in higher virus infectivity in a human hepatoma cell line and human primary dermal fibroblasts. Transfection of 3'UTR RNA prior to DENV2 infection inhibited type I and III interferon induction and signaling, and enhanced viral replication. Therefore, we posit that sfRNA present in salivary extracellular vesicles is delivered to cells at the biting site to inhibit innate immunity and enhance dengue virus transmission.

A unique cytochrome P450 (CYP) oxidoreductase (CPR) sustains activities of human microsomal CYPs. Its function requires toggling between a closed conformation enabling electron transfers from NADPH to FAD and then FMN cofactors and open conformations forming complexes and transferring electrons to CYPs. We previously demonstrated that distinct features of the hinge region linking the FAD and FMN domain (FD) modulate conformer poses and their interactions with CYPs. Specific FD residues contribute in a CYP isoform-dependent manner to the recognition and electron transfer mechanisms that are additionally modulated by the structure of CYP-bound substrate. To obtain insights into the underlying mechanisms, we analyzed how hinge region and FD mutations influence CYP1A2-mediated caffeine metabolism. Activities, metabolite profiles, regiospecificity and coupling efficiencies were evaluated in regard to the structural features and molecular dynamics of complexes bearing alternate substrate poses at the CYP active site. Studies reveal that FD variants not only modulate CYP activities but surprisingly the regiospecificity of reactions. Computational approaches evidenced that the considered mutations are generally in close contact with residues at the FD–CYP interface, exhibiting induced fits during complexation and modified dynamics depending on caffeine presence and orientation. It was concluded that dynamic coupling between FD mutations, the complex interface and CYP active site exist consistently with the observed regiospecific alterations.

Purpose: An increasing number of anomia treatment studies have coupled traditional word retrieval accuracy outcome measures with more fine-grained analysis of word retrieval errors to allow for more comprehensive measurement of treatment-induced changes in word retrieval. The aim of this study was to examine changes in picture naming errors after phonomotor treatment. Method: Twenty-eight individuals with aphasia received 60 hr of phonomotor treatment, an intensive, phoneme-based therapy for anomia. Confrontation naming was assessed pretreatment, immediately posttreatment, and 3 months posttreatment for trained and untrained nouns. Responses were scored for accuracy and coded for error type, and error proportions of each error type (e.g., semantic, phonological, omission) were compared: pre- versus posttreatment and pretreatment versus 3 months posttreatment. Results: The group of treatment participants improved in whole-word naming accuracy on trained items and maintained their improvement. Treatment effects also generalized to untrained nouns at the maintenance testing phase. Additionally, participants demonstrated a decrease in proportions of omission and description errors on trained items immediately posttreatment. Conclusions: Along with generalized improved whole-word naming accuracy, results of the error analysis suggest that a global (i.e., both lexical-semantic and phonological) change in lexical knowledge underlies the observed changes in confrontation naming accuracy following phonomotor treatment.

Purpose: Living with the communication impairment of aphasia can be stressful. Chronic stress, depression, and anxiety are intimately linked, may be more pervasive in people with poststroke aphasia than the general population, and may influence cognitive function and treatment outcomes. In this project, we explored the psychological constructs of depression and anxiety and their associations with a biomarker measure of chronic stress in people with aphasia. Method: Fifty-seven participants with aphasia completed measures of depression and anxiety and provided a hair sample from which to extract the stress hormone cortisol. Pearson product--moment correlational analyses were used to identify associations between depression, anxiety, and long-term level of cortisol via hair sample. Conclusions: Chronic stress, depression, and anxiety have been little explored in people with aphasia to date, yet they are associated with future health consequences and impaired cognitive function, motivating further research as well as consideration of these factors in aphasia rehabilitation. Results: While cortisol level was not associated with depression and anxiety across this sample of people with aphasia, a post hoc analysis showed a significant, positive correlation between a subset of participants with moderate and higher levels of depression and elevated cortisol level.

Abstract Arboviruses such as dengue (DENV), Zika (ZIKV) and chikungunya (CHIKV) viruses infect close to half a billion people per year, and are primarily transmitted through Aedes aegypti bites. Infection-induced changes in mosquito salivary glands (SG) influence transmission by inducing antiviral immunity, which restricts virus replication in the vector, and by altering saliva composition, which influences skin infection. Here, we profiled SG proteome responses to DENV serotype 2 (DENV2), ZIKV and CHIKV infections by using high-resolution isobaric-tagged quantitative proteomics. We identified 218 proteins with putative functions in immunity, blood-feeding or related to the cellular machinery. We observed that 58, 27 and 29 proteins were regulated by DENV2, ZIKV and CHIKV infections, respectively. While the regulation patterns were mostly virus-specific, we separately depleted four uncharacterized proteins that were upregulated by all three viral infections to determine their effects on these viral infections. Our study suggests that gamma-interferon responsive lysosomal thiol-like (GILT-like) has an anti-ZIKV effect, adenosine deaminase (ADA) has an anti-CHIKV effect, salivary gland surface protein 1 (SGS1) has a pro-ZIKV effect and salivary gland broad-spectrum antiviral protein (SGBAP) has an antiviral effect against all three viruses. The comprehensive description of SG responses to three global pathogenic viruses and the identification of new restriction factors improves our understanding of the molecular mechanisms influencing transmission.

Dengue viruses (DENV) are expanding global pathogens that are transmitted through the bite of mosquitoes, mostly Aedes aegypti. As RNA viruses, DENV rely on RNA-binding proteins (RBPs) to complete their life cycle. Alternatively, RBPs can act as restriction factors that prevent DENV multiplication. While the importance of RBPs is well-supported in humans, there is a dearth of information about their influence on DENV transmission by mosquitoes. Such knowledge could be harnessed to design novel, effective interventions against DENV. Here, we successfully adapted RNA-affinity chromatography coupled with mass spectrometry-a technique initially developed in mammalian cells-to identify RBPs in Ae. aegypti cells. We identified fourteen RBPs interacting with DENV serotype 2 3'UTR, which is involved in the viral multiplication and produces subgenomic flaviviral RNA (sfRNA). We validated the RNA affinity results for two RBPs by confirming that AePur binds the 3'UTR, whereas AeStaufen interacts with both 3'UTR and sfRNA. Using in vivo functional evaluation, we determined that RBPs like AeRan, AeExoRNase, and AeRNase have pro-viral functions, whereas AeGTPase, AeAtu, and AePur have anti-viral functions in mosquitoes. Furthermore, we showed that human and mosquito Pur homologs have a shared affinity to DENV2 RNA, although the anti-viral effect is specific to the mosquito protein. Importantly, we revealed that AeStaufen mediates a reduction of gRNA and sfRNA copies in several mosquito tissues, including the salivary glands and that AeStaufen-mediated sfRNA reduction diminishes the concentration of transmission-enhancing sfRNA in saliva, thereby revealing AeStaufen's role in DENV transmission. By characterizing the first RBPs that associate with DENV2 3'UTR in mosquitoes, our study unravels new pro- and anti-viral targets for the design of novel therapeutic interventions as well as provides foundation for studying the role of RBPs in virus-vector interactions.

Mosquito saliva proteins modulate the human immune and hemostatic systems and control mosquito-borne pathogenic infections. One mechanism through which mosquito proteins may influence host immunity and hemostasis is their interactions with key human receptor proteins that may act as receptors for or coordinate attacks against invading pathogens. Here, using pull-down assays and proteomics-based mass spectrometry, we identified 11 Ae. aegypti salivary gland proteins (SGPs) (e.g., apyrase, Ae. aegypti venom allergen-1 [AaVA-1], neutrophil stimulating protein 1 [NeSt1], and D7 proteins), that interact with one or more of five human receptor proteins (cluster of differentiation 4 [CD4], CD14, CD86, dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin [DC-SIGN], and Toll-like receptor 4 [TLR4]). We focused on CD4- and DC-SIGN-interacting proteins and confirmed that CD4 directly interacts with AaVA-1, D7, and NeST1 recombinant proteins and that AaVA-1 showed a moderate interaction with DC-SIGN using ELISA. Bacteria responsive protein 1 (AgBR1), an Ae. aegypti saliva protein reported to enhance ZIKV infection in humans but that was not identified in our pull-down assay moderately interacts with CD4 in the ELISA assay. Functionally, we showed that AaVA-1 and NeST1 proteins promoted activation of CD4+ T cells. We propose the possible impact of these interactions and effects on mosquito-borne viral infections such as dengue, Zika, and chikungunya viruses. Overall, this study provides key insight into the vector-host (protein-protein) interaction network and suggests roles for these interactions in mosquito-borne viral infections.

Among emerging zoonotic pathogens, mosquito-borne viruses (MBVs) circulate between vertebrate animals and mosquitoes and represent a serious threat to humans via spillover from enzootic cycles to the human community. Active surveillance of MBVs in their vectors is therefore essential to better understand and prevent spillover and emergence, especially at the human–animal interface. In this study, we assessed the presence of MBVs using molecular and phylogenetic methods in mosquitoes collected along an ecological gradient ranging from rural urbanized areas to highland forest areas in northern Thailand. We have detected the presence of insect specific flaviviruses in our samples, and the presence of the emerging zoonotic Tembusu virus (TMUV). Reported for the first time in 1955 in Malaysia, TMUV remained for a long time in the shadow of other flaviviruses such as dengue virus or the Japanese encephalitis virus. In this study, we identified two new TMUV strains belonging to cluster 3, which seems to be endemic in rural areas of Thailand and highlighted the genetic specificities of this Thai cluster. Our results show the active circulation of this emerging flavivirus in Thailand and the need for continuous investigation on this poorly known but threatening virus in Asia.

The purpose of this study was to explore the impact of communication disorders on family members of people with Parkinson's disease (PD) through the lens of third-party disability. Nine community-dwelling family members who live with people with PD participated in semi-structured interviews. Data were collected and analyzed using the qualitative research methods from the tradition of phenomenology. Two themes emerged from the analysis of data. The first theme, "What?" captures examples of how communication between participants and their family members has changed because of PD, and how participants attempt to manage frequent communication breakdowns. Two subthemes address communication challenges related to changes in speech and changes in cognitive-linguistic skills, respectively, in their family members. The second theme, "It brings up things that have changed," addresses the broader life impact that the communication changes have had on participants. The first subtheme describes how participants are changing their roles to take on more communication-related responsibilities in their households while still trying to maintain the dignity of their family members. The second subtheme reveals how participants experience grief and loss in that the decline in communication between themselves and their family members brings about a loss of connection with the person they may be closest to and, hence, changes their relationship with that person. Family members experience third-party disability related to communication disorders in their family members with PD. Some of these experiences are similar to third-party disability experienced by family members of people with other types of communication disorders. Better understanding of the extent and nature of third-party disability would help clinicians address the communication needs of both their clients with PD and their family members.

Purpose: Chronic stress is likely a common experience among people with the language impairment of aphasia. Importantly, chronic stress reportedly alters the neural networks central to learning and memory--essential ingredients of aphasia rehabilitation. Before we can explore the influence of chronic stress on rehabilitation outcomes, we must be able to measure chronic stress in this population. The purpose of this study was to (a) modify a widely used measure of chronic stress (Perceived Stress Scale [PSS]; Cohen & Janicki-Deverts, 2012) to fit the communication needs of people with aphasia (PWA) and (b) validate the modified PSS (mPSS) with PWA. Method: Following systematic modification of the PSS (with permission), 72 PWA completed the validation portion of the study. Each participant completed the mPSS, measures of depression, anxiety, and resilience, and provided a sample of the stress hormone cortisol extracted from the hair. Pearson's product-moment correlations were used to examine associations between mPSS scores and these measures. Approximately 30% of participants completed the mPSS 1 week later to establish test-retest reliability, analyzed using an interclass correlation coefficient. Results: Significant positive correlations were evident between the reports of chronic stress and depression and anxiety. In addition, a significant inverse correlation was found between reports of chronic stress and resilience. The mPSS also showed evidence of test-retest reliability. No association was found between mPSS score and cortisol level. Conclusion: Although questions remain about the biological correlates of chronic stress in people with poststroke aphasia, significant associations between chronic stress and several psychosocial variables provide evidence of validity of this emerging measure of chronic stress.

ABSTRACT Infection with dengue virus (DENV) induces vast rearrangements of the endoplasmic reticulum, which allows the compartmentalization of viral RNA replication and particle assembly. Both processes occur in concert with viral and cellular proteins. Prior studies from our group suggest that the host RNA-binding protein (RBP) Y-box binding protein 1 (YBX1) is required for a late step in the DENV replication cycle. Here we report that YBX1 interacts with the viral nucleocapsid, distributes to DENV assembly sites and is required for efficient assembly of intracellular infectious virions and their secretion. Genetic ablation of YBX1 decreased the spatial proximity between capsid and envelope, increased the susceptibility of envelope to proteinase K mediated degradation, resulted in the formation of rough empty-looking particles, and decreased the secretion of viral particles. We propose a model wherein YBX1 enables the interaction between the viral nucleocapsid with the structural protein E, which is required for proper assembly of intracellular virus particles and their secretion. IMPORTANCE The global incidence of dengue virus (DENV) infections has steadily increased over the past decades representing an enormous challenge for public health. During infection, DENV viral RNA interacts with numerous host RNA binding proteins (RBPs) that aid viral replication and thus constitute potential molecular targets to curb infection. We recently reported that Y-box-binding protein 1 (YBX1) interacts with DENV RNA and is required at a late step of the replication cycle. Here we describe the molecular mechanism by which YBX1 mediates DENV infection. We show that YBX1 interacts with the viral nucleocapsid, distributes to DENV assembly sites and is required for efficient assembly of intracellular infectious virions. These results provide important insights into DENV assembly, revealing novel functions of host RBPs during viral infection and opening new avenues for antiviral intervention.

Summary: Dengue symptoms include alteration of blood coagulation and fibrinolysis, causing severe hemorrhage and death. Here, we demonstrate that higher concentration of plasmin, the human fibrinolytic factor, in blood meal enhances dengue virus (DENV) infection in mosquito midgut and dissemination in mosquitoes. We also show that mosquitoes express a plasmin-selective Kazal-type inhibitor (AaTI) in the midgut to inhibit plasmin proteolysis and revert the enhanced infection. Using bio-layer interferometry, we show that DENV, plasmin, and AaTI interact to form a tripartite complex. Eventually, plasmin increases midgut internalization of dextran molecules and this is reverted by AaTI. Our study demonstrates that (1) DENV recruits plasmin to increase local proteolytic activity in the midgut, thus degrading the glycocalyx and enhancing DENV internalization and (2) AaTI can act as a transmission-blocking agent by inhibiting plasmin proteolysis. Our results indicate that dengue pathogenesis enhances DENV fitness by increasing its infectivity to mosquitoes. : Virology; Entomology Subject Areas: Virology, Entomology

Mosquito-borne flaviviruses, such as dengue (DENV), Zika (ZIKV), yellow fever (YFV), West Nile (WNV), and Japanese encephalitis (JEV) viruses, threaten a large part of the human populations. In absence of therapeutics and effective vaccines against each flaviviruses, targeting viral metabolic requirements in mosquitoes may hold the key to new intervention strategies. Development of metabolomics in the last decade opened a new field of research: mosquito metabolomics. It is now clear that flaviviruses rely on mosquito lipids, especially phospholipids, for their cellular cycle and propagation. Here, we review the biosyntheses of, biochemical properties of and flaviviral interactions with mosquito phospholipids. Phospholipids are structural lipids with a polar headgroup and apolar acyl chains, enabling the formation of lipid bilayer that form plasma- and endomembranes. Phospholipids are mostly synthesized through the de novo pathway and remodeling cycle. Variations in headgroup and acyl chains influence phospholipid physicochemical properties and consequently the membrane behavior. Flaviviruses interact with cellular membranes at every step of their cellular cycle. Recent evidence demonstrates that flaviviruses reconfigure the phospholipidome in mosquitoes by regulating phospholipid syntheses to increase virus multiplication. Identifying the phospholipids involved and understanding how flaviviruses regulate these in mosquitoes is required to design new interventions.

A lipid bilayer produced from the host membrane makes up around 20% of the weight of the dengue virus (DENV) virion and is crucial for virus entry. Despite its significance, the virion’s lipid composition is still poorly understood. In tandem with lipid profiles of the cells utilised to generate the virions, this work determined a partial lipid profile of DENV virions derived from two cell lines (C6/36 and LLC-MK2). The results showed distinctive profiles between the two cell types. In the mammalian LLC-MK2 cells, 30.8% (73/237 identified lipid species; 31 upregulated, 42 downregulated) of lipid species were altered in response to infection, whilst in insect C6/36 cells only 12.0% (25/208; 19 upregulated, 6 downregulated) of lipid species showed alterations in response to infection. For virions from LLC-MK2 cells, 14 lipids were detected specifically in virions with a further seven lipids being enriched (over mock controls). For virions from C6/36 cells, 43 lipids were detected that were not seen in mock preparations, with a further 16 being specifically enriched (over mock control). These results provide the first lipid description of DENV virions produced in mammalian and mosquito cells, as well as the lipid changes in the corresponding infected cells.

Purpose: Living with the communication impairment of aphasia can be stressful. Chronic stress, depression, and anxiety are intimately linked, may be more pervasive in people with poststroke aphasia than the general population, and may influence cognitive function and treatment outcomes. In this project, we explored the psychological constructs of depression and anxiety and their associations with a biomarker measure of chronic stress in people with aphasia. Method: Fifty-seven participants with aphasia completed measures of depression and anxiety and provided a hair sample from which to extract the stress hormone cortisol. Pearson product-moment correlational analyses were used to identify associations between depression, anxiety, and long-term level of cortisol via hair sample. Results: While cortisol level was not associated with depression and anxiety across this sample of people with aphasia, a post hoc analysis showed a significant, positive correlation between a subset of participants with moderate and higher levels of depression and elevated cortisol level. Conclusions: Chronic stress, depression, and anxiety have been little explored in people with aphasia to date, yet they are associated with future health consequences and impaired cognitive function, motivating further research as well as consideration of these factors in aphasia rehabilitation., Approximately 2 million people in the United States live with aphasia, a multimodal impairment of language that can occur after stroke and other brain injuries (National Aphasia Association, n.d.). Aphasia [...]

Purpose: An increasing number of anomia treatment studies have coupled traditional word retrieval accuracy outcome measures with more fine-grained analysis of word retrieval errors to allow for more comprehensive measurement of treatment-induced changes in word retrieval. The aim of this study was to examine changes in picture naming errors after phonomotor treatment. Method: Twenty-eight individuals with aphasia received 60 hr of phonomotor treatment, an intensive, phoneme-based therapy for anomia. Confrontation naming was assessed pretreatment, immediately posttreatment, and 3 months posttreatment for trained and untrained nouns. Responses were scored for accuracy and coded for error type, and error proportions of each error type (e.g., semantic, phonological, omission) were compared: pre- versus posttreatment and pretreatment versus 3 months posttreatment. Results: The group of treatment participants improved in whole-word naming accuracy on trained items and maintained their improvement. Treatment effects also generalized to untrained nouns at the maintenance testing phase. Additionally, participants demonstrated a decrease in proportions of omission and description errors on trained items immediately posttreatment. Conclusions: Along with generalized improved whole-word naming accuracy, results of the error analysis suggest that a global (i.e., both lexical-semantic and phonological) change in lexical knowledge underlies the observed changes in confrontation naming accuracy following phonomotor treatment., The treatment of anomia, the most ubiquitous aphasia symptom, is central to aphasia therapy. Typically, the effectiveness of aphasia treatments that target anomia is examined by measuring naming accuracy of [...]