Despite possessing valuable skills, social communication differences mean that autistic people are frequently disadvantaged in job interviews. We examined how autistic and non-autistic adults compared on standard (unmodified) job interview questions, and then used these findings to develop and evaluate supportive adaptations to questions. Fifty adults  took part in two mock job interviews. Interview 1 provided a baseline measure of performance when answering typical, unmodified interview questions. Employment experts (unaware of participants’ autism diagnoses) rated all interviewees on question-specific performance and overall impressions and then provided feedback about how interviewees could improve and how questions could be adapted to facilitate this. Interviewees also provided feedback about the interview process from their perspective. Adaptations to the questions were developed, with Interview 2 taking place approximately 6 months later. Results demonstrated that, in Interview 1, employment experts rated autistic interviewees less favourably than nonautistic interviewees. Ratings of both autistic and non-autistic participants’ answers improved in Interview 2, but particularly for autistic interviewees (such that differences between autistic and non-autistic interviewees’ performance reduced in Interview 2). Employers should be aware that adaptations to job interview questions are critical to level the playing field for autistic candidates.Despite possessing valuable skills, differences in the way that autistic people understand and respond to others in social situations mean that they are frequently disadvantaged in job interviews. We examined how autistic and non-autistic adults compared on standard (unmodified) job interview questions, and then used these findings to develop and evaluate supportive adaptations to questions. Fifty adults  took part in two mock job interviews. Interview 1 provided a baseline measure of performance when answering typical, unmodified interview questions. Employment experts (unaware of participants’ autism diagnoses) rated all interviewees on their responses to each question and their overall impressions of them and then provided feedback about how interviewees could improve and how questions could be adapted to facilitate this. Interviewees also provided feedback about the interview process, from their perspective. Adaptations to the questions were developed, with Interview 2 taking place approximately 6 months later. Results demonstrated that, in Interview 1, employment experts rated autistic interviewees less favourably than non-autistic interviewees. Ratings of both autistic and non-autistic participants’ answers improved in Interview 2, but particularly for autistic interviewees (such that differences between autistic and non-autistic interviewees’ performance reduced in Interview 2). Employers should be aware that adaptations to job interview questions are critical to level the playing field for autistic candidates. The focApplicants may employ a range of IM tactics in job interviews to influence interviewer decisions see . Theory This is pertinent to autism because traditional theories posit that autistic people experience difficulties in reading others’ intentions and interpreting social cues see , 2000, wTypically, employment interviews rely upon open-ended, indirect questions such as ‘Tell me a bit about yourself’ e.g. . HoweverCritically, it has been theorised that the difficulties experienced by autistic people are most marked on open-ended test situations in which the questions or instructions do not provide the individual with an explicit understanding of the task and what is required of them . These dWhile a recently accumulating body of research has examined how autistic adults can be supported to gain employment, this work has focussed on how the interviewee can be coached to change their behaviours during interviews, neglecting changes that the interviewer can make to questions e.g. . While sIn sum, successful performance in a job interview requires effective two-way communication between an interviewer and interviewee, in order for questions to be understood in a way that enables the interviewee to formulate an appropriate response that presents themselves favourably. Unless adaptations are made that promote greater shared understanding of what is intrinsically required of an interviewee, autistic candidates are likely to be significantly limited in their ability to emphasise their best attributes and most relevant experience and overshadowed by candidates with a greater predisposition to employ IM tactics. However, employers can make positive adaptations to the interview process, particularly to the questions, which could ameliorate the disadvantage currently experienced by autistic candidates.The aim of the present study was twofold. First, to gather baseline data regarding how autistic people perform in response to standard (unadapted) employment interview questions compared with non-autistic interviewees (Phase 1). Although interviews have been identified as a major barrier to employment by autistic individuals e.g.  and theoWe predicted that differences in social communication, IM, executive functioning, and memory would result in autistic interviewees being perceived less favourably by employer raters than non-autistic interviewees in response to standard, open-ended, interview questions (in Phase 1). In line with the Task Support Hypothesis, we predicted that when more structured, explicit, and supportive questions were used (in Phase 2), their responses would be improved to the extent that the difference between groups would be ameliorated.Diagnostic and Statistical Manual of Mental Disorders  < 0.01, p = 1.00, d < 0.01, or on measures from the Wechsler Abbreviated Scale of Intelligence–Second Edition  = 0.47, p = 0.644, d = 0.13; Perceptual Reasoning Index (PRI), t(48) = 1.07, p = 0.289, d = 0.30; or Full Scale Intelligence Quotient (FSIQ), t(42.87) = 0.10, p = 0.325, d = 0.28. All non-autistic participants scored below the recommended minimum cut-off of 26 on the Autism Spectrum Quotient (AQ-50), which measures levels of autistic traits (t(40.10) = 10.29, p < 0.001, d = 2.95  years. To provide an index of their knowledge, experience, and perceptions of autism, employers also completed brief scales. These demonstrated scores within the average range to those previously reported: Autism Awareness Scale  were recruited via the researchers’ professional contacts (within the employment industry) to rate transcripts of participants’ answers. The raters worked in various roles for different companies  and all had substantial experience in recruiting and interviewing. Their ages ranged from 38 to 52  × 2 (Phase: Phase 1 unadapted questions vs Phase 2 adapted questions) mixed design, where Phase was within participants. In Phase 1, all participants answered standard employment interview questions from one of two interview schedules (A or B). In Phase 2, the same participants returned to answer adapted interview questions from the interview schedule they did not receive at Phase 1. Dependent variables were employment professionals’ quantitative scale ratings of interviewees’ answers to each of the seven interview questions and their overall impression of the interviewee measured on nine aspects of participants’ overall performance (see below for details). Qualitative feedback was also obtained from both employers and interviewees to inform the development of question adaptations for Phase 2 and provide a more in-depth exploration of the efficacy of the different question types.Supplemental Appendix A).Two interview schedules (A and B) were developed, each with seven questions typical of standard employment interviews. Questions included those aimed at eliciting descriptions of experience and activity, personality characteristics, and self-evaluative information (e.g. ‘What are some of your strengths?’), as well as past job experience and situational judgements  to 5 (excellent) to provide a more fine-grained distinction between those who barely answered the question at all  and those who did respond, but poorly . To inform the development of adaptations to questions for Phase 2, employers were also asked two optional open-ended questions relating to each interviewee’s response to each interview question in Phase 1: ‘How could the interviewee improve their answer?’ and ‘How could the question be adapted to support this?’ .Employment professionals’ ratings of the quality of interviewees’ responses to each question were scored using an adapted form of the Interview Skills Rating Instrument . The orinot at all to extremely. These constructs were identified in a review by 2 , as well as which questions they found easy/challenging and why, and what would have made the interview easier for them. In addition, at Phase 2, interviewees were also asked whether they used the printout of the questions (details below) and how useful they found this, as well as how this second interview compared with the interview that they had received in Phase 1.At the end of each interview, interviewees completed an online Qualtrics survey about their experience of the interview. This included questions about their confidence in their performance and how clear the desired/required responses were from the questions . Interviewees were asked each of the seven interview questions in the same order. In cases where participants could not provide an answer, they were given plenty of time, but the interviewer moved on to the next question if they were still unable to answer (they could not return to the unanswered question).3 The interviewer provided minimal verbal feedback throughout, but the question was repeated or clarified if requested (limited to rephrasing the question or using synonyms). After the interview, participants completed the online feedback survey asking them about their experience of the interview.Participants completed the mock employment interview in a quiet room at the University of Bath. All participants were interviewed by the second author (J.E.N.) in the same room. Participants were not provided with a specific job description (for parity across participants with different levels of skills and experience), but were asked to answer the questions as though they were in a real employment interview. Participants were pseudo-randomly allocated to receive either schedule A or B at Phase 1, within the constraints of ensuring groups were matched on age and intelligence quotient (IQ)  to receive adapted questions from the alternative interview schedule to that completed at Phase 1. Again, they were told that there was no specific job description and that they should answer the questions as though in a real employment interview. The interviews were conducted in the same way as at Phase 1 , with three exceptions. First, to negate any potential increases in executive demands due to the multi-part nature of the adapted questions, participants were asked to respond to each part of the question in turn. Second, and relatedly, they were provided with a printout of the questions, which remained in front of them throughout the interview. Third, interviewees were given more explicit instructions regarding the structure of the interview, such as the number of questions they were going to be asked and when they should provide an answer. At the end of the interview, participants repeated the online feedback survey about their experience of the interview.Participants returned around 6 months later .www.asmentoring.co.uk).Results are reported by phase. For Phase 1 (when standard questions were used), between-group comparisons examined whether employers’ quantitative ratings differed between autistic and non-autistic interviewees in terms of (1) question-specific performance and (2) their overall impressions of them. Phase 2 is presented in two parts. First, a development stage is reported in which a content analysis of employers’ qualitative feedback from Phase 1 about how interviewees’ performance could be improved and how questions could be adapted to support this (conducted by J.E.N. and J.N.) and interviewees’ reflections on their Phase 1 interview experiences (conducted by B.H.) was used to guide the development of adaptations to questions for Phase 2. This involved an initial inductive content analysis , wherebyPhase 2 interview data  were analysed using Group × Phase mixed models. These tested the effects of the adapted questions on employers’ ratings of autistic versus non-autistic interviewees’ performance and their overall impressions of them. A content analysis (conducted by B.H.) then examined how the adapted interview questions affected the interview experience as reported by autistic and non-autistic interviewees, while mixed analyses of variance (ANOVAs) tested whether the adapted questions improved autistic and non-autistic interviewees’ ratings of confidence in their performance and how clear they felt the desired responses were from the questions.SDs below the mean). This participant was removed from the subsequent quantitative analyses of employers’ ratings (NB. the pattern of findings remained the same). Interview schedule (A or B) had no effect on employer ratings of interviewees’ responses to questions or on their overall impression ratings in either Phase 1 or Phase 2 . Subsequent analyses were therefore collapsed across interview schedule.Inspection of employer ratings data revealed an outlier in the autistic group who received unusually low rating scores across all measures  than their ratings of non-autistic interviewees’ responses , t(47) = 3.61, p = 0.001, d = 1.03  = 4.32, p = 0.001, Fs > 7.58, ps < 0.008, Fs < 3.64, ps > 0.063, A multivariate analysis of variance (MANOVA), with Group as the independent variable and mean ratings on each of the nine impressions items as the dependent variables, indicated a significant multivariate effect of Group on employers’ overall impressions of interviewees in Phase 1, Pillai’s Trace = 0.50, .14) see . Groups  < 0.07) .Content: General, (2) Content: Self-reflection, (3) Focus, (4) Length, and (5) Delivery . They also suggested that interviewees should show more positive self-reflection in their responses by reducing negative comments made about themselves and highlighting what they learnt from situations. Employers also recommended that autistic (and to a lesser extent non-autistic) interviewees needed to focus, think before responding, maintain clarity, and directly answer the question. Many answers provided by both autistic and non-autistic interviewees were criticised for lacking structure and for inappropriate length (too short/too long). Employers also suggested that both autistic and non-autistic interviewees’ delivery needed to be more positive/confident and that autistic interviewees’ responses in particular would be improved if they were less hesitant.The major theme categories that were identified from the content analysis of employers’ suggestions for how interviewees could improve their answers fell under (1) very see . In summprompt for specific information and self-reflection (e.g. what the interviewee learned from a situation). Employers also frequently suggested that interviewees should be given more guidance on how to respond and how the question structure could be adapted to facilitate this (e.g. by asking questions in multiple parts).Main themes for employers’ suggested adaptations were identified for each question from schedules A and B . Codes broadly fell under one of four over-arching themes . Overallself-reflections, (2) interpersonal dynamic, (3) question clarity and structure, and (4) adaptations. self-reflected on aspects of their performance relating to cognition, emotion, and communication. The autistic group reported more cognitive difficulties  and slightly higher communication challenges  compared with the non-autistic group. Within the interpersonal dynamic of the interview, both groups reported using strategies to manage impressions, but the non-autistic group were more likely to report concerns about how to ‘sell’ themselves, downplaying their difficulties and boosting their strengths, while the autistic group expressed concerns about being too honest.The content analysis resulted in four organising themes: (1) question clarity and structure, autistic interviewees were more likely to report that questions were unclear and poorly structured than the non-autistic group. Accordingly, both autistic and non-autistic interviewees suggested that adaptations should include the use of more specific questions. Autistic interviewees felt that visual supports  would be particularly helpful. Both groups also reported a wish for questions with more flexible answers (e.g. that enabled them to draw more widely on their experiences), more feedback on questions, and having more time to prepare and respond to questions.Reflecting on Supplemental Appendix A for the full list of adapted questions). All interviewees were also provided with printouts of the adapted questions in Phase 2, which were visible throughout the interview to further support their comprehension of questions, in line with previous suggestions by both employers and interviewees.5Questions were adapted based on the qualitative analyses of employer and interviewee feedback, primarily aiming to make the desired response more explicit, for example, by requesting more specific information and details and supporting the interviewee to structure their answer clearly with effective use of examples. This was achieved by adapting the question wording such that the interviewee was first oriented to the topic (to set the context), before breaking the question down into separate, specific questions (to request particular details while avoiding compound questions with multiple and/or stacked clauses). F = 7.37, p = 0.010, M = 3.62, SD = 0.44) than non-autistic interviewees’ responses overall . There was also a main effect of Phase, F = 39.65, p < 0.001, M = 3.97, SD = 0.44) than at Phase 1 . Crucially, there was a Group × Phase interaction, F = 5.03, p = 0.031, p = 0.010, d = 0.85), the difference between groups at Phase 2 was not significant .A 2 (Group) × 2 (Phase) mixed ANOVA was used to examine whether the question adaptations improved the quality of autistic and non-autistic interviewees’ responses (the dependent variable was employers’ mean ratings of interviewees’ responses to the seven questions within each phase). There was a significant main effect of Group, F = 4.13, p = 0.001, F = 3.63, p = 0.003, F = 1.25, p = 0.303, A 2 (Group) × 2 (Phase) mixed MANOVA examined whether employers’ overall impressions of interviewees on each of the nine items improved following the adaptations to questions. There were significant multivariate effects of Group, Pillai’s Trace = 0.55, Fs > 4.54, ps < 0.039, Fs < 2.87, ps > 0.098, Separate univariate tests on the outcome variables revealed significant effects of Group (with autistic interviewees again being rated less favourably) in terms of confidence, communication skills, likeability, and perceived ease to work with  .Fs > 4.21, ps < 0.047, Fs < 1.69, ps > 0.202, Univariate tests on the outcome variables revealed that employers’ overall impressions of interviewees’ conscientiousness, competence, communication skills, likeability, and ease to work with all improved with the provision of adaptations to the questions in Phase 2  .Interviewees’ reflections on their interview experience in Phase 2 were coded by B.H. using the content analysis process as outlined for Phase 1. F = 4.12, p = 0.049, M = 3.55, SD = 0.77) compared with Phase 1 . There was no main effect of Group, F = 3.23, p = 0.080, F = 1.98, p = 0.168, F = 17.68, p < 0.001, M = 4.40, SD = 0.59) compared with Phase 1 . There was no main effect of Group, F = 2.79, p = 0.103, F = 5.03, p = 0.031, M = 3.31, SD = 0.74) than non-autistic participants  at Phase 1 (p = 0.009), there was no difference between groups in reported clarity of questions at Phase 2 (p = 0.843).Finally, a 2 (Group) × 2 (Phase) mixed ANOVA on interviewees’ ratings of their confidence in their performance indicated a main effect of Phase, employers can make to their questions, rather than the onus being entirely on the autistic person to adapt their interview technique. It is also, to our knowledge, the first study to compare how autistic adults perform against non-autistic adults during employment interviews; this is important given that, in most real-life job interview scenarios, autistic and non-autistic candidates will be competing with one another.The current study builds upon previous research on supporting autistic people in employment interviews e.g. . The focWhen standard (unadapted) interview questions were used in Phase 1, employment professionals provided lower ratings for both the quality of autistic interviewees’ answers and their overall impressions of them compared with non-autistic participants. Following adaptations to the questions, there was a significant improvement on both of these measures. While autistic participants were still rated less favourably than non-autistic participants on overall impressions in Phase 2, they showed a greater improvement in their answer quality than the non-autistic group, to the extent that differences between the groups were reduced in Phase 2. It is worth noting that there remained a marginally significant difference between groups with a medium effect size, which warrants further detailed examination. Employers’ ratings of their overall impressions of interviewees tentatively suggest that this disparity may relate to autistic interviewees being perceived as having poorer communication skills and appearing less confident. Overall, these results highlight the potential utility of combined interventions that take a two-pronged approach by focusing on training for both interviewees and interviewers see . They alAs suggested by The present study developed novel adaptations to questions to support autistic difficulties and differences in IM see , self-moIt may be critical that adaptations were made not only to the questions themselves but also to the way they were asked. Because autistic individuals often experience difficulties in executive function together with language issues that are more apparent when processing longer and more complex sentences e.g. , the intAdaptations also improved employers’ overall impressions of both autistic and non-autistic interviewees in terms of their conscientiousness, communication skills, likeability, and ease to work with. Nevertheless, autistic interviewees were still rated more negatively than non-autistic interviewees (across both phases) in terms of their overall confidence, communication skills, likeability, and perceived ease to work with. This is somewhat surprising given that employers rated transcripts, rather than videos, of interviewees: It may also be pertinent that employers in the present study were not informed of interviewees’ diagnoses. Previous research shows that disclosing one’s autism diagnosis can have a profound effect in improving others’ perceptions of them both in everyday contexts e.g.  and in sThe current study has several limitations that are important to acknowledge. First, participants completed a mock interview scenario with no specific job description. Although this promoted parity across participants from a wide range of backgrounds and abilities , it nevertheless limited ecological validity. In particular, since role-specific knowledge and confidence in one’s abilities to successfully perform within that particular role are important drivers of interview performance see , particiquality of interviewees’ responses (i.e. their content) without the confounds of behavioural cues such as demeanour. However, this precludes a more holistic understanding of how autistic candidates are perceived in employment interviews, which is pertinent given that non-verbal behaviours are also known factors in perceived job interview performance (e.g. Second, and relatedly, the current findings are based upon employer perceptions of interviewees based solely on interview transcripts. Focussing only on the content of their responses was deemed important in this initial exploratory investigation to test the effects of adaptations in improving the nce e.g. . The finThird, interviewer feedback was limited to ensure consistency across participants, thus restricting the social dynamics of the interview and, arguably, ecological validity. Yet, previous research has shown that highly structured employment interviews are more reliable and valid than unstructured interviews as they control different biases, making the same job-related information salient to all interviewers and helping to ensure that applicants are rated consistently across interviewers .Finally, autistic adults experience higher rates of co-occurring emotional disorders such as anxiety and depression , which aIt is worth mentioning that participants in the current study also fed back that offering preparation time by providing the questions in advance of the interview would be another helpful adaptation see also . For theWhile further inquiry is required along a number of avenues such as those outlined above, there are also more immediate implications from the current preliminary study. Most notably, that interview questions should be more explicit and specific in requesting the information that is required from an interviewee, rather than utilising standard underspecified questions that rely on candidates ‘reading between the lines’ to respond desirably with the requisite information. Breaking questions down into their component parts and providing a printout of questions can also be beneficial – for both autistic and non-autistic candidates. The onus of employment interventions to date has been predominantly targeted at autistic people, rather than on changes that employers can make to better enable them . The preIn conclusion, the current study provides the first test of interview question adaptations that employers can make for autistic candidates, rather than placing the responsibility solely on the autistic interviewee. Importantly, the adaptations were also effective at improving the interview performance of non-autistic participants, embodying the principles of universal design . FindingClick here for additional data file.Supplemental material, sj-pdf-1-aut-10.1177_1362361320981319 for Ameliorating the disadvantage for autistic job seekers: An initial evaluation of adapted employment interview questions by Katie Maras, Jade Eloise Norris, Jemma Nicholson, Brett Heasman, Anna Remington and Laura Crane in Autism


A facile method to produce few-layer graphene (FLG) nanosheets is developed using protein-assisted mechanical exfoliation. The predominant shear forces that are generated in a planetary ball mill facilitate the exfoliation of graphene layers from graphite flakes. The process employs a commonly known protein, bovine serum albumin (BSA), which not only acts as an effective exfoliation agent but also provides stability by preventing restacking of the graphene layers. The latter is demonstrated by the excellent long-term dispersibility of exfoliated graphene in an aqueous BSA solution, which exemplifies a common biological medium. The development of such potentially scalable and toxin-free methods is critical for producing cost-effective biocompatible graphene, enabling numerous possible biomedical and biological applications. A methodical study was performed to identify the effect of time and varying concentrations of BSA towards graphene exfoliation. The fabricated product has been characterized using Raman spectroscopy, powder X-ray diffraction, transmission electron microscopy and scanning electron microscopy. The BSA-FLG dispersion was then placed in media containing Astrocyte cells to check for cytotoxicity. It was found that lower concentrations of BSA-FLG dispersion had only minute cytotoxic effects on the Astrocyte cells. Graphgle atom ,3,4. The V−1 s−1 , accredi V−1 s−1 . Despite V−1 s−1 ,8. A var V−1 s−1 –11 and m [et al.  used gra [et al.  discoverMany methods have been developed to fabricate graphene since the last decade. The prominent ones can be broadly classified into—(i) epitaxial growth of graphene , (ii) miExfoliation using mechanical methods has generally helped develop high-quality graphene. Nevertheless, there is still a lot of work to be done in order to improve the efficiency of the process . The exfet al. produced graphene by sonicating graphite in organic solvents including N-N-dimethylformamide (DMF) and N-methylpyrrolidone (NMP), and the material was centrifuged after sonication. The limitation of this facile technique was that the graphene yield obtained turned out to be merely 0.01 mg ml−1 [et al. [Highly ordered pyrolytic graphite (HOPG) was used to produce graphene via micromechanical cleavage ,24,25. T mg ml−1 . Buzaglo [et al.  devised  [et al. –29 makina,b). This method yields graphene flakes of large size. However, during the milling process, it is probable that the balls may strike the graphite/graphene flakes normal to the surface causing them to reduce in lateral dimensions. Moreover, the crystal structure may also be adversely affected leading to the formation of amorphous material [The principle behind the fabrication of graphene using ball milling is that the application of shear force on graphite by the milling balls leads to its exfoliation a,b. Thismaterial . The balmaterial  and planmaterial . Stirrinmaterial .Figure et al. produced hydrophilic graphene dispersions by ball milling graphite for 4 h at 400 r.p.m. along with poly(vinylpyrrolidone) and supercritical CO2, using a stirring ball milling machine [2 greatly assists in intercalation for it possesses a molecule size of 0.33 nm, comparable to the graphene interlayer distance of 0.34 nm [2 requires relatively low pressure and temperature to reach a critical point [et al. produced graphene by ball milling graphite with dry ice (solid CO2) in a planetary mill for 24 h. The resulting powder was sonicated for 20 min in the presence of 1 M aqueous HCl solution leading to the formation of edge-carboxylated graphene (ECG). Thermally reduced graphene (TRG) was obtained by annealing the ECG at a temperature of 900°C for a time duration of 10 min [et al. exfoliated graphene using 2-ethylhexanol and kerosene as solvents. A planetary ball mill was employed to conduct the high-energy milling operation for a duration of 60 h at 400 r.p.m. Centrifugation along with heat treatment was performed in the presence of Argon at 600°C [et al. conducted ball milling to exfoliate multi-layer graphite nanosheets into graphene. The solvent used was N,N-dimethylformamide (DMF) [et al. developed few-layer graphene by milling graphite along with melamine using a planetary ball mill. The parameters for the graphene samples produced, speed of rotation and milling time, ranged between 100–250 r.p.m. and 30–60 min, respectively. The experiments conducted were either in the presence of nitrogen or a normal atmosphere [et al. produced graphene using a similar process, ball milling it in the presence of melamine at 100 r.p.m. for 30 min. The drawback was that their process took a time span of 6–7 days, for excess melamine had to be removed from graphene to meet safety constraints [Chen  machine . Supercr machine . CO2 greal point . Similarat 600°C . These tde (DMF) . In thisde (DMF) . Leon etmosphere . Althougstraints ,40.et al. milled graphite at 250 r.p.m. in the presence of carbohydrate to produce graphene dispersions. They found that glucose showed the highest efficacy in exfoliating graphene with a reduced presence of defects [et al. [et al. produced graphene using a kitchen blender after applying shear/turbulence force on graphite in the presence of BSA. They also explored the relation between varying BSA–graphite ratios on the exfoliation rate and came to the conclusion that the best results could be obtained from a ratio of 0.03 [Lately, the influence of biological materials as exfoliates has also been investigated –44. Gonz defects . Ahadian [et al.  develope [et al. . Pattamm of 0.03 .et al. [Ahadian et al.  establiset al. .This paper describes a green one-step technique to fabricate biocompatible graphene using a planetary ball mill. One of the objectives of this experimental study was to investigate techniques to facilitate the fabrication of mass-produced graphene; therefore, the aqueous dispersion of FLG produced is not centrifuged after it has been collected. Moreover, to use the beneficial effects that BSA lends to FLG's biocompatibility, these two materials are not separated in any post-processing step.. 2ρ ∼ 7.83 g cm−3), each weighing approximately 8.3 g were placed in each of the steel jars.BSA (CAS: 9048-46-8) and graphite  were purchased from Sigma Aldrich USA. The concentration of graphite was kept constant at 100 mg in each experiment throughout the study. The concentration of BSA was varied according to the design of the experiment. In addition, 5 ml of water was added to the graphite–BSA mixture to prepare a solution. A two-station horizontal planetary mill  was used for the mechanochemical processing. The milling containers were made of 316L grade stainless steel. The speed of rotation was kept constant at 300 r.p.m., with the milling jars being kept at room temperature. The direction of rotation alternated along with intermittent pauses to prevent overheating. Sixteen chrome steel balls  had smooth textures. Whereas FLG that was exfoliated in the presence of large amounts of BSA  displayed a foamy texture and usually had to be scooped out using a spatula . Additionally, the dispersions with large concentrations of BSA were found to disperse for several days. 3.1n-n stacked layers and is indicated by the decreasing of the (002) peak intensity. Furthermore, this demonstrates that the size of the graphite particles is reducing normal to the basal plane. Also, there is an increase in the 002 peak's full width at half maxima (FWHM) due to Scherrer broadening along with the reduction in peak intensity [X-ray diffraction was primarily used to investigate the exfoliation of graphene. The reduction in the size of graphite flakes is caused due to the diminishing of the ntensity ,49.θ peaks (a–d). This could be attributed to the fact that graphite particles are randomly oriented which results in the flakes experiencing a combination of compressive and shear forces, which might cancel each other.While ball milling time was a contributor to the exfoliation of graphene sheets, it was found that ball milling for shorter durations of time produced sharp 2θ peaks a–d. Thisθ peaks produced were broad and graphene like (a–d). Also, this is the first time that graphene has been exfoliated in the absence of any intercalating agent by ball milling. However, the application of such prolonged times can be considered uncompetitive for industrial applications. Also, samples milled for long periods of time can get contaminated, especially from materials present in the balls as well as the milling jar. It was observed that the graphene produced after milling for 90 h, without BSA, was magnetic, which was attributed to iron contamination from the steel jars.Additionally, the individual graphene layers generally restack unless there is an intercalation agent preventing it ,50. It wene like a–d. AlsoMoreover, increasing the initial concentration of BSA helped expedite the separation of graphene sheets. The effect of BSA in the exfoliation of graphene is prominent for shorter milling times, although its effects are subtly visible for longer periods. Furthermore, XRD of pure BSA and 45 h milled BSA was performed to analyse whether milling led to the formation of amorphous BSA. One characteristic XRD peak of BSA disappeared after the milling process, making it difficult to reach a conclusion of the nature of the material after milling was performed .. 3.2a,b).The initial experimental design of this study was to investigate the effects of specific ball milling times with the increase in the initial concentration of BSA. However, after noting the impactful effects of BSA in the exfoliation of graphene, the milling time was drastically cut short to further optimize the process. Milling equal concentrations of BSA and graphite for an hour gave significantly better results than those obtained after 10 h in the absence of BSA a,b.Figu. 3.3a,b). Multiple runs (spectrum 1 and 2) were performed on different samples to obtain a fair estimate of the contamination. The sample contains approximately 3.5 atomic % impurity consisting of mainly Fe and Cr from the milling vials as shown in For purity critical applications, it is imperative that the synthesized graphene be investigated for materials that are present in the milling jar after long mechanochemical operations. Therefore, energy-dispersive X-ray spectroscopy (EDS) was performed to gauge the level of contamination in the synthesized sample, milled for 45 h a,b. Mult. 3.4a,b). Further TEM images at different magnifications have been included as supporting information .Transmission electron microscopy (TEM) was employed to further discern the structure of the ball-milled material. The FLG was in the form of folded nanosheets; however, the sample containing BSA was found to contain monolayer graphene. Moreover, it can be verified that despite the same number of milling hours, BSA-FLGs have a lower number of layers than those without BSA a,b. Furt. 3.5−1 [et al. collected Raman spectra using 514.5 nm laser excitation on graphite and graphene. The G peak and the 2D band were acutely observed at approximately 1580 and 2700 cm−1, respectively [−1 and has been referred to as the 2D' peak [The effect of ball milling on the production of defects and graphene exfoliation was determined using Raman spectroscopy. The D and G peaks are the prominent peaks observed in the Raman spectra of carbon materials and other polyaromatic hydrocarbons ,52. Thes−1 ,53,54. T−1 –46. Ferrectively ,56. An a2D' peak .D/IGI, a numerical value denoting the production of defects, ranged from 0.48 to 1.19, the latter arising after grinding was performed for 90 h . González et al. obtained an D/IGI of approximately 1.8 after milling for 8 h using glucose. However, even after milling for 90 h using BSA, the BSA-FLG has lower defect ratios than that produced using glucose as an exfoliating agent [et al. discovered that the D/IGI ratios reached up to 0.6 after exfoliating graphene in the presence of BSA using the shearing force applied by a kitchen blender. It can be inferred that any application of shear force for the production of graphene tends to generate defects in the material [The defects produced due to the milling process are overall directly proportional to the number of hours the process was carried out. However, no clear trend could be discerned to describe the effect of BSA on the formation of defects. The for 90 h a,b. Gonzng agent . Additiomaterial .Figure DI/GI. The qualitative control of structural transformation in various graphitic materials can be calibrated by Tuinstra and Koenig's (T-K) empirical relation [Dn. The nanographite size  stress. However, they found that hydrophilic graphene did not exhibit the harmful traits that hydrophobic graphene possessed. They concluded that surface functionalization is one of the salient factors that mitigate the cytotoxic characteristics of pristine graphene [a–d).Astrocytes are a type of glial cell abundantly found in the central nervous system (CNS). They perform a variety of functions including axon guidance, synaptic support, control blood–brain barrier and are very responsive to CNS attacks . Sasidhagraphene –64. Sinc. 4This paper investigates a mechanochemical process to fabricate biocompatible graphene. The effect of BSA on the accelerated exfoliation of graphene while producing relatively lower defects can be easily discerned. Additionally, this paper shows the production of graphene in the absence of any exfoliating agent after milling for large time periods. Preliminary investigations found that lower concentrations of BSA-functionalized graphene can be integrated with cells without inducing their death. Future work can include investigations of the optimal amount of BSA that allow high concentrations of graphene to be placed within the cell media. This approach may be applied in areas that require a cheap one-step fabrication method for FLG while recognizing the impact of milling speed and BSA–graphite concentrations.. 5. 5.1α1 radiation was employed with a 0.02° 2θ step, in the 2θ range from 10° to 80°. In order to prevent the characterized sample from being exposed to oxygen and moisture, a polyimide (Kapton) film was implemented. This causes the generation of amorphous-like background at 13° ≤ 2θ ≤ 20°. The powders of FLG were obtained by drying the suspension in an oven kept at 60°C. The as-dried powder was fixed on the zero-background silicon substrate using vacuum grease.The powder X-ray diffraction (PXRD) analysis was performed at room temperature using a PANalytical X'PERT diffractometer. Cu-K. 5.2The SEM images were taken using FEI Teneo Lovac FE-SEM, using an accelerating voltage of 10 kV and reported beam current of 1.6 nA. The powders of FLG were obtained by drying the suspension in an oven kept at 60°C. The as-dried powder was sprinkled sparingly on conductive carbon tape.. 5.3The EDS analysis was performed using an Oxford Aztec system with X-Max 80 detector, attached to the Teneo.. 5.4The TEM images were taken using a JEOL 2100 scanning and electron microscope. A Gatan OneView 4 K camera was used to capture images. The sample preparation included pipetting 2 µl of the aqueous FLG dispersion onto a carbon film copper grid. The excess material was removed using a filter paper and a thin film of the material was used for characterization.. 5.5The pictures of cells were captured using an inverted microscope platform . The samples were prepared using BSA-FLG at varying concentrations. Propidium iodide staining was conducted to identify cell death.. 5.6−1 with a 600 grooves mm−1 grating, each spectrum corresponds to an average of three measurements with a 30 s acquisition time and two accumulations. Sample preparation involved putting a drop on a silicon wafer and letting it dry overnight.Raman spectra were collected using a Horiba XploRa Plus confocal Raman upright microscope equipped with a 532 nm excitation source (1.5 mW at the sample) and a Synapse EMCCD camera. A 50 x air objective  with 0.25 numerical aperture was used to collect Raman spectra in the epi-direction. The spectra were collected from 600–3300 cmClick here for additional data file.


MoSe2/graphene heterojunctions with low-doped W compositions were synthesized by a one-step hydrothermal catalysis approach. Based on the conducted density functional theory (DFT) calculations, it was determined that inserting a small amount of W (≈5%) into the MoSe2/graphene heterostructure resulted in the modification of its lattice structure. Additionally, an increase in the distance between layers (≈8%) and a decrease in the adsorption energy of the potassium ions (K+) (≈−1.08 eV) were observed following W doping. Overall, the electrochemical performance of the MoSe2/graphene hybrid was enhanced by the presence of W. An all-solid-state supercapacitor device prepared using electrodes based on the W-doped MoSe2/graphene composite achieved excellent capacitance of 444.4 mF cm−2 at 1 mV s−1. The results obtained herein revealed that the MoSe2/graphene hybrid exhibiting low W composition could be valuable in the field of energy storage and isoelectronic doping of TMDs.Transition metal dichalcogenides (TMDs) have attracted widespread attention due to their excellent electrochemical and catalytic properties. In this work, a tungsten (W)-modulated molybdenum selenide (MoSe Owing to the rapid decrease in fossil fuels and increasing energy demand, the development of simple and effective energy sources is essential ,2,3. Sup2) is a particularly important material displaying a large layer spacing (0.646 nm), which decreases the energy barrier for adsorption/desorption of charged ions ·H2O, and 0.2 g of Se were mixed with 5–10 mL of N2H4·H2O and stirred for 15 min.  Fifty milliliters of deionized water was added, and the solution was stirred for 15 min. After mixing, the mixture was divided into two equal parts and transferred to two 60 mL Teflon-lined stainless steel autoclaves. The prepared nickel foam/graphene was subsequently placed in one of them. The mixture was heated to 160 °C for 24 h. The sample containing the nickel foam/graphene was then taken out and washed several times with deionized water and ethanol. The sample was dried at 80 °C for 24 h under a vacuum. Ten milliliters of the reaction solution was added into another container, centrifuged, and ultrasonicated. At the same time, different Mo: W ratios  and various temperatures  were investigated to study the impact on the performance of composite electrodes.Point three grams of Na2/graphene nanostructures were investigated by SEM . The crystallinity of the materials was evaluated by high-resolution XRD  analysis in the Bragg (reflection) geometry with a pure Cu kα1 radiation . The Raman spectrum was obtained with a Raman spectroscopy system  with an Ar+ laser (514 nm wavelength) as the excitation source. The elemental analysis of the composite was conducted by XPS .The W-doped MoSeFive grams of PVA  was added to 35 mL of deionized water. The solution was stirred in a water bath at 80 °C until the PVA was completely dissolved. Subsequently, 5 g of KOH was dissolved in 5 mL of deionized water. The KOH solution was slowly added to the solution of PVA and stirred.−1. Constant current charging/discharging was performed at a current density of 0.5 to 8 mA cm−2. EIS was conducted in the frequency range of 0.01 Hz to 100 kHz with a disturbance of 5 mV (rms) under open-circuit voltage. A three-electrode test system was used for the electrochemical measurements. The system was composed of an active material as the working electrode, Ag/AgCl as the reference electrode, platinum mesh as the counter electrode, and 3 M KOH as the electrolyte. For the electrochemical measurements of the all-solid-state supercapacitors, the electrodes and cellulose membrane were stacked in a sequence. The active material was in direct contact with the current collector. The electrolyte employed for the analysis was PVA–KOH. CV was conducted at a scan rate of 1 to 50 mV s2), V indicates the scan rate (vs−1), and △V is the potential window (V).For the CV curves, the specific capacitance values were calculated using Equation (1):For the GCD curves, the specific capacitance was calculated according to Equation (2):−2), P indicates the power density (mW cm−2), and Δt (s) is the total discharge duration.The energy density and power density of the supercapacitor cell were obtained using Equations (3) and (4):2/graphene containing low W contents are illustrated in 2. When the Mo atoms were substituted by W, the doping concentration of W was 5.6%, which was consistent with the results obtained by X-ray photoelectron spectroscopy (XPS) (5%). + adsorption energy of MoSe2/graphene doped with different concentrations of W . Compared with pure MoSe2/graphene, MoSe2/graphene doped with W at a concentration of 5.6% exhibited a larger interlayer distance  and smaller K+ adsorption energy (−1.08 eV). This facilitated the process of K+ adsorption/extraction. The adsorption energy of K ions (2 and graphene is higher than that between W-doped MoSe2. This may be the interlayer distance between graphene and W-doped MoSe2(≈ 5.6 Å) is smaller than that between tungsten-doped molybdenum selenide, making it more difficult for potassium ions to insert/detach.The structures of MoSe2/graphene nanostructures  and energy dispersive spectrometry (EDS) were used to characterize the W-doped MoSeructures a–c. It wructures a, while oflowers b,c. The 2/graphene nanostructures are shown in −1 corresponded to the G and 2D bands of graphene, respectively. In addition, the signals at 244.2 and 287.5 cm−1 were attributed to the A1g and E2g1 modes, correspondingly. A slight shift of the peaks to the lower wavenumbers compared to those of pure MoSe2 confirmed the successful doping of W in MoSe2 [2 nanostructure, respectively. The detected peaks were comparable to those of pure MoSe2; however, a small shift to lower values was noted. The particle size of the powdered samples was not uniform enough, and there were some by-products in the hydrothermal synthesis, resulting in a lot of XRD data miscellaneous peaks, such as Mo3Se4, Mo15Se19, and Se rings [2/graphene/Ni foam electrode samples. The electrode was washed several times with ethanol and deionized water and dried in a vacuum oven at 60 °C for 12 h, in order to obtain a well-crystallized product. The prepared materials are shown in 2(PDF#29-0914). A slight shift in the peaks to the lower wavenumbers compared to those of pure MoSe2 confirmed doping of W in MoSe2. The peaks at 43.3°, 51.8°, and 81.4° of graphene position corresponding to the (1 0 0), (1 0 2), and (1 1 2) crystal (PDF#41-1487). Furthermore, XPS was employed to investigate the elemental composition of the W-doped MoSe2 nanostructure   h. The ractively) i could b2/graphene electrodes was evaluated by cyclic voltammetry (CV), galvanostatic charge–discharge (GCD), and electrochemical impedance spectroscopy (EIS) using 3 M KOH as the electrolyte. Different ratios of raw materials , growth time , and temperature  were tested to achieve the most optimal conditions for the growth the W-doped MoSe2/graphene heterostructure. −1. The largest area of mMo:mW = 1:1 under the CV curve of the electrode indicated the highest capacitance. −1. The electrode grown at 160 °C displayed a longer discharge time, suggesting higher capacitance. Moreover, varying the growth time of the electrode demonstrated that the best results were achieved after 24 h. The material obtained after this time exhibited the largest capacitance –KOH electrolyte and W-doped MoSeoperties f [27,28,2/graphene heterostructures containing low contents of W were successfully prepared by a catalytic hydrothermal method. DFT calculations were used to evaluate the characteristics of W-doped MoSe2/graphene, which exhibited a large interlayer distance and low K+ adsorption energy. This facilitated electrochemical reactions. Moreover, electrochemical investigations demonstrated that all-solid-state supercapacitors based on the MoSe2/graphene heterostructure displayed excellent supercapacitor performance. Specific capacity of 444.4 mF cm−2 was achieved at a scanning rate of 1 mV s−1. Notably, 81.3% of capacitance was retained after 5000 cycles at 500 mV s−1. In conclusion, MoSe


Papillomavirus L1 and L2, the major and minor capsid proteins, play significant roles in viral assembly, entry, and propagation. In the current study, we investigate the impact of L1 and L2 on viral life cycle and tumor growth with a newly established mouse papillomavirus (MmuPV1) infection model. MmuPV1 L1 knockout, L2 knockout, and L1 plus L2 knockout mutant genomes  were generated. The mutants were examined for their ability to generate lesions in athymic nude mice. Viral activities were examined by qPCR, immunohistochemistry (IHC), in situ hybridization (ISH), and transmission electron microscopy (TEM) analyses. We demonstrated that viral DNA replication and tumor growth occurred at both cutaneous and mucosal sites infected with each of the mutants. Infections involving L1ATGko-4m, L2ATGko, and L1-L2ATGko mutant genomes generally resulted in smaller tumor sizes compared to infection with the wild type. The L1 protein was absent in L1ATGko-4m and L1-L2ATGko mutant-treated tissues, even though viral transcripts and E4 protein expression were robust. Therefore, L1 is not essential for MmuPV1-induced tumor growth, and this finding parallels our previous observations in the rabbit papillomavirus model. Very few viral particles were detected in L2ATGko mutant-infected tissues. Interestingly, the localization of L1 in lesions induced by L2ATGko was primarily cytoplasmic rather than nuclear. The findings support the hypothesis that the L2 gene influences the expression, location, transport, and assembly of the L1 protein in vivo. L1 and L2, the papillomavirus major and minor capsid proteins, have been studied extensively in vitro . The cotThe papillomavirus community has gained significant understanding of L1 and L2 at the molecular level . Yet, thThe mouse papillomavirus (MmuPV1) was first identified in an athymic nude mouse colony in 2011 . Since tThe functions of MmuPV1 L2 and the interaction between L1 and L2 have not been studied previously . TherefoThe mutant genomes were encapsidated in HPV31 L1/L2 capsids to generate quasiviruses (QVs) using a previously established in vitro production system . OutbredL1 mutants were generated by mutating either the first two methionines (designated as L1ATGko-2m) or all four start codons (designated L1ATGko-4m) from ATG to ACG . The L2 Each mutant genome was released from the vector with BamHI digestion and retrieved with a gel extraction kit  to be used for quasivirus (QV) production as previously described . HPV31 Lnu/nu nude mice (4–5 weeks old) were obtained from ENVIGO and were housed in sterile cages within sterile filter hoods and were fed sterilized food and water in the BL2 animal core facility. Female NU/J heterozygous mice were purchased from the Jackson’s laboratory and maintained in an animal core facility at the University of Manitoba. Mice were sedated i.p. with 0.1 mL/10 g body weight ketamine/xylazine mixture (100 mg/10 mg in10 mls ddH2O). Each nude mouse was infected with one mutant virus at both cutaneous and mucosal sites simultaneously. All animals were treated with Depo-Provera three days before scarification as described previously [TM, and the base of the tongue was wounded with a microneedle as described previously [8 genome DNA equivalent) were applied to each wounded site followed either by scratching with a TB needle for cutaneous sites or a Doctor’s BrushpickTM at the mucosal sites [All mouse work was approved by the Institutional Animal Care and Use Committee of Pennsylvania State University’s College of Medicine and University of Manitoba. Female HSD outbred Foxn1eviously ,23. Muzzeviously ,25. Folleviously ,22. The al sites . To bloc8 copy numbers, . The qPCR reactions were run in an AriaMx cycler (Agilent). Each reaction consisted of 500 nM specific primer pairs, 250 nM double-labeled probe. The following cycling conditions were applied: one cycle of 95 °C for 10 min followed by 40 cycles of 94 °C for 15 s and 60 °C for 1 min [ΔCt) demonstrated the relative viral DNA load in each sample as previously described [DNA was extracted from lavage/swab specimens using the DNeasy kit . Linearized MmuPV1 genome DNA was used for standard curve determination by Probe qPCR analysis . The primer pairs (5′GGTTGCGTCGGAGAACATATAA3′ and 5′CTAAAGCTAACCTGCCACATATC3′) and the probe 6-5′ FAM-TGCCCTTTCA/ZEN/GTGGGTTGAGGACAG 3′-IBFQ 3′) that amplifies a portion of E2 were used . Oral, vor 1 min . All samor 1 min . Fold chescribed .Our unpublished observations have shown that MmuPV1 does not tolerate foreign insertions such as GFP and luciferase. Such genomes reverted to wild type in vivo. To confirm whether infection was initiated and sustained by MmuPV1 L1 or L2ATGko, we amplified the viral genome directly from the DNA extract of lesions or lavages/biopsies using the illustraTempliPhi Amplification Kit . A MmuPV1 specific primer mix covering the entire MmuPV1 genome was used for amplification . The amplified DNA was subsequently sequenced and aligned with the wild-type MmuPV1 using DNAMAN software.Infected tissues were harvested at the termination of the experiments and stored in liquid nitrogen before being used for DNA and protein extraction. For ELISA, infected tissue suspension (2 µg protein/well) was used as an antigen for viral detection, as described previously . A panelFor transmission electron microscopy (TEM), the tissue was immersion-fixed for 24 h in Karnovsky’s fixative  in 0.1 M sodium cacodylate buffer, pH 7.3 . FollowiFrozen tail and vagina tissues were crushed with a hammer while in liquid nitrogen and transferred to immunoprecipitation (IP) buffer  with Protease and Phosphatase Inhibitor Cocktail  added just before use. Protein concentration was determined with a BCA Protein Assay kit . Then, 10 µg vagina positive control and 40 µg L1-L2 ATGko protein (3 separate samples) were run on a Novex WedgeWell 4–20% Tris-Glycine Gel  and transferred to a PVDF membrane . MPV.B9 (1:200) and Anti-mouse IgG1 HRP  were used to probe for L1 in Western blot analysis. An IP/Western was also conducted for the tail tissues. Then, 25 µg positive control and 3 separate L1-L2ATGko samples were incubated overnight at 4 °C with 2 µg MPV.A4 mab with end over end mixing. Then, 20 µL protein A/G agarose beads were added for 1 h followed by 4 washes of the pellets with IP buffer. The samples were run and probed as described above. Protein loading controls were confirmed by Coomassie stain (data not shown).t-test were used by comparing differences between two groups. Differences were significant at p < 0.05.The data were statistically analyzed with one-way ANOVA analysis in Sigmaplot 12 software. A Mann–Whitney rank-sum test and unpaired Student’s According to an in vitro study, the first two methionines that potentially encode the MmuPV1 L1 protein are not necessary for proper VLP production . To testSince mutants with abrogation of the first two methionines displayed an infection pattern similar to that of the wild type, we generated the next L1 mutant with the first four start codons mutated from ATG to ACG and labeled the mutant L1ATGko-4m. The L1ATGko-4m mutant was tested at both cutaneous A and mucThe productive stage of the viral life cycle, in which progeny virus is made, arises within the terminally differentiating compartment of stratified squamous epithelia ,31. To dp > 0.05 unpaired Student’s t-test). This finding suggests that L2 does not play a role in MmuPV1 DNA amplification. However, the presence of only a limited number of viral particles supports the hypothesis that L2 may play a critical role in viral particle assembly and/or transport in vivo.Very few studies have been done to unravel the interactions between L1 and L2 in vivo. In this study we wanted to determine whether the absence of L2 influences the expression of L1 at infected sites. For wild-type virus-induced lesions, full-length L1 was found in the nucleus. An in-house monoclonal antibody MPV.B9 was used for detection  Figure A. HoweveOne of the best-known hallmarks of HPV infection is the induction of DNA synthesis in the suprabasal layers of the epidermis. This reprogramming of suprabasal cells to support DNA synthesis is thought to be required for amplification of the viral DNA that is necessary to produce progeny virus . To deteFive HSD nude female mice were infected at both cutaneous (tail and muzzle) and mucosal  sites with the L1-L2ATGko quasiviruses. Out of five animals in repeated experiments, three animals grew tail lesions, and two animals grew muzzle lesions . CompareA similar pattern was observed at the two mucosal sites (vagina and tongue), but very low to absent viral activity was found in the anal tract A. Viral p > 0.05, Mann–Whitney rank-sum test, vs. negative control). Collectively, these observations support the prediction that four ATG to ACG knockouts in L1 completely abrogated L1 expression in these tissues. We do not currently have the tools to investigate whether a truncated L2 could be expressed by the L2ATGko mutant.To further determine whether viral transcripts were impacted in L1-L2ATGko mutant-induced lesions, we used RNA-ISH to detect L1, L2, and E4 transcripts in these tissues. As shown in p < 0.05 unpaired Student’s t-test). However, there was no difference in DNA copy number between vagina and anus for the L2ATGko infections (p > 0.05 unpaired Student’s t-test). These findings indicate that L1 may contribute to the differential tissue susceptibility of the vaginal and anal tracts.To explore how L1 and L2 influence mucosal infections in the animals, twelve athymic nude mice were divided into four groups (N = 3 mice/group) and each group was infected with wild-type , L1ATGkoIn this study, we characterized the role of the capsid proteins L1 and L2 in the viral life cycle of MmuPV1 using our recently established mouse papillomavirus model, which is a model that displays a broad tissue tropism ,20. We gcytoplasm, suggesting that the absence of L2 has an impact on the location of full-length L1 expression and/or transport in vivo. We postulate that in the absence of L2, the nuclear localization signal at the C terminus of full-length L1 is disactivated, whereas in the presence of L2, the influence of L2 protein predominates.The double L1-L2ATGko mutant failed to induce tail lesions in two of the five tested mice, and the lesions were significantly smaller than those induced by L1ATGko or L2ATGko mutants, suggesting that the mutations in this genome influenced the infectivity of the mutants. Full-length L1 expression was abrogated in the L1ATGko-4m and L1-L2ATGko-infected tissues as determined by probing with a panel of anti-L1 antibodies including MPV.A4 and MPV.B9, but viral DNA replication, viral transcripts of E4, L1, and L2, as well as E4 expression were not compromised. No L1 signals in L1-L2ATGko mutant-infected tissues were found using the antibody against group-specific antigen (GSA), which targets the carboxy-terminus of L1 . InteresThe L1 and L2 proteins of the cottontail rabbit papillomavirus have been found to be non-essential for tumor outgrowth in the rabbit papillomavirus model ,6,7,8,9.DNA replication in lesions induced by the mutant genomes was comparable to that of wild-type lesions. This suggests that neither L1 nor L2 protein is required for viral replication. On the other hand, localization of the full-length L1 protein was dependent upon the expression of L2. For example, the mutation of the L2 ATG resulted in the relocation of full-length L1 expression to the cytoplasm. Our work has been hampered by the lack of an antibody to L2, and studies are ongoing to further elucidate the nature of L1/L2 interactions. The findings presented here support earlier in vitro findings that the likely start codon for L1 is the 3rd methionine at amino acid position 28 . The L1AIn conclusion, we have demonstrated that both papillomavirus capsid proteins (L1 and L2) are dispensable for viral DNA replication and tumor growth in vivo. However, L1 and L2 do appear to contribute to viral production and differential tissue susceptibility.


To this aim, the influence of incorporation of graphene nanosheets into ultra-high molecular weight polyethylene (UHMWPE) on the thermal behavior and degradation kinetics of UHMWPE/graphene nanocomposites was investigated. Scanning electron microscopy (SEM) analysis revealed that graphene nanosheets were uniformly spread throughout the UHMWPE’s molecular chains. X-Ray Diffraction (XRD) data posited that the morphology of dispersed graphene sheets in UHMWPE was exfoliated. Non-isothermal differential scanning calorimetry (DSC) studies identified a more pronounced increase in melting temperatures and latent heat of fusions in nanocomposites compared to UHMWPE at lower concentrations of graphene. Thermogravimetric analysis (TGA) and derivative thermogravimetric (DTG) revealed that UHMWPE’s thermal stability has been improved via incorporating graphene nanosheets. Further, degradation kinetics of neat polymer and nanocomposites have been modeled using equations such as Friedman, Ozawa–Flynn–Wall (OFW), Kissinger, and Augis and Bennett’s. The "Model-Fitting Method” showed that the auto-catalytic nth-order mechanism provided a highly consistent and appropriate fit to describe the degradation mechanism of UHMWPE and its graphene nanocomposites. In addition, the calculated activation energy (E Ultra-high molecular weight polyethylene (UHMWPE) is renowned for its stellar mechanical properties, high abrasion resistance, low moisture absorption, low friction coefficient, and excellent chemical stability ,2. UHMWPThe final properties of nanocomposites generally depended on the nature of the nanofillers and how they were fabricated. The main problem with the implementation of melt intercalation and solution casting methods for UHMWPE composite preparation is its extra high viscosity and lack of flowability in the melted UHMWPE and its solution. The in situ polymerization method is, by far, the best method for dispersing fillers for more efficient contact between the filler and the polymer matrix ,11. The 4) and sulfuric acid (H2SO4), while in the Brodie method, potassium chlorate (KClO3), and nitric acid (HNO3) are used to oxidize graphite. Staudenmaier’s method also uses a procedure like Brodie’s to oxidize graphite to GO. Stürzel et al.  vs. (1/T) is a straight line at several heating rates, and the Ea is measured from the slope. The equation used in the OFW method was derived from the assumption of constant Ea, and an error can be calculated by comparison with the Friedman results by incorporating a systematic error in the measurement of Ea as E varies with α  vs. 1/Tm plot, Ea can be achieved. Like the Kissinger method, the Ea values determined from Augis and Bennett method are related to the peak temperature of the DTG curve. These values are equal to the values of the other isoconversion methods and are listed in Where Ta, it can be observed that all five methods used in this study showed consistent values of Ea for neat UHMWPE and the samples containing 0.9 and 2.1 wt.% of graphene. However, for the sample containing 3.4 wt.% of graphene, the Ea values determined by Friedman and OFW methods were significantly different from the results obtained by other methods. This may be due to some existing uncertainties over baselines of the thermal analysis data or limited accuracy of determination of transformation rates [a. In addition, the obtaining results from calculation of Ea using different methods confirmed this fact that the presence of graphene and its interactions with the UHMWPE matrix generally increase the thermal stability and Ea of the polymer. However, above a specific level of graphene contents in UHMWPE matrix that the activation energy decreased is most probably due to the destructive effect of active free radicals and agglomeration effects. The study of kinetic modeling and thermal degradation behavior of polymers including UHMWPE helps with better understanding of predicting thermal degradation to prevent the weaknesses of the polymeric products. As UHMWPE is an engineering polymer which is used in different industries including aerospace, the information about its thermal behavior is crucial. In addition, the kinetic studies provide information to develop pyrolysis reactors used for thermal modification of polymeric wastes . By compon rates . Althougon rates . Accordia in all the methods showed a stabilizing effect on the degradation of the polymeric matrix as the graphene loading increased up to 2.1 wt.%. However, the graphene content of 3.4 wt.% or more decreased the Ea slightly. Modeling of degradation kinetics of UHMWPE and its nanocomposites were conducted using different methods, and the results demonstrated that although the auto-catalytic nth-order mechanism was not able to be fitted accurately with the actual TGA curves, this model could describe the degradation mechanism of UHMWPE and its nanocomposites by accurately showing the order of magnitudes of temperature thermogram peaks as well as by predicting activation energies consistent with model-free methods such as Kissinger. Furthermore, all modeling approaches in this study estimated similar values for Ea except the values obtained by Friedman and OFW methods for UHMWPE/graphene 3.4 wt.%, which were not close to the values obtained by other methods, which can be due to some existing uncertainties over baselines of the thermal analysis data or limited accuracy of determination of transformation rates. Thermal degradation behavior of UHMWPE/graphene nanocomposites was evaluated with different graphene contents . The prepared nanocomposite was characterized by SEM and XRD, and its thermal properties were investigated by TGA, DSC, and DTG. It is concluded from the TGA results that the thermal stability of these nanocomposites increased up to 370 °C without any significant mass loss via incorporating graphene nanosheets. Thermal degradation kinetics were investigated using model-free and model-fitting methods. The calculated E


In this work, we present the fabrication and characterization of bulk-heterojunction solar cells on monolayer graphene (MLG) with nickel-grids (Ni-grid) as semitransparent conductive electrode. The electrodes showed a maximum transmittance of 90%  and a sheet resistance down to 35 Ω/□. On these new anodes, we fabricated TCO free BHJ-SCs using PTB7 blended with PC70BM fullerene derivative as active layer. The best device exhibited a power conversion efficiency (PCE) of 4.2% in direct configuration and 3.6% in inverted configuration. The reference solar cell, realized on the ITO glass substrate, achieved a PCE of 6.1% and 6.7% in direct and inverted configuration respectively; for comparison we also tested OSCs only with simple Ni-grid as semitransparent and conductive electrode, obtaining a low PCE of 0.7%. The proposed approach to realize graphene-based electrodes could be a possible route to reduce the overall impact of the sheet resistance of this type of electrodes allowing their use in several optoelectronic devices. The act2 connected with a sourcemeter (Keithley 2420); the irradiation level was verified at the same height and position where the solar cell is placed with a calibrated pyranometer (Skye SKS1110). The samples were covered during the measurement with a shadow mask with an opening area of 0.1 cm2 to illuminate only one cell at a time. A customized tool comprising a sourcemeter (Keithley 2612) and a monochromator  was used to measure the incident-photon-to-current conversion efficiency (IPCE) values of the device.Device performance was evaluated under a Class A solar simulator  at AM 1.5 G and 100 mW/cmZo is the free space impedance (377 Ω). Higher values of δdc/δop correspond to enhanced electro-optical performance.Aiming at choosing the best grid geometry for the realization of the semitransparent MLG/grid electrode, we tested several grid square dimensions evaluating the transmittance and sheet resistance. The results are reported in The best balance between transmittance and sheet resistance was found for a 75 µm × 75 µm square dimension showing the 79.8% of transmittance and 165 Ω/□ of sheet resistance and for 150 µm × 150 µm square dimension with 5 µm of width, showing the 86% of transmittance and 355 Ω/□ of sheet resistance.The optimum grid was then used for the transferring of the graphene layer. The optimized graphene/Ni-grid geometry was then used as an electrode in OSCs: three different thicknesses of the grids, 20 nm, 40 nm and 80 nm were tested for the direct architecture, while the 40 nm thick grid, which was giving the best performances on the direct architecture, was chosen for the inverted one. The produced devices were compared with identical ones fabricated on glass/ITO, on bare Ni grids on quartz and monolayer graphene (MLG) on glass for both architectures. The measures on the MLG/Ni-grids have been performed on eight samples of each type, the resulting statistics are reported in The best results for the direct architecture as function of the different anodes and thicknesses of the grid are reported in 2, Voc = 633 mV, FF = 40.2% and PCE = 0.5%) and only graphene  were lower than the most efficient Ni-grid/MLG based device . The primary differences were in short circuit current (Jsc) and fill factor (FF); the decrease in Jsc reflected the lower collection charge of the Ni-grid due to the absence of a continuous anode, while, in the case of the MLG graphene, it could be ascribed to the defects in the graphene layer induced by the transferring method [As expected, the performance of the devices realized on Ni-grid on quartz-based devices , showed a reduction in efficiency of about 30% when compared with the ITO-based devices . 2 and 10.8 mA/cm2, respectively, on ITO/glass and MLG/Ni-grid/quartz anodes.In the case of direct architecture, the best performance of the solar cells obtained for MLG/Ni-grid (40 nm)-based devices  and a sheet resistance down to 35 Ω/□. The hybrid transparent Ni-grid/MLG films were then employed as semitransparent devices for the realization of PTB7:PCBM organic solar cells, yielding a maximum PCE of 4.2% in direct architecture. The OSCs with the hybrid anodes exhibited significantly better performance in comparison with the ones realized with only MLG and raw Ni-grid. However, when compared with the reference cells based on ITO they demonstrated a 30% and 46% reduction in efficiency with a direct and inverted architecture respectively. These results can be ascribed mainly to the higher sheet resistance of the Ni-grid/MLG electrodes that cause a decrease in the FF of the solar cells and also to the defects at the interfaces between the semitransparent electrodes and the solar cells layers. Further improvements in terms of efficiency of the devices could be achieved reducing the contamination of the graphene layer during the transfer on the grids reducing the effects of the parasitic resistances of the devices, in addition to the introduction of proper doping of the graphene layer. Considering the realization of organic solar modules, the introduction of the grids would be of paramount importance when aiming to scale up to large area devices. Their geometry will be of fundamental importance to guarantee uniform deposition of the different layers, allowing one to obtain working devices and representing a possible route towards the realization of a more sustainable, ITO-free organic solar cells and modules.


The obtained results reveal the importance of defects in graphene structure, which play a key role in wetting mechanism and the formation of AgCu alloy. As a consequence, we observe a change of the wetting behavior and topology of both bulk and adsorbed Ag atoms by using Voronoi analysis (VA). Despite the differences in time scale, atomistic simulations allowed us to catch the early stages of wetting, which are important for explaining the final stage of wetting delivered from experiment. Our findings reveal also graphene translucency to metal-metal interactions, observed in previous papers.Understanding the atomic-level phenomena occurring upon the wetting of graphene-coated Cu with liquid Ag is pivotal for the description of the wetting phenomenon and the role of graphene as a diffusion barrier. We have performed molecular dynamics (MD) simulations and confronted with our present experimental results to characterize wetting behavior of graphene coated Cu surfaces. Perfect and defected graphene layers covering Cu surface were wetted with liquid Ag droplet at 1273 K. Structural and topological aspects are discussed to characterize structure of the liquid Ag droplet and a product of wetting reaction occurring on Cu/Gn and Cu/Gn Graphene is known from its extraordinary properties, which have allowed it to find a broad spectrum of applications ranging from electronics to medicine as well as gain significant research interests . Potential applications include sensors, capacitors, energy storage devices, solar cells, transparent conductive coatings, and tissue engineering applications ,2,3. GraDiffusion at the solid/liquid interface plays a huge role in electronic packaging industry, where tin-silver-copper (SAC) solders are mostly used as joints. They form intermetallic compounds (IMCs) with substrates like Cu or Ni. The extensive growth of IMCs is unwanted due to worsening joint properties and enhancing the risk of fracture, which is dangerous for electronic devices ,13. As yIn this context, there is a gap in the knowledge about the structure and properties of metal/graphene interfaces as well as wetting of graphene-coated metallic substrates with a metal droplet. So far, few papers exist which provide knowledge about graphene layer and metal/graphene interface wetted with water requiring a further explanation ,15,16. LAtomic-scale physical models, such as molecular dynamics and Monte Carlo methods, offer a possibility of gaining insight into the wetting phenomenon, interface structure, and kinetics of potential reactions on atomic level. They involve physical and chemical equations for describing interatomic interactions, which are then pivotal for a proper description of the wetting reaction on metal and metal/graphene substrates. Only a few computational papers can be found for graphene-coated substrates as compared to a graphene layer wetted by liquid metal droplets. Caccia et al. performed ab initio calculations for a study of wetting translucency in Au droplet on a pure and graphene-supported 4H-SiC substrate as well as graphene substrate with variable layer number . As a reIn this work, we provide the first atomic level insight into a wetting mechanism, structure, and topology of a silver droplet wetting graphene-coated Cu substrate at 1273 K. The aim of our paper is to unveil the mechanism of graphene wetting with liquid Ag droplet, and the structure of both Cu/graphene interface and the liquid droplet. For comparison, pure Cu substrate and a graphene layer are also considered. Here we present molecular dynamics simulation results and experimental observations. To get a more detailed insight into the mechanism of graphene wetting and its better understanding, we monitored changes of contact angle with time and provided information on structural and topological aspects to better understand the behavior of liquid droplet and the wetting phenomenon. def). In the last case, two atomic columns in the center of the graphene layer were removed. Depending on the interaction type, we used charge-optimized many body potential (COMB3) formalism [Molecular dynamics simulations have been performed for the study of structure, wetting properties and spreading mechanism of silver droplet on Cu/graphene substrate at 1273 K. We have considered four different substrate variants wetted with a silver droplet. They are pure Cu, graphene layer (Gn), Cu substrate covered by both a perfect graphene (Cu/Gn) and defect containing graphene layers  potential for describing the van der Waals interaction between liquid Ag droplet and graphene layer:For describing metal-metal interaction (Ag-Ag and Cu-Ag), the embedded atom model (EAM) potential was used , with paAg-Cε) and interatomic distance (Ag-Cσ) between Ag and C were 0.0301 eV and 3.006 Å, respectively, as standard numerical values reported in the literature [The interaction energy  until a droplet was formed. In the second step, the Ag droplet was placed about 7 Å over the four different substrate variants. MD simulations with canonical ensemble (NVT) were performed at 1273 K for three substrate variants with maximum simulation time up to 0.5 ns, and 1 ns for Cu/Gndef/Ag system. In the case of variant of graphene substrate with Ag droplet, rigid body assumption was made for graphene layer during whole simulation time. Nose-Hoover thermostat, implemented in LAMMPS code, was used for maintaining a target temperature during whole simulation. The visualization of wetting phenomenon occurring for four substrates variants was made by using Visual Molecular Dynamics (VMD) package [In the first step of our MD simulation procedure, both Cu matrix and graphene (Gn) layer were separately heated from 0 K to 1273 K before wetting studies, and then both were thermalized during 20,000 ps with 0.5 fs time step. Cu matrix with approximately 100 Å × 100 Å × 30 Å dimensions was tightly covered by a graphene layer of similar dimensions in the xy plane on which periodic boundary conditions were imposed. The Cu substrate contained 20,000 atoms, the Gn layer 7500 atoms, the Gn package .Selected atomic structures of each considered substrate variant wetted with Ag droplet were minimized using conjugated gradient method, as implemented in LAMMPS, before performing structural and topological analyses as well as calculating wettability. Time dependence of contact angles (CA) was computed for four configuration variants used to describe wetting properties of Cu matrix covered by a graphene and a single graphene substrate. Spreading area and the height of droplet on a substrate were also determined using MD simulation derived structures and compared with experimental data. Procedure of CA determination is described in the modelling section of the n3, n4, n5, n6, n7) is used to differentiate the types of VC, where ni is the number of i-edged sides in the polyhedron. This tool allows to recognize body- and face-centered cubic structures (bcc and fcc), as well as hexagonal close-packing (h.c.p).Voronoi analysis is based on three dimensional Voronoi tessellation, which provides a detailed description of atom linkage in 3-dimensional space. It gives an deep insight into local atom changes through a complete geometrical construction of atom to its neighbor atoms. More details about VA can be found in papers ,45,46. VavP, and its coefficient pK, according to a formulation proposed by Pstruś et al. [P0 was needed, which is a projection of a sphere on the surface, to compute pK coefficient. We considered atomic structure of Ag droplet in Gn/Ag variant at 1 ps, and then computed the mean value of sphere area by using the formula: pK coefficient as follows:Wettability was measured by determining spreading area ś et al. . For thihK coefficient, where the radius of the sphere was already known (D0). The height of the droplets on considered variants were calculated based on maximum and minimum value of Z for Ag atoms of geometries at 1200 ps. hK coefficient was computed using formula:Similar methodology was used to calculate pK coefficient.A Chemical Vapour Deposition method was used for graphene growth on Cu foil with the dimensions of 30 × 30 × 0.25 mm and its purity of 99.999% . We followed step-by-step a procedure of graphene deposition as it is described by Pstruś et al. . Ag sampdef) is presented in def substrates.Molecular dynamics simulations results obtained at 1273 K for Ag droplet wetting graphene-coated Cu substrates are discussed below. For clarity of the presentation, we present MD results for Ag droplet wetting a graphene layer and a pure Cu substrate in In case of perfect graphene-coated Cu substrate, silver droplet slowly spreads on Cu/Gn surface and its shape also slowly changes, which is associated with simultaneous diffusion of silver atoms on the top surface of Ag drop. Such behavior illustrates a non-reactive wetting for which a stronger interaction between droplet atoms dominate over the weaker ones between Ag and C. Similar spreading mechanism occurs for Ag wetting a perfect graphene layer as presented in def substrate wetted with liquid silver differs from the simple case described above. A mixture of non-reactive wetting and a reactive wetting is observed. To get deeper insight into these phenomena, we present in def wetting with liquid Ag droplet by combining atomistic simulations with experiment. At 1st stage, liquid Ag droplet is approaching Cu/Gndef substrate and Ag atoms are getting to interact with Cu atoms through the defect. Simultaneously, an impairment of graphene layers between part A and part B, and surface diffusion of Ag atoms on graphene layer occur at 2nd stage. The bottom part of def interface and then move on Cu substrate before its dissolution. Consequently, the Cu-graphene distance increases and graphene layer  climbs al stage . The plaDue to the dissolution of carbon in silver, based on the phase diagram for Ag-C  and defedef/Ag systems, we computed radial distribution functions (gij(r)) for each respective atomic pair contributing to the considered substrate variants with Ag droplet. The pair distribution function of the Ag-Cu pair is presented in ij(r) are illustrated in Ag-Cu(r) for Cu/Gndef/Ag system exhibits three peaks of relatively low intensity as compared to that computed for Ag-Cu pair of AgCu alloy formed upon reactive wetting of a pure Cu substrate  for the latter systems was rescaled by 0.1 to be able to make a reliable comparison because of high intensity of the first pronounced peak of gCu-C(r) for Cu/Ag system. Two features require more explanation, namely the positions of the first and third peak of gAg-Cu(r). The first peak of gAg-Cu(r) allocates around 2.9 Å similarly to the position of the first pronounced peak of gAg-Cu(r) for the AgCu alloy  in crystalline-to-amorphous transformation of the Cu substrate as showed snapshot of spreading evolution for Cu/Ag systems (see Cu-Cu(r) drawn in For a deeper insight into the impact of wetting on the structure of Cu/Gn/Ag and Cu/Gntems see  and gCu-def/Ag systems, we have also inspected time evolution of Cu-C bond length. Due to the gap dividing the “defected” graphene layer (Gndef) into two parts , whose abundance is drawn in def substrate. Moreover, 5 digit VC indices are also common for both Gn and Cu/Gndef, which are of similar abundance to VC indices of 1st group. When analyzing VC types present in one substrate variant presented in def/Ag. There is no VC types which would occur only for the Cu/Gn substrate. An abundance graph of VC types detected for Ag atoms forming an AgCu alloy in place of the structural defect in the graphene layer deposited on Cu substrate is presented in def substrate leading to alloy formation.Looking deeper into the wetting phenomenon, we performed a Voronoi analysis (VA) to characterize the topology of Ag atoms adsorbed on two considered graphene-coated Cu substrates and of Ag atoms in the bulk liquid phase. Abundance graphs of Voronoi cell (VC) types for Ag atoms being in contact with the graphene layer is presented in def substrate to those wetting Gn layer, which could not have been caught by visual perception and even from chemical analysis (In a view of the performed Voronoi analysis for Ag atoms adsorbed on a graphene layer, we can observe a similarity in behavior of Ag atoms adsorbed on Cu/Gnanalysis . This redef substrates at 450 ps is drawn in Abundance of 15 most representative VC topologies for bulk liquid Ag droplets wetting Cu/Gn and Cu/GnWe have performed molecular dynamics simulations to unveil the mechanism of liquid Ag wetting Cu substrate covered with perfect graphene layer and defect containing one at 1273 K, which is supported with our present experimental result. We have also discussed structural and topological aspects to correctly describe the impact of wetting phenomenon, occurring on different types of substrates. The results clearly show, that the presence of defect in graphene structure affects wettability behavior, and is a pivotal for the occurrence of diffusion as well. Moreover, introducing graphene at the Cu/Ag interface, allows to observe graphene translucency to metal-metal interactions.Our work shows, that with molecular dynamics we were able to simulate the early stages of wetting of Cu/Gn structure with a silver droplet, observed experimentally. It was achieved mainly due to implementing a defect in graphene in the simulations. Combination of those two approaches allows us to propose a mechanism of wetting, as well as graphene detachment from the substrate, resulting in slowing down the diffusion process in the system. However, several aspects still remain to be considered for a complete description of the wetting process, such as longer simulation time parallel with shortening of the wetting time in the experimental studies. They also concern a consideration of more complex graphene models to MD simulations, including the experimentally-observed defects for CVD-grown graphene layers. Nevertheless, despite the rather simple simulation setup, we were able to obtain consistent results.