ABSTRACT
OBJECTIVE: To determine the preventive effect of coenzyme Q10 (CoQ10) on the testicular histology of rats exposed chronically to mosquito coil smoke. STUDY DESIGN: Experimental study. Place and Duration of the Study: Department of Anatomy, Army Medical College/National University of Medical Sciences, Rawalpindi, Pakistan, from January to December 2020. METHODOLOGY: Thirty male Sprague Dawley rats were divided into three groups of 10 rats each. Group A was the healthy control. Group B rats were exposed to allethrin-based mosquito coil smoke for 12 weeks (4 hours/day). Group C rats received coenzyme Q10 (CoQ10, 10mg/kg/day) through oral gavage, in addition to 12 weeks of mosquito coil smoke exposure (4 hours/day). At the end of the study, testicular histology was compared among three groups including the germinal epithelium height, seminiferous tubule diameter, and testicular capsule thickness, while adjusting for the body weight variations among rats. RESULTS: The rats in Group B, exposed only to mosquito coil smoke showed testicular disruption, characterised by dilated seminiferous tubules (p <0.001), reduced germinal epithelial height (p <0.001), and thickened testicular capsule (p <0.007), as compared to the control group rats. However, the germinal epithelium height (p = 0.73) and testicular capsule thickness (p = 0.31) of rats receiving CoQ10 in addition to mosquito coil smoke inhalation were not significantly different from the control group. CONCLUSION: Prolonged inhalation of allethrin-based mosquito coil smoke can cause testicular disruption among rats. The oral CoQ10 administration can effectively prevent the histomorphological adverse effects on the testis among rats exposed to mosquito coil smoke. KEY WORDS: Allethrin, Coenzyme Q10, Germinal epithelium, Mosquito coil, Seminiferous tubules, Testicular capsule.
Subject(s)
Rats, Sprague-Dawley , Testis , Ubiquinone , Animals , Male , Ubiquinone/analogs & derivatives , Ubiquinone/pharmacology , Ubiquinone/administration & dosage , Rats , Testis/drug effects , Testis/pathology , Smoke/adverse effects , Allethrins/pharmacology , Smoke Inhalation Injury/prevention & control , Smoke Inhalation Injury/pathologyABSTRACT
The Towards Gender Harmony (TGH) project began in September 2018 with over 160 scholars who formed an international consortium to collect data from 62 countries across six continents. Our overarching goal was to analyze contemporary perceptions of masculinity and femininity using quantitative and qualitative methods, marking a groundbreaking effort in social science research. The data collection took place between January 2018 and February 2020, and involved undergraduate students who completed a series of randomized scales and the data was collected through the SurveyMonkey or Qualtrics platforms, with paper surveys being used in rare cases. All the measures used in the project were translated into 22 languages. The dataset contains 33,313 observations and 286 variables, including contemporary measures of gendered self-views, attitudes, and stereotypes, as well as relevant demographic data. The TGH dataset, linked with accessible country-level data, provides valuable insights into the dynamics of gender relations worldwide, allowing for multilevel analyses and examination of how gendered self-views and attitudes are linked to behavioral intentions and demographic variables.
Subject(s)
Femininity , Masculinity , Stereotyping , Female , Humans , Male , Attitude , Students , Surveys and Questionnaires , Gender Role , Self ConceptABSTRACT
As of 2021, over 5.4 million Venezuelans have fled their home country in search of safety, food, medicine and access to essential services. This is the most substantial exodus in the recent history of Latin America. Colombia has received 2 million of these refugees, making it the nation host to the greatest number of Venezuelan refugees. The present research aims to examine the relations between the sociocultural and psychological factors that are associated with Psychological Adaptation of Venezuelan refugees living in Colombia. We also examined how these relations were mediated by the acculturation orientations. Among Venezuelan refugees, higher Psychological Strength, lower Perceived Discrimination, higher National Identity and higher Outgroup Social Support, were significantly associated with higher engagement with Colombian society and better Psychological Adaptation. Orientation to the host (Colombian) society mediated the association between (a) National Identity and Psychological Adaptation, (b) Outgroup Social Support and Psychological Adaptation and (c) Perceived Discrimination and Psychological Adaptation. The results may inform refugee receiving societies of some essential factors and positive strategies behind adaptation of refugees.
Subject(s)
Acculturation , Refugees , Humans , Refugees/psychology , Colombia , Emotional Adjustment , Adaptation, PsychologicalABSTRACT
ABSTRACT The tendency to be permissive in face of a discriminatory situation is called collusion. The present study aimed to define and characterize collusion, and identify the variables connected to it, considering the perspective of different identity groups. Participants were 31 individuals divided in seven focus groups. The analysis indicated four categories connected to collusion: a) Close Relations: valuable interactions through which individuals learn behavioral patterns that lead to permissiveness; b) Group Identity: social identities, and intergroup relations patterns; c) Situation: characteristics of the situation in which discrimination is observed; and d) Cost-Effective Balance: perception the individuals have regarding the impact of their actions on the context and the cost attached to it.
RESUMO A tendência das pessoas de serem permissivas diante de uma situação discriminatória é chamada colusão. O presente estudo teve como objetivo definir e caracterizar colusão e identificar as variáveis a ela associadas, a partir da perspectiva de diferentes grupos identitários. Os participantes foram 31 sujeitos divididos em sete grupos focais. A análise indicou quatro categorias ligadas à colusão: a) Relações Próximas: interações por meio das quais os indivíduos aprendem padrões de comportamento que levam à permissividade; b) Identidade Grupal: identidades sociais e padrões de relações intergrupais; c) Situação: características da situação em que se observa discriminação; e d) Relação Custo-benefício: percepção que os indivíduos têm sobre o impacto de suas ações no contexto e o custo que lhes está associado.
ABSTRACT
Based on acculturation psychology and intergroup emotions theory, the current experimental study assessed the effects of Muslims' perceived acculturation strategies by the majority group on social exclusion of Muslims in Canada, and to what extent religious resentment mediated the relationship between Muslims' perceived acculturation strategies and social exclusion. The experimental study used a vignette-based approach. This model was examined among 190 non-Muslim Canadians. Results showed that when Muslims were viewed as assimilated in Canadian society, social exclusion of Muslims and religious resentment toward Muslims decreased. Furthermore, religious resentment mediated the association between Muslims' perceived acculturation strategies and social exclusion only when Muslims were perceived as assimilated. Our findings suggest that Canadian majority-group members indicated positive attitude toward Muslims when they were identified as assimilated in Canadian society. Results are discussed in terms of implications for future studies and intergroup relations.
ABSTRACT
In recent years, CRISPR-Cas (stands for: clustered regularly interspaced short palindromic repeats - CRISPR associated protein) based technologies have gained increasing attention in the biosensing field. Thanks to excellent sequence specificity, their use is of particular interest for detecting nucleic acid (NA) targets. In this context, signal generation and amplification can be realized by employing the cis-cleavage activity of the Cas9 protein, although other options involving the catalytically inactive dead Cas9 (dCas9) are increasingly explored. The latter are however mostly based on complex protein engineering processes and often lack efficient signal amplification. Here we showed for the first time that flexible signal generation and amplification properties can be integrated into the CRISPR-dCas9 complex based on a straightforward incorporation of a DNA sequence into the trans-activating CRISPR RNA (tracrRNA). The intrinsic nuclease activity of the engineered complex remained conserved, while the incorporated DNA stretch enabled two modes of amplified fluorescent signal generation: (1) as an RNA-cleaving DNA-based enzyme (DNAzyme) or (2) as hybridization site for biotinylated DNA probes, allowing subsequent enzyme labeling. Both signal generation strategies were demonstrated in solution as well as while coupled to a solid surface. Finally, in a proof of concept bioassay, we demonstrated the successful detection of single stranded DNA on magnetic microbeads using the engineered CRISPR-dCas9 complex. Thanks to the flexibility of incorporating different NA-based signal generation and amplification strategies, this novel NA engineering approach holds enormous promise for many new CRISPR-based biosensing applications.
Subject(s)
Biosensing Techniques , DNA, Catalytic , CRISPR-Cas Systems/genetics , DNA , DNA, Catalytic/genetics , RNAABSTRACT
Disease treatment with advanced biological therapies such as adalimumab (ADM), although largely beneficial, is still costly and suffers from loss of response. To tackle these aspects, therapeutic drug monitoring (TDM) is proposed to improve treatment dosing and efficacy, but is often associated with long sampling-to-result workflows. Here, we present an in-house constructed ADM-sensor, allowing TDM of ADM at the doctor's office. This biosensor brings fiber optic surface plasmon resonance (FO-SPR), combined with self-powered microfluidics, to a point of care (POC) setting for the first time. After developing a rapid FO-SPR sandwich bioassay for ADM detection on a commercial FO-SPR device, this bioassay was implemented on the fully-integrated ADM-sensor. For the latter, we combined (I) a gold coated fiber optic (FO) probe for bioassay implementation and (II) an FO-SPR readout system with (III) the self-powered iSIMPLE microfluidic technology empowering plasma sample and reagent mixing on the-cartridge as well as connection to the FO-SPR readout system. With a calculated limit of detection (LOD) of 0.35 µg/mL in undiluted plasma, and a total time-to-result (TTR) within 12 min, this innovative biosensor demonstrated a comparable performance to existing POC biosensors for ADM quantification in patient plasma samples, while requiring only 1 µL of plasma. Whereas this study demonstrates great potential for FO-SPR biosensing at the POC using ADM as a model case, it also shows huge potential for bedside TDM of other drugs (e.g. other immunosuppressants, anti-epileptics and antibiotics), as the bioassay is highly amenable to adaptation.
Subject(s)
Biosensing Techniques , Surface Plasmon Resonance , Adalimumab , Drug Monitoring , Fiber Optic Technology , Humans , Microfluidics , Point-of-Care SystemsABSTRACT
Although there is a strong body of literature on the psychosocial distress of refugees from Global South to Global North, there is limited literature on refugees who migrate from Global South (i.e., Syria) to Global South (i.e., Brazil). The present study aims to investigate Syrian refugees in Brazil. Participants were 202 Syrians. Most respondents were men (62.9%), with an average age of 35.9 years old (SD = 11.13). Results suggest an effect of positive ethnic identity in psychological and physical distress, who also perceive more discrimination than individuals who negatively identify with their ethnicity. Structural equation model suggests the fit of the proposed mediation model. Our findings add to the much-needed line of investigations examining Syrian refugees settling in Brazil.
Subject(s)
Refugees , Adult , Brazil , Female , Humans , Male , Refugees/psychology , SyriaABSTRACT
Adult-onset Still's disease (AOSD), a rare systemic inflammatory disorder of unknown etiology, is considered in broad differential in patients with fever of unknown origin or unexplained lymphadenopathy. It is characterized by spiking fever, evanescent salmon-colored maculopapular rash, arthritis or arthralgia, and leukocytosis. Due to broad differentials and lack of any specific diagnostic tests, diagnosis of AOSD poses a great challenge. A concerned physician should have a high index of suspicion while dealing with patients presenting with clinical symptoms of this systemic disorder. We report a case of a 25-year-old African American female with the past medical history of AOSD, who presented with four weeks history of extensive cervical and axillary lymphadenopathy mimicking lymphoproliferative malignancy. Cases have been reported with the development of malignant lymphoma during the course of AOSD. Therefore, careful monitoring of patients with regular follow-up is vital as these patients may develop lymphoproliferative malignancy in the future.
ABSTRACT
Testing multiple biomarkers, as opposed to one, has become a preferred approach for diagnosing many heterogeneous diseases, such as cancer and infectious diseases. However, numerous technologies, including gold standard ELISA and PCR, can detect only one type of biomarker, either protein or nucleic acid (NA), respectively. In this work, we report for the first time simultaneous detection of proteins and NAs in the same solution, using solely functional NA (FNA) molecules. In particular, we combined the thrombin binding aptamer (TBA) and the 10-23 RNA-cleaving DNA enzyme (DNAzyme) in a single aptazyme molecule (Aptazyme1.15-3'), followed by extensive optimization of buffer composition, sequences and component ratios, to establish a competitive bioassay. Subsequently, to establish a multiplex bioassay, we designed a new aptazyme (Aptazyme2.20-5') by replacing the target recognition and substrate sequences within Aptazyme1.15-3'. This designing process included an in silico study, revealing the impact of the target recognition sequence on the aptazyme secondary structure and its catalytic activity. After proving the functionality of the new aptazyme in a singleplex bioassay, we demonstrated the capability of the two aptazymes to simultaneously detect thrombin and NA target, or two NA targets in a multiplex bioassay. High specificity in target detection was achieved with the limits of detection in the low nanomolar range, comparable to the singleplex bioassays. The presented results deepen the barely explored features of FNA for diagnosing multiple targets of different origins, adding an extra functionality to their catalogue.
Subject(s)
Biological Assay/methods , Biosensing Techniques/methods , DNA, Catalytic/metabolism , DNA/chemistry , Nucleic Acids/chemistry , Thrombin/chemistry , DNA, Catalytic/chemistry , Humans , Nanotechnology , Reproducibility of ResultsABSTRACT
The polymerase chain reaction (PCR) has been the gold standard molecular analysis technique for decades and has seen quite some evolution in terms of reaction components, methodology, and readout mechanisms. Nucleic acid enzymes (NAzymes) have been used to further exploit the applications of PCR, but so far the work was limited to the colorimetric G-quadruplex or fluorescent substrate cleaving NAzymes. In this study, a solid-phase, fiber optic surface plasmon resonance (FO-SPR) technique is presented as an alternative readout for PCR utilizing NAzymes. First, the surface cleavage activity of DNAzyme-extended amplicons (DNAzyme-amps) is established, followed by optimization of the PCR conditions, which are required for compatibility with the FO-SPR system. Next, by integrating the complement of a 10-23 DNAzyme into the primer pair, PCR-amplified DNAzyme-amps were generated, tested, and validated on qPCR for the detection of the antimicrobial resistance gene MCR-2. Once validated, this primer concept was developed as a one-step assay, driven by PCR-amplified DNAzymes, for FO-SPR-based sensitive and specific detection. Using gold nanoparticle labeled RNA-DNA hybrid strands as substrate for the DNAzyme, PCR-amplified DNAzyme-amps generated in the presence of MCR-2 gene were monitored in real-time, which resulted in an experimental limit of detection of 4 × 105 copy numbers or 6.6 fM. In addition, the DNAzyme-based FO-PCR assay was able to discriminate between the MCR-1 and MCR-2 genes, to further prove the specificity of this assay. Henceforth, this DNAzyme-based fiber optic PCR assay provides a universally applicable, real-time system for the detection of virtually any target NA, in a specific and sensitive manner.
Subject(s)
DNA, Catalytic/genetics , DNA, Catalytic/metabolism , Membrane Proteins/genetics , Optical Fibers , Polymerase Chain Reaction , Surface Plasmon Resonance/instrumentation , Calibration , Time FactorsABSTRACT
Nucleic acid enzymes (NAzymes) are nucleic acid molecules with catalytic activity. A subset, the RNA-cleaving NAzyme, is characterized by its substrate of choice: an RNA unit. These enzymes have been used for diverse applications, including biosensor development, akin to their protein counterparts. Owing to their function as both biorecognition elements and signal generators, robust bioassays based entirely on NAzyme molecules have been developed. Additionally, unique mechanisms for integration with other biorecognition elements and signal generation methods have been explored to realize ultrasensitive, specific, and user-friendly biosensors. Furthermore, NAzyme-based bioassays have already broken into the in vitro diagnostics market, with more promise in the pipeline.
Subject(s)
Biosensing Techniques , DNA, Catalytic , Biosensing Techniques/trends , DNA, Catalytic/metabolism , RNA/metabolism , RNA, Catalytic/metabolismABSTRACT
When screening microbial populations or consortia for interesting cells, their selective retrieval for further study can be of great interest. To this end, traditional fluorescence activated cell sorting (FACS) and optical tweezers (OT) enabled methods have typically been used. However, the former, although allowing cell sorting, fails to track dynamic cell behavior, while the latter has been limited to complex channel-based microfluidic platforms. In this study, digital microfluidics (DMF) was integrated with OT for selective trapping, relocation, and further proliferation of single bacterial cells, while offering continuous imaging of cells to evaluate dynamic cell behavior. To enable this, magnetic beads coated with Salmonella Typhimurium-targeting antibodies were seeded in the microwell array of the DMF platform, and used to capture single cells of a fluorescent S. Typhimurium population. Next, OT were used to select a bead with a bacterium of interest, based on its fluorescent expression, and to relocate this bead to a different microwell on the same or different array. Using an agar patch affixed on top, the relocated bacterium was subsequently allowed to proliferate. Our OT-integrated DMF platform thus successfully enabled selective trapping, retrieval, relocation, and proliferation of bacteria of interest at single-cell level, thereby enabling their downstream analysis.
ABSTRACT
In disease diagnostics, single- and multiplex nucleic acid (NA) detection, with the potential to discriminate mutated strands, is of paramount importance. Current techniques that rely on target amplification or protein-enzyme based signal amplification are highly relevant, yet still plagued by diverse drawbacks including erroneous target amplification, and the limited stability of protein enzymes. As a solution, we present a multicomponent nucleic acid enzymes (MNAzymes)-based system for singleplex and multiplex detection of NA targets in microwells down to femtomolar (fM) concentrations, without the need for any target amplification or protein enzymes, while operating at room temperature and with single base-pair resolution. After successful validation of the MNAzymes in solution, their performance was further verified on beads in bulk and in femtoliter-sized microwells. The latter is not only a highly simplified system compared to previous microwell-based bioassays but, with the detection limit of 180 fM, it is to-date the most sensitive NAzyme-mediated, bead-based approach, that does not rely on target amplification or any additional signal amplification strategies. Furthermore, we demonstrated, for the first time, multiplexed target detection in microwells, both from buffer and nasopharyngeal swab samples, and presented superior single base-pair resolution of this assay. Because of the design flexibility of MNAzymes and direct demonstration in swab samples, this system holds great promise for multiplexed detection in other clinically relevant matrices without the need for any additional NA or protein components. Moreover, these findings open up the potential for the development of next-generation, protein-free diagnostic tools, including digital assays with single-molecule resolution.
Subject(s)
Biosensing Techniques/methods , DNA/analysis , Nucleic Acid Amplification Techniques/methods , Base Pairing , Humans , Limit of Detection , Magnets/chemistry , Nasopharynx/chemistrySubject(s)
Acculturation , Individuality , Students , Adult , Female , Humans , Male , Students/psychology , UniversitiesABSTRACT
DNA- and MNAzymes are nucleic acid-based enzymes (NAzymes), which infiltrated the otherwise protein-rich field of enzymology three decades ago. The 10-23 core NAzymes are one of the most widely used and well-characterized NAzymes, but often require elevated working temperatures or additional complex modifications for implementation at standard room temperatures. Here, we present a generally applicable method, based on thermodynamic principles governing hybridization, to re-engineer the existing 10-23 core NAzymes for use at 23 °C. To establish this, we first assessed the activity of conventional NAzymes in the presence of cleavable and non-cleavable substrate at 23 °C as well as over a temperature gradient. These tests pointed towards a non-catalytic mechanism of signal generation at 23 °C, suggesting that conventional NAzymes are not suited for use at this temperature. Following this, several novel NAzyme-substrate complexes were re-engineered from the conventional ones and screened for their performance at 23 °C. The complex with substrate and substrate-binding arms of the NAzymes shortened by four nucleotides on each terminus demonstrated efficient catalytic activity at 23 °C. This has been further validated over a dilution of enzymes or enzyme components, revealing their superior performance at 23 °C compared to the conventional 10-23 core NAzymes at their standard operating temperature of 55 °C. Finally, the proposed approach was applied to successfully re-engineer three other new MNAzymes for activity at 23 °C. As such, these re-engineered NAzymes present a remarkable addition to the field by further widening the diverse repertoire of NAzyme applications.
Subject(s)
DNA, Catalytic/chemistry , DNA, Catalytic/metabolism , DNA, Single-Stranded/chemistry , DNA, Single-Stranded/metabolism , DNA/chemistry , Protein Engineering , Catalysis , Reproducibility of Results , Substrate Specificity , Temperature , ThermodynamicsABSTRACT
OBJECTIVES: Acculturation is a multidimensional process involving changes in behaviour and beliefs. Questionnaires developed to measure acculturation are typically designed for specific ethnic populations and adult experiences. This study developed a questionnaire that measures acculturation among ethnically diverse populations of youth that can be included as a module in population surveys. METHODS: Questionnaires measuring acculturation in youth were identified in the literature. The importance of items from the existing questionnaires was determined using a Delphi process and this informed the development of our questionnaire. The questionnaire was then pilot tested using a sample of 248 Canadians aged 18-25 via an online system. Participants identified as East and South East Asian (27.8%), South Asian (17.7%) and Black (13.7%). The majority were 1st (33.5%) or 2nd generation immigrants (52.0%). After redundant items were eliminated, exploratory factor analysis grouped items into domains, and, for each domain, internal consistency, and convergent validity with immigrant generation then age at immigration estimated. A subset of participants re-completed the questionnaire for reliability estimation. RESULTS: The literature review yielded 117 articles that used 13 questionnaires with a total of 440 questions. The Delphi process reduced these to 32 questions. Pilot testing occurred in 248 Canadians aged 18-25. Following item reduction, 16 questions in three domains remained: dominant culture, heritage language, and heritage culture. All had good internal consistency (Cronbach's alphas > .75). The mean dominant domain score increased with immigrant generation (1st generation: 3.69 (95% CI: 3.49-3.89), 2nd: 4.13 (4.00-4.26), 3rd: 4.40 (4.19-4.61)), and mean heritage language score was higher among those who immigrated after age 12 than before (p = .0001), indicative of convergent validity. CONCLUSIONS: This Bicultural Youth Acculturation Questionnaire has demonstrated validity. It can be incorporated into population health surveys to elucidate the impact of acculturation on health outcomes among bicultural youth.