Memes are a popular online communication tool that are participatory, playful and contextual in nature. While the use of meme creation as an education tool in higher education has been limited, meme creation requires students to reflect on material presented in the classroom, synthesize new content from learned concepts, and present it in a contextualized manner. The objective of this study was to examine the level of reflection demonstrated in a meme creation assignment introduced into a master's level nutrition course which covered systemic and ethical issues related to nutrition, research, and clinical practice. Participants were graduate-level dietetic students (n = 55) enrolled in an Evidence Based Practice course in a Master of Science/Dietetic Internship Program. Students were instructed to create a meme about content from the course. Memes were analyzed for depth of reflection using an existing quantitative framework and researchers conducted a hybrid thematic analysis. Of the 82 memes submitted, 9 (11%) were rated as reflection level 0 (description), 11 (13.4%) as level 1 (reflective description), 26 (31.7%) as level 2 (dialogic reflection), 15 (18.3%) as level 3 (transformative reflection) and 21 (25.6%) as level 4 (critical reflection). Four primary themes identified were Ethics, Philosophy of Science, Art of Science and Science and the Public. Our findings confirm that the use of a meme creation assignment is an educational tool that promotes student reflection. Students displayed a higher-than-expected level of transformative or critical reflection which may be due to the playful, visual format of memes.
Memetic algorithms achieve strong optimization performance by combining population-based global search with local refinement operators, yet their effectiveness critically depends on the design and management of memes. Local search strategies are typically handcrafted, problem-specific, and fixed prior to execution. This paper proposes a fourth-generation memetic framework in which Large Language Models (LLMs) are embedded directly into the optimization loop as adaptive generators of local search operators. At each triggering point, the LLM receives a structured state vector encoding the current search dynamics and generates a candidate meme in the form of an executable Python function. Generated operators are subject to a two-stage validation procedure combining semantic similarity assessment and implementation-level comparison, ensuring that only sufficiently novel and syntactically correct operators are admitted to a dynamically growing meme library. A cooldown-regulated triggering mechanism balances periodic and stagnation-based generation, while a probability-weighted selection policy prioritizes newly generated memes without discarding previously validated ones. The proposed framework is evaluated on the CEC 2017 benchmark suite for continuous black-box optimization and compared against classical memetic algorithms. Experimental results demonstrate that the LLM-driven approach consistently outperforms non-LLM memetic baselines, confirming the viability of generative language models as adaptive heuristic components within population-based optimization.
Generation Z has different requirements, problems, expectations, and attitudes that they bring into the educational environment. Teachers have unprecedented challenges in addressing their needs and aspirations. Gamification involves integrating game-like elements into an existing educational framework. Rather than being a standalone teaching method, it serves as an instructional design approach. In this research, we investigated the impact of a meme-based approach in teaching various neuroanatomy topics and compared its effectiveness with traditional methods. In addition, we explored whether gender differences or students' academic levels influence their learning outcomes. The present research was conducted with third-year medical students enrolled in the neuroscience module at the College of Medicine, AlMaarefa University, Riyadh, Saudi Arabia, from September to December 2023, during the academic year 2023-2024. Students were instructed to create memes to aid in the comprehension and retention of anatomical facts and structures throughout the course. After each exam, we divide the questions into meme-related and non-meme-related categories. We then use the exam item analysis to compare the total student responses to the questions in the two categories, assess their performance, and compare the results between the male and female sections. Both male and female students showed significant improvement in their performance when responding to questions about meme-based material. Among high-achieving students, males showed a significant improvement in performance; however, the improvement among girls did not reach statistical significance. Conversely, both male and female students in the lower-performing group exhibited considerable improvements when interacting with meme-augmented topics. In conclusion, a meme-based approach significantly improved students' grades compared to the traditional approach; its effect was more pronounced among lower achievers and males. Although our findings suggest that male students may gain a greater advantage from this approach, the evidence for a gender-based moderating effect remains preliminary. The observed patterns require verification through larger, multicenter studies.
Ageism, characterized by stereotypes, prejudices, and discrimination based on age, remains a pervasive societal issue with significant social and psychological implications. This study explores the use of memes-a popular and relatable digital medium-as a creative and reflective tool for understanding how adolescents conceptualize aging and ageism following an educational workshop. The intervention involved 727 middle school students (ages 11-15) who were introduced to the concept of ageism and then created memes to express their understanding. Visual and thematic analyses of 274 student-generated memes revealed diverse perceptions of older persons: 168 memes (74%) challenged ageist stereotypes by incorporating more positive views of aging, while 59 memes (26%) reinforced existing ageist perceptions. The study offers methodological contributions, demonstrating the potential of meme creation as a way to capture nuances in students' perceptions of older age and ageism.
Internet memes increasingly move from social media into physical product packaging, yet little is known about how consumers respond when online cultural symbols become package design cues. Drawing on the Stimulus-Organism-Response framework, this study examines how meme-based packaging shapes purchase intention and sharing intention through perceived value, brand warmth, and cultural resonance. A between-subjects survey experiment was conducted with 305 Chinese adult consumers, who evaluated either a meme-based packaging stimulus or a no-explicit-meme conventional packaging control stimulus. Partial least squares structural equation modeling showed that purchase intention and sharing intention followed different dominant mechanisms. Perceived value was the strongest predictor of purchase intention, whereas cultural resonance was the strongest predictor of sharing intention. Visual attractiveness most strongly enhanced perceived value, while playfulness and expression-product fit contributed more clearly to brand warmth and cultural resonance. Mediation results further showed that brand warmth and cultural resonance consistently transmitted the effects of meme-packaging cues, whereas the value route was more selective. These findings show how online cultural symbols can continue to shape consumer evaluation and social transmission after entering physical product interfaces.
This article examines how racialised memes circulating on X revive scientific racism, demonstrating that hybrid formats reconfigure biological essentialisation within contemporary digital culture. Using a corpus of 68 viral items and 4 case studies, our analysis applies Shifman's content-form-stance model, informed by cognitive psychology. We show that these memes adopt common scientific conventions while relying on visual polarisation, whereby rapid perceptual contrasts stand in for arguments. Familiar templates, reinforced by selective algorithmic amplification, create a sense of perceptual legitimacy through fast categorisation and pre-attentive processing. We also trace how these cognitive shortcuts provide the basis for a regime of racialisation tailored to digitally immersed male audiences, producing gendered behavioural stereotypes and disappearance anxieties. A further pattern is the recoding of older racial hierarchies into neutral-sounding language of competence, allowing these ideas to travel across racialist, masculinist, techno-elitist, and anti-DEI (Diversity, Equity, Inclusion) communities. In this environment, memes work as micro-infrastructures of classification that align claims to objectivity with affect and platform incentives, making inequality appear natural and necessary. We conclude that effective critique must meet these images on their own terrain, with a pedagogy of the gaze that teaches publics how to recognise and challenge persuasion in meme-driven spaces.
Eight children with normal neurological examinations in a child neurology clinic were observed to have an abnormal posturing of the arms, head and torso after being instructed to run down a clinic hallway. The posturing was involuntary, unconscious, stereotyped, subject to voluntary control, and had the appearance identical to that of the anime character Naruto. None of the children knew each other, and each watched Naruto on television. Their abnormal posturing was not evidence of an organic disorder but instead of a cultural meme with which, in the era of widely disseminated social media, the clinician should be aware in order to avoid unnecessary evaluation and misdiagnosis.
The study explored community perspectives on ethical considerations in clinical research, focusing on the ESRIFAL trial conducted in Cameroon. The hermeneutic phenomenological approach was used to analyze and interpret the data. Participants were selected by systematic sampling. In-depth interviews were used to gather data from the participants. Some findings revealed that while ethical principles such as informed consent are highly recommended in clinical trials, participants did not understand and interpret these documents with as much importance as expected. Participants had other priorities and objectives for engaging in the trial; getting rid of the disease onchocerciasis was one of them. Signing the forms was nothing more than a formality for them. In conclusion, the study emphasizes the importance of strengthening ethical oversight mechanisms, but more importantly, the fact that ethical principles need to be contextualized to fit into the social and cultural realities of the societies in which trials are implemented.
Seatbelt-related breast trauma is an uncommon consequence of motor vehicle collisions. Most injuries present as localized contusion or hematoma, whereas progressive breast hematomas requiring surgical intervention are exceedingly rare, particularly in patients without coagulation abnormalities. Obesity may further alter trauma biomechanics and influence hematoma progression. A 49-yearold woman with obesity (Body Mass Index: 38 kg/m2) presented to the emergency department one hour after a motor vehicle collision with left breast swelling, firmness, and ecchymosis. Body temperature, blood pressure, and heart rate were within normal limits. Contrast-enhanced computed tomography (CT) demonstrated a 62×52 mm breast hematoma without evidence of arterial extravasation. Conservative treatment with compression therapy was continued for three days; however, pain and breast tension progressively increased, and ultrasonography demonstrated mild enlargement of the hematoma. On the fourth day, minimally invasive surgical evacuation was performed, yielding a large volume of organized hematoma. Postoperatively, symptoms improved rapidly. At two-week follow-up, breast symmetry was fully restored with no residual deformity. This case represents a rare presentation of seatbelt-related breast trauma: a progressive breast hematoma without active bleeding that ultimately required surgical evacuation. Obesity may have contributed to the failure of conservative management and earlier clinical progression. Early recognition of symptom worsening is essential to optimize cosmetic and functional outcomes. Emniyet kemerine bağlı meme travması, motorlu araç kazaları sonrasında nadir görülen bir tablodur. Olguların çoğu lokalize kontüzyon veya hematom şeklinde ortaya çıkar ve genellikle konservatif yöntemlerle izlenebilir. Bununla birlikte, progresif seyreden ve cerrahi müdahale gerektiren meme hematomları oldukça nadirdir; özellikle koagülasyon bozukluğu bulunmayan hastalarda bu durum daha da nadir bildirilmektedir. Obezite, darbe biyomekaniğini değiştirebilir; bu nedenle hematomun genişlemesi, semptomların progresyonu ve konservatif tedaviye yanıt üzerinde etkili olabilir. VKİ 38 kg/m2 olan 49 yaşındaki obez kadın hasta, motorlu araç kazasından 1 saat sonra sol memede şişlik, sertlik ve morluk ile acil servise başvurdu. Vücut ısısı, kan basıncı ve kalp hızı normaldi. Kontrastlı BT’de arteriyel ekstravazasyon saptanmaksızın 62×52 mm meme hematomu izlendi. Kompresyon tedavisi 3 gün uygulandı; ancak ağrı ve meme gerginliği arttı ve ultrasonografide hematomda büyüme görüldü. Dördüncü günde minimal insizyonla cerrahi olarak hematom boşaltıldı. Postoperatif dönemde semptomlar hızla geriledi. İki haftalık takipte meme simetrisi rezidü deformite olmaksızın tamamen düzeldi. Bu olgu, aktif kanama bulgusu olmaksızın progresyon gösteren ve nihayetinde cerrahi gerektiren emniyet kemerine bağlı meme travmasının nadir bir prezentasyonudur. Obezite, konservatif tedavinin başarısızlığına ve daha erken klinik progresyona yatkınlık oluşturabilir. Bulgularda kötüleşmenin erken fark edilmesi, optimal kozmetik ve fonksiyonel sonuçların elde edilmesi için kritik öneme sahiptir.
Tongue-tie and upper lip-tie have been linked to breastfeeding difficulties; however, traditional evaluation tools rely on subjective assessments. Ultrasound imaging enables objective assessment of infant sucking dynamics. This study aimed to assess the impact of tongue-tie and upper lip-tie on sucking function in newborns, using ultrasound imaging and other evaluation tools. This comparative observational study with short-term follow-up included 67 newborns (25 with tongue-tie, 21 with upper lip-tie, and 21 controls) and their mothers. Oral examinations were conducted during the hospital stay, prior to discharge. The maxillary labial frenulum was evaluated using the Kotlow classification, and the lingual frenulum was assessed with the Tongue Tie and Breastfed Infants (TABBY) tool. Sucking function was examined using ultrasound imaging between postnatal Days 5 and 15. Breastfeeding was evaluated via the LATCH tool, and maternal nipple pain was rated using a numeric scale. A 1-month follow-up call gathered data on feeding status, nipple pain, and surgical interventions. No significant differences in LATCH or nipple pain scores were found across groups. Ultrasound measurements in the tongue-up position showed significantly lower anterior tongue height in the tongue-tie group. In the same position, intraoral space depth was reduced significantly in the upper lip-tie group. Anterior tongue height correlated negatively with maternal nipple pain in the tongue-tie group (r = -0.49, p = 0.014). Tongue-tie and upper lip-tie do not always lead to observable breastfeeding problems. Ultrasound imaging offers valuable insights into subtle sucking dysfunctions not captured by clinical assessment. Arka plan:Dil bağı ve üst dudak bağı emzirme zorluklarıyla ilişkilendirilmiştir; ancak geleneksel değerlendirme araçları öznel değerlendirmelere dayanmaktadır. Ultrason görüntüleme, bebeklerin emme dinamiklerinin objektif olarak değerlendirilmesini mümkün kılar.Araştırmanın Amacı:Bu çalışma, yenidoğanlarda dil bağı ve üst dudak bağının emme fonksiyonu üzerindeki etkisini ultrason görüntüleme ve diğer değerlendirme araçlarını kullanarak incelemeyi amaçladı.Yöntemler:Bu kısa dönem takipli karşılaştırmalı gözlemsel çalışmaya 67 yenidoğan (25 dil bağı, 21 üst dudak bağı ve 21 kontrol) ve anneleri dahil edildi. Ağız muayeneleri, hastanede kalış sırasında taburculuk öncesinde gerçekleştirildi. Maksiller labial frenulum Kotlow sınıflandırmasına göre, lingual frenulum ise Tongue Tie and Breastfed Infants (TABBY) aracı ile değerlendirildi. Emme fonksiyonu postnatal 5. ve 15. günler arasında ultrason görüntüleme ile incelendi. Emzirme, LATCH aracı ile değerlendirildi ve annedeki meme ağrısı sayısal derecelendirme ölçeği ile puanlandı. Doğumdan bir ay sonra yapılan takip telefon görüşmesinde beslenme durumu, meme ağrısı ve cerrahi müdahalelerle ilgili veriler toplandı.Bulgular:Gruplar arasında LATCH veya meme ağrısı puanlarında anlamlı bir fark bulunmadı. Dil yukarı pozisyonunda yapılan ultrason ölçümlerine göre, dil bağı grubunda ön dil yüksekliği diğer gruplara göre anlamlı derecede düşüktü. Aynı pozisyonda, üst dudak bağı grubunda intraoral boşluk derinliği hem dil bağı grubuna hem de kontrol grubuna kıyasla azalmıştı. Dil bağı grubunda, dil yukarı pozisyonunda iken, ön dil yüksekliği ile meme ağrı skoru arasında anlamlı ve negatif bir ilişki gözlendi (r = -0,49; p = 0,014).Sonuç:Dil bağı ve üst dudak bağı her zaman gözlemlenebilir emzirme sorunlarına neden olmaz. Ultrason görüntüleme, klinik değerlendirmede belirlenmeyen emme fonksiyonu bozuklukları hakkında değerli bilgiler sağlar.
Previous research has provided valuable insights into how virtual likes and comments on self-presentational content affect users' emotions and self-evaluations, yet less is known about the causal attributions people make in response to such feedback. This study analyzed responses to an open-ended question to explore the types of causal attributions individuals make for favorable and unfavorable feedback on different types of Instagram posts - selfies, elsies (self-images taken by others), and memes. Undergraduate participants (N = 125; 77% female; 81% 17- to 24-years-old) were randomly assigned to one of three image conditions (selfie, elsie, or meme) and told their selected image would be posted to a university-affiliated Instagram account. After a 5-minute distraction period, during which participants believed their image was posted for feedback, they were told they had received either favorable or unfavorable (fabricated) feedback. They were asked to write down the perceived causes of the feedback they received in an open-ended question. Additionally, participants reported their perception of societal attitudes toward selfies compared to elsies, to explore whether these perceptions may have influenced their attributions. Findings revealed that self-focused images (selfies and elsies) more frequently prompted internal attributions - especially appearance-related - compared to meme posts, which elicited primarily external attributions tied to audience reception. Despite recognizing society's negative perceptions of selfie-posters, participants rarely attributed feedback to these broader social norms. The results suggest that posting self-images can heighten self-awareness and self-objectification, leading individuals to internalize social feedback. These attribution patterns carry implications for self-esteem and self-presentation. Understanding how users interpret feedback may inform efforts to promote healthier engagement with social media.
Amyotrophic Lateral Sclerosis (ALS) is a progressive neurodegenerative disease characterized by motor neuron degeneration, oxidative stress, neuroinflammation, and neurotransmitter imbalances. This study explored the neuroprotective potential of melatonin (MLT), alone and in combination with edaravone (EDR), in a methylmercury (MEME)-induced ALS rat model. MEME exposure effectively replicated ALS pathology, causing behavioral deficits, oxidative stress, neuroinflammation, apoptosis, and widespread structural damage in critical brain regions and the spinal cord. MLT administration at 5 mg/kg (MLT5) and 10 mg/kg (MLT10) significantly mitigated MEME-induced neurotoxicity in a dose-dependent manner. MLT improved motor function, reduced depressive-like behavior, and restored body weight. Biochemically, MLT enhanced antioxidant defenses, including superoxide dismutase (SOD) and catalase (CAT), reduced pro-inflammatory cytokines, interleukin-1 beta (IL-1β), increased anti-inflammatory cytokines, interleukin-10 (IL-10), and restored neurotransmitter balance like dopamine and Gamma-Aminobutyric Acid (GABA). Mechanistically, MLT activated the IGF-1 signaling pathway, promoting neuronal survival and reducing apoptosis (Caspase-3 expression). Histopathological analyses confirmed that MLT preserved neuronal and glial integrity, reduced demyelination, and restored myelin basic protein (MBP) levels in brain and cerebrospinal fluid. The combination of MLT and EDR exhibited synergistic neuroprotective effects, surpassing the efficacy of individual treatments in reducing oxidative stress, inflammation, and neuronal damage. Behavioral and biochemical improvements were paralleled by systemic recovery, as evidenced by normalized hematological parameters and reduced methylmercury accumulation in brain tissues. These findings underscore MLT, particularly in combination with EDR, as a potent therapeutic agent for ALS, offering multi-targeted neuroprotection. Future studies should explore its translational potential in clinical settings for the treatment of neurodegenerative diseases.
Chikungunya virus (CHIKV) is a re-emerging alphavirus that has caused millions of cases worldwide, yet its molecular epidemiology in Saudi Arabia remains poorly understood. This study integrates bioinformatic analysis of the envelope gene (E1) gene sequences from Saudi isolates with global genotypes to characterize circulating lineages, selection pressures and stability effects of endemic mutations. A total of 109 CHIKV E1 sequences (1155 bp) representing the East/Central/South African (ECSA), Asian, and West African genotypes were retrieved from GenBank and GISAID. Phylogenetic relationships were reconstructed using maximum likelihood (IQ-TREE). Codon-based selection analyses were performed with MEME, FEL, SLAC, and FUBAR. The structural effects of nine missense mutations were assessed using DynaMut and consensus predictors (DUET, mCSM). All seven Saudi isolates clustered within the ECSA-Indian Ocean Lineage (IOL) subclade with strong bootstrap support (≥95%). Short branch lengths among Saudi strains indicated recent common ancestry and limited local divergence, suggesting possible repeated introductions or limited local circulation. No codon showed robust evidence of positive selection across multiple methods. However, episodic diversifying selection was detected at codon 99 (MEME, p = 0.01), while pervasive purifying selection acted on numerous sites (e.g., codons 135, 307, 344; strong signals across FEL, SLAC, and FUBAR). A single conserved N-linked glycosylation site was present at residue 141 (NITV motif) in all Saudi and most global strains. Three mutations unique to or prominent in Saudi isolates (N20H, L136F, A249T) were identified; consensus stability predictions (DUET) classified them as destabilizing. Saudi CHIKV strains belong exclusively to the ECSA-IOL lineage and exhibit strong purifying selection on the E1 gene, consistent with functional constraints on this essential fusion protein. The identified Saudi-associated mutations appear to be non-adaptive, tolerated changes. These findings underscore the value of continued genomic surveillance to monitor potential adaptive evolution, particularly in the context of mass gatherings and competent Aedes vectors.
This study proposes a quantitative design transformation framework to revitalize Shanxi's historical opera stages by aligning heritage semantics with consumer needs. A multi-criteria decision-making model integrating architectural meme theory, AHP, QFD, TOPSIS, and Stable Diffusion was employed. Cultural 'genes' extracted from 3D scans formed a meme lineage, where AHP and QFD quantified attributes to guide generative design, and TOPSIS facilitated performance evaluation. Of the resulting cultural products, the "Spire-Top Penholder" yielded the highest relative affinity coefficient, successfully balancing structural authenticity with modern utility. The study confirms that reinterpreting elements like flying eaves through product semantics augments the public perception of heritage value. This study proposes a data-driven paradigm for sustainable heritage preservation, providing a scalable framework for integrating historical architecture into digital platforms for cultural documentation, interpretation, and dissemination.
Chikungunya virus (CHIKV) is re-emerging globally, yet integrative analyses combining E1-based phylodynamics, full-genome recombination screening, and genomic characterization of Chinese isolates remain limited. We curated 1,291 high-quality CHIKV full genomes from GenBank (through September 2025). Alignments were generated with MAFFT and models selected using ModelFinder. Time-scaled phylogenetic reconstruction and Bayesian skyline analyses were conducted in BEAST using the E1 coding region. Recombination was screened across complete genomes using RDP4 and further evaluated using SimPlot. Codon-based selection analyses of the Chinese structural coding-region subset were performed using GARD, BUSTED, MEME, and SLAC. For 84 Chinese isolates, variation in the structural protein region was quantified, and amino-acid substitutions were mapped onto AlphaFold-predicted protein structures. Root-to-tip regression of the E1 dataset showed apparent temporal structure, but this signal was interpreted cautiously because of lineage structure and uneven outbreak-driven sampling. Three of sixteen recombination candidates were retained as sequence-supported signals after additional evaluation, with retained breakpoints localized mainly to nsP2 and E2. Among available recent genomes, the ECSA lineage was prominently represented, including genomes associated with recent outbreaks in China. Structural proteins were overall conserved, with amino-acid variability concentrated mainly in E2 and E1. The E1-based analyses provided a cautious time-calibrated framework for CHIKV evolutionary history. Recombination signals were rare after validation, with retained breakpoints localized mainly to nsP2 and E2, and the ECSA lineage was prominently represented among recent available genomes. In Chinese isolates, structural proteins were overall conserved, with amino-acid variability concentrated in E2 and E1. Codon-based analyses supported predominant purifying selection with limited evidence of episodic selection at a subset of sites.
Vertebrate evolution from a single multipurpose cloaca to separate urogenital and anal openings remains a central molecular question with relevance to reproductive diversification and anorectal malformations. We investigated BMP7, a critical regulator of cloacal formation, across 37 vertebrates from cartilaginous fish to mammals using phylogenetics, sequence alignment, physicochemical assessment, domain analysis, motif analysis and molecular docking. The BMP7 core TGF-beta domain was highly conserved (>90% identity), whereas peripheral regions showed significant habitat-associated divergence (p < 0.01), including shifts in isoelectric point between aquatic and terrestrial vertebrates (pI 6.22-6.97 to 6.98-8.98) and altered stability indices. A key finding was that placental mammals, in which cloacal septation produces distinct anorectal and urogenital outlets, showed lineage-specific amino-acid changes, including Pro111Ala, linked to predicted BMP7-FOXA1 regulatory refinement. MEME analysis identified mammal-bird GWQDWI conservation and amphibian-reptile RACGCH divergence, supporting peripheral BMP7 adaptation during cloacal evolution.
The Small Auxin Up-Regulated RNA (SAUR) gene family has been implicated in various physiological processes in plants, including auxin signaling, senescence, and stress tolerance. Our previous genome-wide transcriptional analyses on postharvest senescence in broccoli (Brassica oleracea L.) identified the BoSAUR gene family as a key regulator in this process. In this study, we examined the expression patterns of BoSAUR family genes under various treatments aimed at delaying postharvest senescence of broccoli, analyzing its protein structure and phylogenetic relationship within the BoSAUR family. We applied gene expression, protein structure, phylogenetic analysis, and mutant characterization to investigate SAUR genes and their roles in senescence. Genome sequences of broccoli were retrieved for identifying BoSAUR genes, followed by phylogenetic, motif, and gene structure analyses using tools like MEME and GSDS. Promoter regions were analyzed with PlantCARE, and chromosomal locations were mapped using TBtools. A T-DNA insertion mutant of SAUR21 gene in Arabidopsis was analyzed for functional analysis. We found that BoSAUR21-like (LOC106327409) was consistently downregulated during natural and postharvest senescence across all treatments, regardless of ethylene status, identifying it as a core senescence-associated gene in broccoli. Genome-wide analysis identified that the BoSAUR21-like gene family shares a single-exon architecture with conserved SAUR motifs. To infer the function of BoSAUR21-like, we characterized the Arabidopsis AtSAUR21 T-DNA insertion mutant, a homologue of BoSAUR21-like. Phenotypic analysis of this mutant revealed accelerated leaf senescence, early bolting, and enhanced chlorophyll degradation compared to wild-type, suggesting that the orthologous BoSAUR21-like gene family likely plays a conserved role in senescence regulation in broccoli. These findings establish BoSAUR21-like gene as a promising candidate for genetic manipulation aimed at delaying postharvest senescence in broccoli and other Brassica crops. Future work should include direct functional characterization of BoSAUR21-like in broccoli through CRISPR-Cas9 knockout and overexpression approaches to confirm its role and assess its potential for extending postharvest shelf life.
Oropouche virus (OROV) is an emerging virus that causes a wide spectrum of febrile illnesses in humans. In this study, we examined two of the most commonly used primer sets for the clinical diagnosis of OROV to identify potential reduction in diagnostic sensitivity or even escape due to genetic variability in the target regions of circulating strains. We also evaluated selective pressures on the S segment to determine the optimal region for selecting diagnostic targets. For this, a total of 225 OROV S segment sequences (2015-2024) from multiple American countries were retrieved from NCBI and aligned with commonly used RT-PCR primers to identify mismatches. Subsequently, sequences (1960-2024) were analyzed for evolutionary studies, using HyPhy (Datamonkey), applying aBSREL, MEME, and FUBAR to assess branch- and site-level selection. Alignment of 225 OROV S segment sequences revealed widespread mismatches in primer binding regions. Selection pressure analysis showed no branch-level positive selection, one site under episodic diversifying selection, and 81 sites under purifying selection. Notably, highly conserved regions did not overlap with commonly targeted primer sites. These findings suggest that current diagnostic assays may be vulnerable to reduced sensitivity due to genetic variability. Continuous genomic surveillance and comprehensive primer design studies are essential to ensure reliable detection of emerging OROV strains.
Ranunculus L., the largest genus of Ranunculaceae, exhibits remarkable ecological diversity, yet genomic resources for this genus remain limited. Here we report the complete chloroplast (cp) genomes of Ranunculus arvensis L. and Ranunculus laetus Wall. ex Hook.f. & Thomson. The R. laetus assembly is the first cp genome reported for this species, whereas the R. arvensis assembly, generated from a Pakistani population, provides an independent accession that complements the recently released GenBank record PV621859 from China. Both genomes were analyzed comparatively against 22 previously published Ranunculus cp genomes. The cp genomes ranged from 154,474 to 158,314 bp in length and displayed a typical quadripartite structure. All analyzed species harbored 112 unique genes, comprising 78 protein-coding genes, 30 transfer RNA genes, and four ribosomal RNA genes. The cp genomes were highly conserved in terms of gene order, guanine-cytosine content, codon usage, amino acid composition, and simple sequence repeats. Phylogeny-aware selection analysis models (BUSTED, MEME, FUBAR) revealed that, despite overall purifying selection at the gene level, codon-based FUBAR detected pervasive positive selection across ~34.6% of plastid genes; matK and ycf1 emerged as the most prominent hotspots for both episodic and pervasive adaptation. Nucleotide diversity analyses revealed several highly variable regions, notably rpl32-ndhF, ycf1, and ndhF. Phylogenetic analyses based on complete cp genome sequences and complementary single-gene trees resolved two major clades corresponding to biogeographic patterns: Clade I comprised predominantly Eurasian and East Asian taxa, whereas Clade II comprised a transcontinental assemblage of East Asian, North American, and Himalayan species. The R. laetus and R. arvensis were grouped in Clade II. Overall, these results elucidate the conserved genomic architecture, evolutionary dynamics, and phylogenetic relationships of Ranunculus cp genomes and provide valuable genomic resources for future studies on phylogeny, taxonomy, DNA barcoding, and population genetics within Ranunculaceae.
Chronic myeloid leukemia (CML) is a hematological malignancy that has its cause as Philadelphia chromosome and BCR: ABL fusion gene. Although specific drugs such as imatinib have been used to a great effect to enhance the outcome of treatment, resistance is still a challenge especially in later stages of the disease. This work has explored the phenotypic role of eight single nucleotide variants (SNVs) in the downstream sequence of MIR5195 gene, which are determined in real-life sample of patients. The initial experiment involved whole-exome sequencing (WES) to identify genetic variants in advanced phase CML patients and Sanger sequencing to validate the variants. The results indicate the possible importance of the established mutations as well as mutations that are yet to be characterized in the regulation of MIR5195 and their potential impact on the development of CML. Comprehensive bioinformatics analyses (MEME, TOMTOM, and RegulomeDB) revealed that these mutations profoundly reconfigure local cis-regulatory architecture, resulting in the loss of conserved transcription factor binding motifs and the emergence of a mutation-specific motif enriched for hematopoietic and cancer-associated transcription factors, including RXRB, FOXP1, GATA-1, and GATA-2. Three-dimensional structural modeling using AlphaFold3 showed that both wild-type and mutant downstream MIR5195 sequences retain canonical B-form DNA, but the mutant exhibits localized changes in backbone curvature and groove geometry within the mutation-enriched motif. Molecular docking analyses revealed transcription factor–specific redistribution of binding affinities rather than uniform loss of interaction, with increased binding observed for GATA1 (− 242.20 vs. − 228.26) and RXRB (− 223.57 vs. − 221.44), and reduced binding for FOXP1 (− 213.63 vs. − 219.93) and GATA2 (− 242.15 vs. − 281.57). These findings indicate selective modulation of transcription factor engagement driven by downstream non-coding mutations. Based on the emergence of a mutation-specific regulatory motif, we designed a transcription factor decoy oligonucleotide targeting the altered binding site as a potential mutation-directed regulatory strategy. Overall, this integrative computational and experimental framework demonstrates that downstream MIR5195 mutations quantitatively reprogram transcription factor binding networks in CML and highlights non-coding regulatory variation as a biologically meaningful contributor to leukemia pathogenesis.