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29 pages, 1637 KB  
Article
Rethinking Performance Evaluation: Strategic Alignment in the Service Sector Through a Case-Based Framework
by Maria C. Tavares and Mariana Vaz
Adm. Sci. 2025, 15(10), 390; https://doi.org/10.3390/admsci15100390 - 8 Oct 2025
Abstract
Performance management is critical for aligning human capital with organizational strategy, particularly in the increasingly competitive service sector. However, universally effective performance appraisal systems (PASs) exist, as effectiveness depends on contextual and organizational specificities. In Portugal, where services account for nearly three-quarters of [...] Read more.
Performance management is critical for aligning human capital with organizational strategy, particularly in the increasingly competitive service sector. However, universally effective performance appraisal systems (PASs) exist, as effectiveness depends on contextual and organizational specificities. In Portugal, where services account for nearly three-quarters of gross value added, PAS implementation remains underdeveloped, highlighting a gap between strategic intent and practice. This study aims to address that gap by investigating how a performance appraisal model can be tailored to the service sector. A case study was conducted at PCI—Creative Science Park, S.A., a consulting firm, using a qualitative approach. The research design combined a literature review to identify theoretical dimensions of performance evaluation with an employee questionnaire to capture organizational perceptions and priorities. Integration of both strands of evidence informed the construction of the framework. The findings indicate that employees value objective-based evaluation as the most relevant dimension, complemented by customer feedback, adaptive performance, and organizational citizenship. Furthermore, the integration of 360° feedback mechanisms and regular review cycles emerged as key enablers of fairness and engagement. By combining theoretical insights with employee perspectives, this study contributes to a customized and flexible PAS that enhances strategic alignment in the service sector. The proposed model provides both scholarly value, by advancing the discussion on context-specific PAS design, and practical value, by offering a reference for organizations seeking to align human performance with mission-critical outcomes. Full article
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13 pages, 1564 KB  
Article
Pan-Resistant HIV-1 Drug Resistance Among Highly Treated Patients with Virological Failure on Dolutegravir-Based Antiretroviral Therapy in Zimbabwe
by Tendai Washaya, Benjamin Chimukangara, Justin Mayini, Sandra Bote, Nyasha Chin’ombe, Shungu Munyati and Justen Manasa
Viruses 2025, 17(10), 1348; https://doi.org/10.3390/v17101348 - 8 Oct 2025
Abstract
The HIV-1 epidemic continues to challenge global public health, especially in sub-Saharan Africa. The rise in drug-resistant viruses, particularly pan-resistant strains, threatens treatment effectiveness, hindering progress toward UNAIDS viral suppression goals. This is critical in low-to-middle income countries (LMICs) like Zimbabwe, where treatment [...] Read more.
The HIV-1 epidemic continues to challenge global public health, especially in sub-Saharan Africa. The rise in drug-resistant viruses, particularly pan-resistant strains, threatens treatment effectiveness, hindering progress toward UNAIDS viral suppression goals. This is critical in low-to-middle income countries (LMICs) like Zimbabwe, where treatment options and access to drug resistance testing are limited. This cross-sectional study analyzed 102 genotypes from patients with HIV-1 RNA ≥ 1000 copies/mL after at least 6 months on a dolutegravir (DTG)-based ART. HIV-1 genotyping and drug resistance interpretation were performed using the Stanford HIV Drug Resistance Database. Overall, 62% of genotypes harbored at least one drug resistance mutation, with 27% showing integrase strand transfer inhibitor (INSTI)-associated mutations. High-level resistance to DTG and cabotegravir was found in 14% and 23% of integrase sequences, respectively, primarily driven by G118R and E138K/T mutations. Pan-resistance was observed in 18% of complete genotypes, with one case of four class resistance. These results highlight the emergence of INSTI resistance in LMICs. The study underscores the urgent need for enhanced HIV drug resistance testing, continuous surveillance, and strategic optimization of ART regimens in resource-constrained settings to ensure effective HIV management. Full article
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31 pages, 10340 KB  
Article
Silencing the cyp314a1 and cyp315a1 Genes in the Aedes albopictus 20E Synthetic Pathway for Mosquito Control and Assessing Algal Blooms Induced by Recombinant RNAi Microalgae
by Xiaodong Deng, Changhao He, Chunmei Xue, Dianlong Xu, Juncai Li and Xiaowen Fei
Insects 2025, 16(10), 1033; https://doi.org/10.3390/insects16101033 - 7 Oct 2025
Abstract
As one of the key vectors for the transmission of Dengue fever, Aedes albopictus is highly ecologically adaptable. The development of environmentally compatible biological defence and control technologies has therefore become an urgent need for vector biological control worldwide. This study constructed and [...] Read more.
As one of the key vectors for the transmission of Dengue fever, Aedes albopictus is highly ecologically adaptable. The development of environmentally compatible biological defence and control technologies has therefore become an urgent need for vector biological control worldwide. This study constructed and used double-stranded RNA (dsRNA) expression vectors targeting the cyp314a1 and cyp315a1 genes of Ae. albopictus to transform Chlamydomonas reinhardtii and Chlorella vulgaris, achieving RNA interference (RNAi)-mediated gene silencing. The efficacy of the RNAi recombinant algal strain biocide against Ae. albopictus was evaluated by administering it to Ae. albopictus larvae. The results showed that the oral administration of the cyp314a1 and cyp315a1 RNAi recombinant C. reinhardtii/C. vulgaris strains was lethal to Ae. albopictus larvae and severely affected their pupation and emergence. The recombinant algal strains triggered a burst of ROS (Reactive Oxygen Species) in the mosquitoes’ bodies, resulting in significant increases in the activities of the superoxide dismutase (SOD), peroxiredoxin (POD) and catalase (CAT), as well as significant upregulation of the mRNA levels of the CME pathway genes in larvae. In the simulated field experiment, the number of Ae. albopictus was reduced from 1000 to 0 in 16 weeks by the RNAi recombinant Chlorella, which effectively controlled the population of mosquitoes. Meanwhile, the levels of nitrogen (N), phosphorus (P), nitrate, nitrite, ammonia and COD (Chemical Oxygen Demand) in the test water decreased significantly. High-throughput sequencing analyses of 18S rDNA and 16S rDNA showed that, with the release of RNAi recombinant Chlorella into the test water, the biotic community restructuring dominated by resource competition caused by algal bloom, as well as the proliferation of anaerobic bacteria and the decline of aerobic bacteria triggered by anaerobic conditions, are the main trends in the changes in the test water. This study is an important addition to the use of RNAi recombinant microalgae as a biocide. Full article
(This article belongs to the Special Issue RNAi in Insect Physiology)
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16 pages, 909 KB  
Article
First Survey on Sea Turtles’ Interactions in Mussel Farms in Italy
by Ludovica Di Renzo, Giulia Mariani, Marco Matiddi, Cecilia Silvestri, Stefania Chiesa, Tommaso Petochi, Giovanna Marino, Federica Pizzurro, Simone Fazio, Emanuela Rossi, Giuseppe Prioli, Ike Olivotto and Giorgia Gioacchini
Animals 2025, 15(19), 2909; https://doi.org/10.3390/ani15192909 - 6 Oct 2025
Abstract
Sea turtles, particularly the opportunistic feeder species loggerhead turtles (Caretta caretta), are increasingly reported as a source of disturbance to mussel farming operations, especially in the Adriatic Sea. Despite the evident damage caused by these interactions, comprehensive national data on the [...] Read more.
Sea turtles, particularly the opportunistic feeder species loggerhead turtles (Caretta caretta), are increasingly reported as a source of disturbance to mussel farming operations, especially in the Adriatic Sea. Despite the evident damage caused by these interactions, comprehensive national data on the phenomenon are still lacking. This study aimed to address this gap through a survey conducted among Italian mussel farmers, combined with the analysis of gastrointestinal contents from stranded sea turtles along the Adriatic and Tyrrhenian coasts, focusing on the ingestion of Mediterranean mussels (Mytilus galloprovincialis). Survey results revealed frequent turtle sightings in the northern Adriatic (Veneto and Emilia-Romagna) during summer months (June to August), while southern regions (Molise and Puglia) reported more sightings in autumn (September to October), likely influenced by seasonal water temperatures. The Mediterranean mussel was identified as the most commonly ingested mollusk in the Adriatic, with a notable increase in presence from 2018 to 2021. Although mussels are not a targeted prey, they appear to be a consistent dietary component due to adaptive feeding behavior. These interactions are increasingly and consistently reported, leading to significant management challenges for mussel farms. Based on these findings, a broader national and international assessment is recommended to evaluate the overall impact of sea turtles on shellfish aquaculture in the Mediterranean. Full article
(This article belongs to the Section Aquatic Animals)
32 pages, 508 KB  
Article
The Reflections of Raa Haqi Cosmology in Dersim Folk Tales
by Ahmet Kerim Gültekin
Religions 2025, 16(10), 1274; https://doi.org/10.3390/rel16101274 - 6 Oct 2025
Viewed by 68
Abstract
This article illuminates the cosmology of Raa Haqi (often called Dersim Alevism or Kurdish Alevism), a rarely examined strand within Alevi Studies. Existing scholarship’s emphasis on identity politics and sparse ethnography has left Raa Haqi’s mythological and cosmological dimensions underexplored. This paper approaches [...] Read more.
This article illuminates the cosmology of Raa Haqi (often called Dersim Alevism or Kurdish Alevism), a rarely examined strand within Alevi Studies. Existing scholarship’s emphasis on identity politics and sparse ethnography has left Raa Haqi’s mythological and cosmological dimensions underexplored. This paper approaches Raa Haqi through a dual authority framework: (1) Ocak lineages and Ocak–talip relations—sustained by kinship institutions like kirvelik, musahiplik, and communal rites such as the cem—and (2) jiares, non-human agents from the Batın realm that manifest in Zahir as sacred places, objects, and animals. Methodologically, I conduct a close, motif-based reading of folktales compiled by Caner Canerik (2019, Dersim Masalları I), treating them as ethnographic windows into living theology. The analysis shows that tales encode core principles—rızalık (mutual consent), ikrar (vow), sır (the secret knowledge), fasting and calendrical rites, ritual kinship, and moral economies involving humans, animals, and Batın beings. Dreams, metamorphosis, and jiare-centered orientations structure time–space, ethics, and authority beyond the Ocak, including in individual re-sacralizations of objects and sites. I conclude that these narratives do not merely reflect belief; they actively transmit, test, and renew Raa Haqi’s cosmological order, offering Alevi Studies a theory-grounded, source-proximate account of Kurdish Alevi mythic thought. Full article
33 pages, 3027 KB  
Article
Bacillus subtilis DinG 3′⟶5′ Exo(ribo)nuclease: A Helpmate to Mitigate Replication Stress
by Begoña Carrasco, Rubén Torres, María López-Sanz, Rogelio Hernández-Tamayo, Peter L. Graumann and Juan C. Alonso
Int. J. Mol. Sci. 2025, 26(19), 9681; https://doi.org/10.3390/ijms26199681 - 4 Oct 2025
Viewed by 225
Abstract
Bacillus subtilis DinG/XPD-like paralogues, DinG and YpvA, have been implicated in overcoming replication stress. DinG possesses a DEDD exonuclease and DNA helicase domains, whereas YpvA lacks the DEDD exonuclease domain. We report that DinG·Mg2+ (hereafter referred to as DinG) degrades linear single-stranded [...] Read more.
Bacillus subtilis DinG/XPD-like paralogues, DinG and YpvA, have been implicated in overcoming replication stress. DinG possesses a DEDD exonuclease and DNA helicase domains, whereas YpvA lacks the DEDD exonuclease domain. We report that DinG·Mg2+ (hereafter referred to as DinG) degrades linear single-stranded (lss) DNA with 3′→5′ polarity and binds lssDNA with higher affinity than its exonuclease-deficient mutant DinG D10A E12A. DinG’s ssDNA-dependent ATPase activity neither stimulates nor inhibits DNA degradation. When bound to the 3′-end of forked DNA, DinG destabilises and degrades the substrate; however, in the presence of ATP, DinG dissociates before reaching the duplex junction. DinG degrades the RNA strand within RNA–DNA hybrids but does not cleave lssRNA unless complexed with Mn2+. DinG removes genomic R-loops, as RnhC and PcrA do. DinG physically interacts with RecA and PolA and functions in the same pathway as translesion synthesis (TLS) DNA polymerases (DNAPs) to respond to both spontaneous and methyl methanesulphonate (MMS)-induced mutagenesis. DinG-mGold forms spontaneous foci at or near replication forks, which become enriched following MMS or rifampicin treatment. We propose that DinG contributes to mitigating replication stress by degrading R-loop barriers and facilitating TLS, potentially via RecA-linked mechanisms. Full article
(This article belongs to the Section Molecular Biology)
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17 pages, 672 KB  
Review
Saying “Yes” to NONO: A Therapeutic Target for Neuroblastoma and Beyond
by Sofya S. Pogodaeva, Olga O. Miletina, Nadezhda V. Antipova, Alexander A. Shtil and Oleg A. Kuchur
Cancers 2025, 17(19), 3228; https://doi.org/10.3390/cancers17193228 - 3 Oct 2025
Viewed by 172
Abstract
Pediatric tumors such as neuroblastoma are characterized by a genome-wide ‘transcriptional burden’, surmising the involvement of multiple alterations of gene expression. Search for master regulators of transcription whose inactivation is lethal for tumor cells identified the non-POU domain-containing octamer-binding protein (NONO), a member [...] Read more.
Pediatric tumors such as neuroblastoma are characterized by a genome-wide ‘transcriptional burden’, surmising the involvement of multiple alterations of gene expression. Search for master regulators of transcription whose inactivation is lethal for tumor cells identified the non-POU domain-containing octamer-binding protein (NONO), a member of the Drosophila Behavior/Human Splicing family known for the ability to form complexes with macromolecules. NONO emerges as an essential mechanism in normal neurogenesis as well as in tumor biology. In particular, NONO interactions with RNAs, largely with long non-coding MYCN transcripts, have been attributed to the aggressiveness of neuroblastoma. Broadening its significance beyond MYCN regulation, NONO guards a subset of transcription factors that comprise a core regulatory circuit, a self-sustained loop that maintains transcription. As a component of protein–protein complexes, NONO has been implicated in the control of cell cycle progression, double-strand DNA repair, and, generally, in cell survival. Altogether, the pro-oncogenic roles of NONO justify the need for its inactivation as a therapeutic strategy. However, considering NONO as a therapeutic target, its druggability is a challenge. Recent advances in the inactivation of NONO and downstream signaling with small molecular weight compounds make promising the development of pharmacological antagonists of NONO pathway(s) for neuroblastoma treatment. Full article
(This article belongs to the Special Issue Precision Medicine and Targeted Therapies in Neuroblastoma)
29 pages, 2059 KB  
Review
Roles and Applications of Circular RNA in Virus Infection
by Fang Gou, Yanmei Gao, Keke Zhong, Tian Bu, Yinggang Li, Faxiang Li and Rong Yang
Int. J. Mol. Sci. 2025, 26(19), 9656; https://doi.org/10.3390/ijms26199656 - 3 Oct 2025
Viewed by 322
Abstract
Circular RNAs (circRNAs) are novel types of covalently closed single-stranded RNA formed by the backsplicing of precursor mRNAs (pre-mRNAs). Recently, circRNAs have been shown to play a crucial role in various diseases, including cancers, cardiovascular and cerebrovascular diseases, and autoimmune diseases. Accumulating evidence [...] Read more.
Circular RNAs (circRNAs) are novel types of covalently closed single-stranded RNA formed by the backsplicing of precursor mRNAs (pre-mRNAs). Recently, circRNAs have been shown to play a crucial role in various diseases, including cancers, cardiovascular and cerebrovascular diseases, and autoimmune diseases. Accumulating evidence has demonstrated that both host-derived and virus-encoded circRNAs play pivotal roles during viral infection, including modulating viral entry, genome replication, latency establishment, and the host antiviral immune responses while simultaneously facilitating viral immune evasion. However, their roles during viral infections and circRNA-host interactions remain to be further investigated. Therefore, this article reviews the key characteristics and biological functions of circRNAs, as well as recent advances in understanding the interactions between circRNAs from different sources and viral infections, which will offer insights for developing therapies targeting virus-associated diseases. Full article
(This article belongs to the Section Molecular Microbiology)
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14 pages, 4597 KB  
Article
Exogenous Application of IR-Specific dsRNA Inhibits Infection of Cucumber Green Mottle Mosaic Virus in Watermelon
by Yanhui Wang, Liming Liu, Yongqiang Fan, Yanli Han, Zhiling Liang, Yanfei Geng, Fengnan Liu, Qinsheng Gu, Baoshan Kang and Chaoxi Luo
Agronomy 2025, 15(10), 2332; https://doi.org/10.3390/agronomy15102332 - 2 Oct 2025
Viewed by 247
Abstract
Cucumber green mottle mosaic virus (CGMMV) represents a serious threat in the production of watermelon. Small RNAs facilitate a mechanism known as RNA interference (RNAi), which regulates gene expression. RNAi technology employs foreign double-stranded RNAs (dsRNAs) to target and reduce the expression levels [...] Read more.
Cucumber green mottle mosaic virus (CGMMV) represents a serious threat in the production of watermelon. Small RNAs facilitate a mechanism known as RNA interference (RNAi), which regulates gene expression. RNAi technology employs foreign double-stranded RNAs (dsRNAs) to target and reduce the expression levels of specific genes in plants by interfering with their mRNAs. In this study, watermelon plants were treated with dsRNAs of CGMMV MET, IR, and HEL fragments that had been generated in E. coli HT115. We investigated variations in several factors, including viral accumulation, virus-derived small interfering RNAs (vsiRNAs), and symptom severity. MET-dsRNA, IR-dsRNA and HEL-dsRNA dramatically decreased the symptoms of CGMMV in plants in the growth chamber test. Plants treated with viral-derived dsRNA showed a considerable decrease in both virus titers and vsiRNA levels. We also explored the mobility of spray-on dsRNA-derived long dsRNA and discovered that it could be identified in both inoculated leaves and the systemic leaves. IR-dsRNA outperformed MET-dsRNA and HEL-dsRNA in dsRNA therapy. Illumina sequencing of small RNAs from watermelon plants treated with IR-dsRNA and those that were not treated showed that the decreased accumulation of vsiRNAs was consistent with interference with CGMMV infection in systemic leaves. dsRNA-treated plants showed a higher level of 24-nt viral siRNA and lower level of 22-nt viral siRNA accumulation, while 22-nt viral siRNA predominated in untreated plants, indicating that dsRNA treatment improved DCL3 activity. In conclusion, our research provides deeper insights into the mechanism of antiviral RNA interference and confirms the effectiveness of applying dsRNA locally to enhance plant antiviral activity. Full article
(This article belongs to the Section Pest and Disease Management)
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21 pages, 2264 KB  
Article
Thermodynamic Determinants in Antibody-Free Nucleic Acid Lateral Flow Assays (AF-NALFA): Lessons from Molecular Detection of Listeria monocytogenes, Mycobacterium leprae and Leishmania amazonensis
by Leonardo Lopes-Luz, Paula Correa Neddermeyer, Gabryele Cardoso Sampaio, Luana Michele Alves, Matheus Bernardes Torres Fogaça, Djairo Pastor Saavedra, Mariane Martins de Araújo Stefani and Samira Bührer-Sékula
Biomolecules 2025, 15(10), 1404; https://doi.org/10.3390/biom15101404 - 2 Oct 2025
Viewed by 286
Abstract
Antibody-free nucleic acid lateral flow assays (AF-NALFA) are an established approach for rapid detection of amplified pathogens DNA but can yield inconsistent signals across targets. Since AF-NALFA depends on dual hybridization of probes to single-stranded amplicons (ssDNA), site-specific thermodynamic (Gibbs free energy-ΔG) at [...] Read more.
Antibody-free nucleic acid lateral flow assays (AF-NALFA) are an established approach for rapid detection of amplified pathogens DNA but can yield inconsistent signals across targets. Since AF-NALFA depends on dual hybridization of probes to single-stranded amplicons (ssDNA), site-specific thermodynamic (Gibbs free energy-ΔG) at probe-binding regions may be crucial for performance. This study investigated how site-specific-ΔG and sequence complementarity at probe-binding regions determine Test-line signal generation, comparing native and synthetic amplicons and assessing the effects of local secondary structures and mismatches. Asymmetric PCR-generated ssDNA amplicons of Listeria monocytogenes, Mycobacterium leprae, and Leishmania amazonensis were analyzed in silico and tested in AF-NALFA prototypes with gold-labeled thiol probes and biotinylated capture probes. T-line signals were photographed, quantified (ImageJ version 1.4k), and statistically correlated with site-specific-ΔG. While native ssDNA from M. leprae and L. amazonensis failed to produce AF-NALFA T-line signals, L. monocytogenes yielded strong detection. Site-specific-ΔG below −10 kcal/mol correlated with reduced hybridization. Synthetic oligos preserved signals despite structural constraints, whereas ~3–4 mismatches, especially at capture probe regions, markedly impaired T-line intensity. The performance of AF-NALFA depends on the synergism between thermodynamic accessibility, site-specific-ΔG-induced site constraints, and sequence complementarity. Because genomic context affects hybridization, target-specific thermodynamic in silico evaluation is necessary for reliable pathogen DNA detection. Full article
(This article belongs to the Section Molecular Biology)
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11 pages, 1301 KB  
Article
Monitoring Single DNA Docking Site Activity with Sequential Modes of an Optoplasmonic Whispering-Gallery Mode Biosensor
by Narima Eerqing, Ekaterina Zossimova, Sivaraman Subramanian, Hsin-Yu Wu and Frank Vollmer
Sensors 2025, 25(19), 6059; https://doi.org/10.3390/s25196059 - 2 Oct 2025
Viewed by 167
Abstract
In recent years, there has been rapid advancement in single-molecule techniques, driven by their unparalleled precision in studying molecules whose sizes are beyond the diffraction limit. Among these techniques, optoplasmonic whispering gallery mode sensing has demonstrated great potential in label-free single-molecule characterization. It [...] Read more.
In recent years, there has been rapid advancement in single-molecule techniques, driven by their unparalleled precision in studying molecules whose sizes are beyond the diffraction limit. Among these techniques, optoplasmonic whispering gallery mode sensing has demonstrated great potential in label-free single-molecule characterization. It combines the principles of localized surface plasmon resonance (LSPR) and whispering gallery mode (WGM) sensing, offering exceptional sensing capabilities, even at the level of single ions. However, current optoplasmonic WGM sensing operates in a multiplexed channel, making it challenging to focus on individual binding sites of analyte molecules. In this article, we characterize different binding sites of DNA analyte molecules hybridizing to docking strands on the optoplasmonic WGM sensor, using the ratio of the resonance shift between sequential polar WGM modes. We identify specific docking sites that undergo transient interactions and eventually hybridize with the complementary analyte strands permanently. Full article
(This article belongs to the Section Biosensors)
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11 pages, 2044 KB  
Communication
CRISPR-Enhanced Colorimetric Aptasensor for Adenosine Triphosphate Detection Based on MoS2-Based Nanozymes
by Zhiqiang Zhu, Haojie Ma, Huashan Yao, Yuan Yuan, Xiangyang Miao and Shao Su
Biosensors 2025, 15(10), 651; https://doi.org/10.3390/bios15100651 - 1 Oct 2025
Viewed by 221
Abstract
As the direct energy source in organisms, accurate and simple detection of adenosine triphosphate (ATP) is of great significance. Herein, a colorimetric aptasensor for ATP determination was designed by integrating the CRISPR/Cas12a system with an aptamer, and with Prussian blue nanocube and gold [...] Read more.
As the direct energy source in organisms, accurate and simple detection of adenosine triphosphate (ATP) is of great significance. Herein, a colorimetric aptasensor for ATP determination was designed by integrating the CRISPR/Cas12a system with an aptamer, and with Prussian blue nanocube and gold nanoparticle co-functionalized MoS2 (MoS2-PBNCs-AuNPs) nanozymes. As expected, the introduced CRISPR/Cas12a system and aptamer could efficiently amplify the detection signal and improve the specific recognition ability, respectively. Meanwhile, the catalytic activity of the MoS2-PBNCs-AuNPs nanozymes can be regulated with the concentration of ATP. The high-affinity binding of ATP to the aptamer competitively inhibited aptamer-crRNA hybridization, causing fewer Cas12 proteins to be activated. As a result, the uncleaved single-stranded DNA (ssDNA) adsorbed onto the surface of nanozymes to effectively enhance their catalytic oxidation capability toward 3,3′,5,5′-tetramethylbenzidine (TMB). According to this phenomenon, this CRISPR-enhanced colorimetric aptasensor can detect down to 0.14 μM ATP with high selectivity, reproducibility, and stability. In addition, acceptable recoveries and low relative standard deviations of the aptasensor for ATP determination suggest that it is promising for application in early detection of clinical-related diseases. Full article
(This article belongs to the Special Issue CRISPR/Cas System-Based Biosensors)
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18 pages, 2407 KB  
Article
Mitogenomic Characterization of Microhyla fissipes and Its Implications for Phylogenetic Analysis in Microhylidae
by Siqi Shan, Simin Chen, Chengmin Li, Huiling Huang, Yaqing Liao and Lichun Jiang
Biology 2025, 14(10), 1342; https://doi.org/10.3390/biology14101342 - 1 Oct 2025
Viewed by 240
Abstract
The microhylid frog Microhyla fissipes is a protected terrestrial wildlife species in China, recognized for its ecological, economic, and scientific value. However, its mitochondrial genome remains poorly characterized. To address this gap, we sequenced and annotated the complete mitogenome of M. fissipes to [...] Read more.
The microhylid frog Microhyla fissipes is a protected terrestrial wildlife species in China, recognized for its ecological, economic, and scientific value. However, its mitochondrial genome remains poorly characterized. To address this gap, we sequenced and annotated the complete mitogenome of M. fissipes to elucidate its structural organization and phylogenetic placement within Microhylidae. The assembled mitogenome is 16,723 bp in length and contains 37 genes, including 13 protein-coding genes, 2 rRNAs, and 22 tRNAs, along with one control region and the origin of heavy-strand replication. We also identified eight overlapping regions and eleven intergenic spacers. The overall base composition showed an A + T bias (59.91%) with negative AT-skew (−0.04) and GC-skew (−0.27). All tRNAs displayed typical cloverleaf secondary structures, except for trnS1, which lacked the D-arm. Phylogenetic reconstruction using both maximum likelihood and Bayesian inference strongly supported the monophyly of Microhylidae and revealed a sister-group relationship between Microhyla and Kaloula. Within Microhyla, M. fissipes was most closely related to M. heymonsi, with which it formed a well-supported clade that also included Microhyla okinavensis, Microhyla mixtura, and Microhyla beilunensis. Selection pressure analysis on protein-coding genes indicated widespread purifying selection (Ka/Ks < 1) across most genes, except for ATP8, COX2, and COX3, which may be under relaxed selective constraints. These findings offer valuable genomic resources for the conservation of M. fissipes and provide new insights into the phylogeny and evolution of microhylid frogs. Full article
(This article belongs to the Special Issue Progress in Wildlife Conservation, Management and Biological Research)
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21 pages, 3978 KB  
Article
Role of the Chaperone Protein 14-3-3η in Regulation of the Infection Dynamics of the Influenza A (H1N1) Virus
by Debarima Chatterjee, Partha Pratim Mondal, Anneshwa Bhattacharya and Alok Kumar Chakrabarti
Viruses 2025, 17(10), 1337; https://doi.org/10.3390/v17101337 - 30 Sep 2025
Viewed by 281
Abstract
The 14-3-3 protein family, which includes the isoforms η, γ, ε, θ, β, and ζ, is essential for controlling a number of pathways linked to DNA and RNA viruses, including HIV, influenza A virus (IAV), measles virus, HRSV, and double-stranded DNA viruses. TRIM32, [...] Read more.
The 14-3-3 protein family, which includes the isoforms η, γ, ε, θ, β, and ζ, is essential for controlling a number of pathways linked to DNA and RNA viruses, including HIV, influenza A virus (IAV), measles virus, HRSV, and double-stranded DNA viruses. TRIM32, an E3 ubiquitin ligase, has been reported to target IAV’s PB1 polymerase for species-specific degradation via ubiquitination. Notably, 14-3-3η binds to phosphorylated TRIM32, preventing its autoubiquitylation and forming soluble but inactive cytoplasmic aggregates that regulate TRIM32 levels. However, the functional link between 14-3-3η, TRIM32, and PB1 during viral infection remains unclear. In this study, we establish a mechanistic connection between 14-3-3η–TRIM32 and TRIM32–PB1 interactions in IAV (H1N1) infection. We demonstrate that 14-3-3η directly interacts with PB1, influencing viral replication. Using transient knockdown models, we show that 14-3-3η deficiency alters influenza virus-induced cytotoxicity, cell death, immune responses, and reactive oxygen species (ROS) production. Additionally, we observe a significant reduction in the soluble TRIM32 levels in 14-3-3η-deficient cells, which leads to increased PB1 accumulation and thus suggests a critical regulatory role for 14-3-3η in PB1 stability. Our findings reveal a novel function of 14-3-3η in influenza virus infection, demonstrating its role in PB1 regulation via TRIM32 and its impact on innate immune activation. This study highlights 14-3-3η as a possible target for antiviral treatments against influenza and offers fresh insights into the host–virus relationship. Full article
(This article belongs to the Special Issue Interplay Between Influenza Virus and Host Factors)
15 pages, 891 KB  
Article
Elabela Attenuates Doxorubicin-Induced Oxidative DNA Damage and Apoptosis in Rat Left Ventricular Myocardium
by Katarzyna Matusik, Katarzyna Kamińska, Izabela Janiuk, Kaja Kasarełło, Maja Owe-Larsson, Daniel Dylko and Agnieszka Cudnoch-Jędrzejewska
Biomedicines 2025, 13(10), 2407; https://doi.org/10.3390/biomedicines13102407 - 30 Sep 2025
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Abstract
Background: Doxorubicin, a widely used chemotherapeutic agent, has been shown to increase reactive oxygen species (ROS) levels, disrupting cellular homeostasis not only in cancer cells but also in healthy tissues, particularly in cardiomyocytes, which leads to chemotherapy-induced cardiotoxicity. Therefore, new strategies are continually [...] Read more.
Background: Doxorubicin, a widely used chemotherapeutic agent, has been shown to increase reactive oxygen species (ROS) levels, disrupting cellular homeostasis not only in cancer cells but also in healthy tissues, particularly in cardiomyocytes, which leads to chemotherapy-induced cardiotoxicity. Therefore, new strategies are continually being explored to mitigate these adverse effects. One such approach is the use of additional substances with cardioprotective properties during doxorubicin therapy. A promising candidate is elabela, a peptide of the apelinergic system, which may exert protective effects against doxorubicin-induced oxidative stress in cardiomyocytes. Objectives: This study aims to evaluate the modulatory effects of elabela on oxidative stress markers, malondialdehyde (MDA) and 8-hydroxy-2′-deoxyguanosine (8-OHdG) in the left ventricle of the myocardium following chronic doxorubicin administration in rats. Material and Methods: 32 male, 12-week-old Sprague-Dawley rats (SPRD) were randomly assigned to four experimental groups. For 28 days, all animals received continuous infusions (2.5 μL/h) via subcutaneously implanted osmotic pumps of 0.9% NaCl or elabela (40 μg/kg body weight/day or 200 μg/kg body weight/day). Simultaneously, animals were injected intraperitoneally 4 times at weekly intervals with 0.9% NaCl or DOX (3.5 mg/kg body weight). Next, the animals were sacrificed, and left ventricular (LV) cardiac tissue was collected for further analysis. MDA and 8-OHdG and elabela level in LV lysate were assessed by ELISA. The Ela expression in LV was quantified by Real-Time PCR. The TUNEL assay, labeled with a 5′-triphosphate strand, was used to assess the degree of apoptosis. Results: DOX treatment decreased both the Ela expression and elabela levels in the LV. Elabela administration at a dose of 200 µg/kg body weight/day significantly decreased ELA levels and Ela expression compared to the control group. The level of 8-OhdG was unexpectedly decreased in the DOX group compared to controls, while elabela treatment at both doses restored 8-OHdG levels observed in the control group. However, TUNEL staining demonstrated that elabela administration at 200 µg/kg body weight/day reduced the number of apoptotic cardiomyocytes compared to the DOX-only group, indicating a protective effect against DOX-induced apoptosis. The lower dose of 40 µg/kg body weight/day showed a moderate, non-significant attenuation of apoptosis. Conclusions: Elabela showed a protective effect against DOX-induced cardiomyocyte apoptosis in the LV by promoting processes that reduce oxidative stress in cardiac cells. Full article
(This article belongs to the Section Cell Biology and Pathology)
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