bluuebunny/crossref_metadata_2025 huggingface dataset:
Due esperienze didattiche di early-algebra con il DIST-M: dalla costruzione del problema all’implementazione in aula
Authors: Aceto Maria , De Santis Maria Lodina , Donatiello Angela , Lemmi Giuseppina , Manzoni Simona , Montervino Concetta Rosaria , Picariello Luca , Scaramuzzino Valeria , Vadalà Giovanna
Published: November 2024

La possibilità di approcciare al discorso algebrico mediante attività che mettano in luce diverse rappresentazioni permette agli studenti lo sviluppo del pensiero relazionale. Le esperienze presentate sono relative alla progettazione di attività a carattere fortemente immersivo che hanno coinvolto studenti di scuole secondarie di primo grado nello sviluppo della competenza argomentativa e del formalismo simbolico in ambito di teoria dei numeri. L’approccio narrativo e immersivo è stato possibile grazie all’uso del dispositivo didattico del DIST-M, mentre l’utilizzo di sistemi semiotici visuali ha favorito la formazione di un concetto in una prospettiva vygotskijana.
Filmlerde Süper Kahraman Kadınlar ve İdeoloji: Wonder Woman Örneği
Authors: Gültekin Gökhan
Published: November 2024

Bu araştırmanın amacı, Wonder Woman’ın görünüm kazandığı filmler üzerinden aktarılan ideolojileri (kapitalizm, feminizm, faşizm, ataerkillik vs.) açığa çıkarmaya ve anlamlandırmaya çalışmaktır. Bu amaçla nitel bir bakış açısıyla hareket eden araştırmanın evrenini süper kahramanların ana coğrafyası olan Amerika’daki filmlerde yer alan süper kahraman kadınlar oluşturmaktadır. Araştırmanın örneklemi ise bu evrenden ‘amaçlı’ olarak şeçilen Wonder Woman karakteridir. Örnekleme dâhil edilen Wonder Woman, ilk görünüm kazandığı 1941’den günümüze kadar geçen süreçte yer aldığı filmler üzerinden değerlendirilmiştir. Bu değerlendirmede Wonder Woman’ın yer aldığı animasyon ve çizgi filmler kapsam dışı bırakılmıştır. Bu çalışma, yer aldığı sinema veya televizyon yapımı filmler üzerinden Wonder Woman aracılığıyla aktarılan ideolojileri ortaya koymaktadır. Ayrıca tarihsel süreçteki toplumsal değişimlerin süper kahraman kadınlara nasıl yansıyabileceğini göstermeye çalışması açısından önemlidir. Çalışmada Wonder Woman aracılığıyla ortaya koyulan ideolojileri analiz etmek üzere film çözümleme yöntemlerinden ‘ideolojik eleştiri’ ve ‘tarihsel eleştiri’ bir arada kullanılmıştır. Sonuç olarak, Wonder Woman’ın feminizmin savunuculuğunu yapacak şekilde motive edilmediği ortaya çıkmaktadır. Wonder Woman ataerkilliği, kapitalizmi ve ırkçılığı yansıtmaktadır. Ayrıca verili gerçekliğin kabulüne hizmet ettiği ve kadına dayatılan toplumsal rollerin yeniden üretimine yönelik değerler ürettiği gözlemlenmektedir.
Integrating artificial intelligence in strabismus management: current research landscape and future directions
Authors: Wu Dawen , Huang Xi , Chen Liang , Hou Peixian , Liu Longqian , Yang Guoyuan
Published: November 2024

Advancements in artificial intelligence (AI) are transforming strabismus management through improved screening, diagnosis, and surgical planning. Deep learning has notably enhanced diagnostic accuracy and optimized surgical outcomes. Despite these advancements, challenges such as the underrepresentation of diverse strabismus types and reliance on single-source data remain prevalent. Emphasizing the need for inclusive AI systems, future research should focus on expanding AI capabilities with large model technologies, integrating multimodal data to bridge existing gaps, and developing integrated management platforms to better accommodate diverse patient demographics and clinical scenarios.
Oxidative damage in Echinochloa crus−galli seeds exposed to Diaporthe sp. (Diaporthales, Ascomycota) fungal extract during germination
Authors: Somala Naphat , Manichart Nutcha , Laosinwattana Chamroon , Wichittrakarn Pattharin , Yoneyama Kaori , Teerarak Montinee , Chotsaeng Nawasit
Published: November 2024

This study investigates the inhibitory effects of Diaporthe sp. isolate EC010 extract on barnyardgrass (Echinochloa crus-galli) seed germination and growth. Application of sequential extraction techniques to Diaporthe sp. mycelium resulted in partial separation of the phytotoxic compounds. The ethyl acetate (EtOAc) fraction most greatly reduced seed germination (81.01%), root length (89.18%), and shoot length (84.74%) compared to the control. Chemical characterization using gas chromatography-mass spectrometry revealed major constituents of linoleic acid, butyl ester (9.69%), hexadecanoic acid (7.99%), and 14-pentadecenoic acid (7.86%). With regard to physiological and biochemical indexes, treated seeds exhibited lower imbibition, significantly decreased α-amylase (EC 3.2.1.1) activity (p<0.05), and increased accumulation of malondialdehyde (85.52%) and hydrogen peroxide (141.10%). Moreover, activity of the antioxidant enzymes superoxide dismutase (EC 1.15.1.1) and guaiacol peroxidase (EC 1.11.1.7) was upregulated (67.24 and 61.62%, respectively), while catalase (EC 1.11.1.6) activity was downregulated (-33.75%). The inference is that an imbalance in ROS levels combined with reduced antioxidant potential drives the gradual accumulation of oxidative damage in seed cells and consequent loss of seed viability. All told, these results confirm the Diaporthe extract to induce oxidative stress and inhibit antioxidant enzymes. This study clearly demonstrates the oxidative damage associated with Diaporthe allelochemicals.
Impact of UPF2 on the levels of CD81 on extracellular vesicles
Authors: Oh Chaehwan , Mazan-Mamczarz Krystyna , Gorospe Myriam , Noh Ji Heon , Kim Kyoung Mi
Published: November 2024

Extracellular vesicles (EVs) are involved in cell-to-cell communication. Following uptake, EV cargo molecules, including DNA, RNA, lipids, and proteins, influence gene expression and molecular signaling in recipient cells. Although various studies have identified disease-specific EV molecules, further research into their biogenesis and secretion mechanisms is needed for clinical application. Here, we investigated the role of UPF2 in regulating the biogenesis and components of EVs. Notably, UPF2 promoted the expression of CD81, a membrane protein marker of EVs, as UPF2 silencing decreased CD81 levels in EVs, both inside the cell and secreted. In contrast, the expression levels of CD63 increased, without altering the size or numbers of EVs. In addition, reducing UPF2 levels did not affect the total number of EVs but lowered production of CD81-positive EVs and reduced the efficiency of uptake by recipient cells. Collectively, our findings uncover a novel function for UPF2 in regulating the production of CD81 and changing EV properties.
Anticariogenic activity of marine brown algae Padina boergesenii and its active components towards Streptococcus mutans
Authors: Sangavi Ravichellam , Malligarjunan Nambiraman , Satish Lakkakula , Raja Veerapandian , Pandian Shunmugiah Karutha , Gowrishankar Shanmugaraj
Published: November 2024

Streptococcus mutans is a well-recognized bacterium that plays a predominant role in the progression of dental caries. Its pathogenicity is linked to several key characteristics, including the ability to produce organic acids (acidogenicity), thrive in low pH environments (aciduricity), synthesize exopolysaccharides (EPS) via glucosyltransferases, and form retentive biofilms. The treatment of dental caries with conventional antibiotics is often ineffective due to the bacterium’s capacity to form recalcitrant biofilms. To address these challenges, strategies that specifically target the pathogen’s virulence without affecting its viability have emerged as promising alternatives. In this context, we investigated the anticariogenic properties of the methanolic extract of Padina boergesenii (MEPB). MEPB demonstrated substantial, dose-dependent antibiofilm activity, with a maximum inhibition of 93% at 128 μg/mL, without compromising the viability of S. mutans. Anti-virulence assays using sub-MIC (minimum inhibitory concentration) levels of MEPB showed significant reductions in key virulence factors: 75% reduction in sucrose-dependent adherence, 65% reduction in sucrose-independent adherence, along with notable decreases in acid production, acid tolerance, and water-insoluble (85%) and water-soluble (52%) glucan synthesis. Additionally, MEPB significantly reduced cell surface hydrophobicity (55%) and extracellular DNA (eDNA) production (64%). qPCR analysis corroborated these in vitro findings, revealing that MEPB suppresses the expression of genes involved in S. mutans virulence, particularly genes related to EPS synthesis (gtfB, gtfC & gtfD) biofilm formation(gbpB & gbpC) and two-component regulatory system (vicR) were downregulated. Toxicity testing on human buccal epithelial cells confirmed the non-toxic nature of MEPB, suggesting its safety for potential therapeutic use. Furthermore, GC-MS/MS analysis identified palmitic acid, myristic acid, and stearic acid as the major active constituents of the MEPB extract. Subsequent biofilm inhibitory assays confirmed the potent antibiofilm efficacy of these compounds: palmitic acid (85%), myristic acid (72%) and stearic acid (83%). In conclusion, this study identifies P. boergesenii and its active biomolecules as potential anticariogenic agents, offering an alternative approach to combat dental caries by targeting bacterial virulence mechanisms rather than viability.
Correlation between remnant cholesterol and premature coronary artery disease and the severity of coronary artery lesions in men: a retrospective study
Authors: Dong Xingming , Chen Ke , Li Xiuqin , Tang Yuanyuan , Zhang Rui , Wang Jian
Published: November 2024

Objective

To investigate the correlation between remnant cholesterol (RC) and premature coronary artery disease (PCAD) and the severity of coronary artery lesions in men.

Methods

A total of 630 male subjects who underwent coronary angiography were included in the study. The general data, laboratory tests, and coronary angiography data of each group were statistically analyzed, and RC levels were calculated. According to the Gensini score, PCAD was divided into mild and severe lesion groups. The relationship between RC and PCAD and coronary artery lesions was analyzed using multivariate logistic regression and spearman correlation analysis, and the predictive value of RC for coronary artery lesions was evaluated using receiver operating characteristic (ROC) curves.

Results

The RC levels in the PCAD group were significantly higher than those in the non-PCAD group (p < 0.05), and RC was an independent risk factor for PCAD (p < 0.05).The RC levels in the severe lesion group were higher than those in the mild lesion group (p < 0.05), and RC levels were positively correlated with the Gensini score (r = 0.335, p < 0.001).Multivariate logistic regression analysis showed that RC was an independent risk factor for severe coronary artery lesions (p < 0.05).The ROC curve calculated the value of RC in predicting severe coronary artery lesions, with an area under the curve of 0.693, a cutoff value of 0.485 mmol/L, a sensitivity of 64.7%, and a specificity of 66.2%.

Conclusion

RC is an independent risk factor for PCAD and the severity of coronary artery lesions in adult men. RC levels are positively correlated with the severity of coronary artery lesions and can be used as an auxiliary indicator for clinical assessment of PCAD.
Safety and efficacy of outpatient intravenous diuresis in decompensated heart failure: a systematic review
Authors: Kalkur Roshni S. , Hintz John P. , Pathangey Girish , Manning Katharine A.
Published: November 2024

Introduction

Heart failure (HF) burdens the US healthcare system, with annual costs exceeding $30 billion. Outpatient intravenous (OP IV) diuresis in clinic or home settings may potentially improve outcomes and reduce costs, though limited data exists. This systematic review evaluates the safety, efficacy, and outcomes of OP IV diuresis in managing decompensated HF as a hospitalization alternative.

Methods

Following PRISMA 2020 guidelines, this systematic review used MeSH terms in MEDLINE, SCOPUS, CINAHL Complete, and Cochrane Central. From 16 selected studies, 15 were single-center; 6 prospective, 9 retrospective; and 1 was a randomized trial comparing OP IV diuresis to oral home regimen. Demographics, visit data, and outcomes were collected, and 30-day outcomes were compared to inpatient IV (IP IV) diuresis from 2021 Medicare HF hospitalizations.

Results

The review included 1,590 unique patients treated with OP IV diuretics, with a mean age of 70 ± 6 years, 69.7% male, and 74.8% NYHA III-IV. Minimal adverse post-diuresis events such as hypokalemia, hypotension, and worsening renal function occurred (4.5%, 0.7%, and 2.3% respectively). Post-visit mean weight loss was −2.2 ± 1.1 kg. The 30-day readmission rate for OP IV diuresis was significantly lower than IP IV diuresis (20.0% vs. 22.6%; p = 0.0.401), and 30-day mortality was also lower (5.6% vs. 10.7%; p = 0.003).

Discussion

OP IV diuresis is a safe and effective treatment for decompensated HF with minimal risk of adverse events. Data demonstrate reduced 30-day readmission, mortality rates, cost. These findings highlight the potential of OP IV diuresis as an enhanced alternative HF care; however, further randomized control trials are needed to evaluate long-term outcomes.
Opportunities, challenges and future perspectives of using bioinformatics and artificial intelligence techniques on tropical disease identification using omics data
Authors: Vidanagamachchi S. M. , Waidyarathna K. M. G. T. R.
Published: November 2024

Tropical diseases can often be caused by viruses, bacteria, parasites, and fungi. They can be spread over vectors. Analysis of multiple omics data types can be utilized in providing comprehensive insights into biological system functions and disease progression. To this end, bioinformatics tools and diverse AI techniques are pivotal in identifying and understanding tropical diseases through the analysis of omics data. In this article, we provide a thorough review of opportunities, challenges, and future directions of utilizing Bioinformatics tools and AI-assisted models on tropical disease identification using various omics data types. We conducted the review from 2015 to 2024 considering reliable databases of peer-reviewed journals and conference articles. Several keywords were taken for the article searching and around 40 articles were reviewed. According to the review, we observed that utilization of omics data with Bioinformatics tools like BLAST, and Clustal Omega can make significant outcomes in tropical disease identification. Further, the integration of multiple omics data improves biomarker identification, and disease predictions including disease outbreak predictions. Moreover, AI-assisted models can improve the precision, cost-effectiveness, and efficiency of CRISPR-based gene editing, optimizing gRNA design, and supporting advanced genetic correction. Several AI-assisted models including XAI can be used to identify diseases and repurpose therapeutic targets and biomarkers efficiently. Furthermore, recent advancements including Transformer-based models such as BERT and GPT-4, have been mainly applied for sequence analysis and functional genomics. Finally, the most recent GeneViT model, utilizing Vision Transformers, and other AI techniques like Generative Adversarial Networks, Federated Learning, Transfer Learning, Reinforcement Learning, Automated ML and Attention Mechanism have shown significant performance in disease classification using omics data.
Effect of air-particle abrasion and methacryloyloxydecyl phosphate primer on fracture load of thin zirconia crowns: an in vitro study
Authors: Alshali Ruwaida Z. , Alqahtani Mohammed A. , Alturki Basem N. , Algizani Loay I. , Batarfi Abdullah O. , Alshamrani Zuhair K. , Faden Razan M. , Bukhary Dalea M. , Altassan Mosa M.
Published: November 2024

The study aimed to investigate the effects of airborne-particle abrasion (APA) and 10-methacryloxydecyl dihydrogen phosphate (10-MDP primer) surface treatments on the fracture load of thin zirconia crowns made from 3Y-TZP and 5Y-PSZ zirconia. Eighty full-contour zirconia crowns of 0.5 mm thickness were fabricated from 3Y-TZP and 5Y-PSZ zirconia. Crowns of each material were divided into four groups based on the surface treatment applied to the fitting surface (n = 10): Group 1 (control), Group 2 (10-MDP Primer Only), Group 3 (APA Only), and Group 4 (10-MDP Primer + APA). Crowns were cemented using self-adhesive resin cement and subjected to thermocycling. Fracture load tests were performed using a universal testing machine with a hemispherical indenter. Statistical analysis was performed using one-way ANOVA, Tukey's post hoc test, and independent samples T-test (α = 0.05). The fracture load of 3Y-TZP was significantly higher than 5Y-PSZ crowns across all groups (P ≤ 0.001). Group 1 had the lowest fracture load, while Group 4 had the highest for both materials. In 3Y-TZP, the fracture load of Group 2 increased by 40% (P = 0.002) and Group 3 by 50% (P < 0.001) compared to Group 1. Group 4 showed a 90% increase over Group 1 (P < 0.001). For 5Y-PSZ, fracture load of Group 4 increased by 70% compared to Group 1 (P < 0.001). It was concluded that applying a 10-MDP primer or APA significantly increases the fracture load of thin 3Y-TZP zirconia crowns, with the combination of both treatments yielding the highest values. For 5Y-PSZ, a significant increase in fracture load is observed only when both APA and the 10-MDP primer are used together.
The effect of seaweed fertilisation on sulfur isotope ratios (δ34S) and grain size in barley: implications for agronomy and archaeological research
Authors: Blanz Magdalena , Gröcke Darren R. , Martin Peter , Church Mike J.
Published: November 2024

Introduction

Stable sulfur isotope ratios (δ34S) in bone collagen are often employed to study the consumption of marine and freshwater fish, wetland grazing, marine foraging patterns, and the possible geographic origins of food sources. However, a recent small-scale crop experiment showed that biofertilisation with seaweed can elevate δ34S in Celtic beans by ca. 10 ‰. Consumption of this food could erroneously suggest a marine diet and therefore has important implications for the reconstruction of past diets and dietary origins. However, limited research has so far been undertaken on cereals.

Methods

To address this issue, a large-scale field trial was undertaken on the Orkney Islands, whereby bere barley (a Scottish landrace, Hordeum vulgare L.) was biofertilised with seaweed at different dosages (25 t/ha, 50 t/ha), with a mineral NPK fertiliser, and left unfertilised as a control.

Results

The total barley biomass yield was higher and barley grains were enlarged following all fertilisation treatments compared to the control barley. Barley grain and straw from seaweed-fertilised crops had more elevated δ34S values by around 2–3 ‰ compared to unfertilised plants, while the NPK-fertilised grains and plants had δ34S values 1 ‰ lower.

Discussion

These results confirm previous hypotheses that seaweed fertilisation can elevate cereal δ34S values. The comparatively small δ34S difference between control and seaweed fertilised crops in this field trial is likely due to background elevated δ34S values in the soil (+12.7 ‰), which in turn may be due to long-term exposure to oceanic-influenced rain and sea spray and/or possible historical application of seaweed, or the underlying bedrock composition. The results of this study show that seaweed fertilisation can increase barley grain sizes and δ34S values, and thus should be considered when reconstructing land management and dietary practices in the archaeological record.
Why not ask them? A systematic scoping review of research on dyadic teacher–student relationships as perceived by students with emotional and behavioral problems
Authors: Vösgen-Nordloh Meike , Kulawiak Pawel R. , Bolz Tijs , Koomen Helma M. Y. , Hennemann Thomas , Leidig Tatjana
Published: November 2024

Introduction

Meta-analytical findings indicate that high-quality dyadic teacher-student relationships (TSRs) can act as social protective factors against the development or persistence of emotional and behavioral problems (EBPs) by fostering students’ social-emotional and cognitive learning. However, previous research primarily focused on samples of students without EBPs and relied on teacher-rated TSRs. Research on dyadic TSRs from the perspective of students with EBPs is scarce, yet their self-perceptions could offer valuable insights into whether and how dyadic TSRs serve as protective factors for them. Therefore, this systematic scoping review is guided by the central question of how research on dyadic TSRs from the perspective of students with EBPs has been conducted and what insights have been gained to date.

Methods

This question is explored through a narrative synthesis of existing studies, thereby identifying current research approaches, empirical findings, practical implications, and future research needs. A database search using keywords related to EBPs and TSRs yielded 24 included studies.

Results and discussion

Synthesizing these studies reveals five overarching conclusions. First, students with EBPs and their teachers tend to experience dyadic TSRs less favorably than typically developed (TD) students and their teachers. Second, relationship-enhancing interventions focusing on either teacher-student interactions, contextual factors or individual characteristics of teachers and students appear to be effective for students with EBPs. Third, students with EBPs seem to perceive stronger positive affective relationships with their teachers than teachers do with them. Fourth, students with EBPs may perceive their dyadic TSRs as ambivalent, exhibiting both highly positive and highly negative aspects. Fifth, for the social–emotional and academic development of students with EBPs, dyadic TSR-quality seems to be a risk or protective factor, acting differently than in TD-student. However, due to the diverse nature of the included studies, these conclusions remain only preliminary. Consequently, the review concludes with 10 key recommendations that might guide future research on dyadic TSRs from the perspective of students with EBPs.
The impact of current-loop control parameters on the electromagnetic transient voltage performance of voltage-source converter
Authors: Ding Xiaoyu , Chen Hongwei , Zhou Xiaobo , Shi Qianhong , Wang Guanzhong
Published: November 2024

Since the large-scale integration of renewable energy sources into the AC grid has led to a relative decline in the voltage support capacity of the grid and the deterioration of the voltage dynamic at the grid connection point, especially under fast-scale conditions, the voltage disturbance has become more obvious. To improve the dynamic characteristics of the electromagnetic transient voltage at the grid connection point, this paper uses a practical dynamic damping method to analyze the impact of the converter current inner loop, feedforward voltage, and other links on the dynamic performance of the electromagnetic transient voltage. First, the current inner loop dynamic in the converter’s synchronous coordinate is converted into an equivalent transfer function in the stationary coordinate, and the transfer function between the transient voltage disturbance at the grid connection point and the inner loop current output is established. On this basis, the Bode diagram and the vector diagram of the transfer function in the weak damping frequency band are used to analyze the dynamic damping of the current inner loop parameters and voltage feedforward filter parameters on the voltage disturbance at the grid connection point. The results indicate that moderately increasing the current inner loop bandwidth or reducing the feedforward filter bandwidth can help enhance the electromagnetic transient voltage stability of the grid connection point, but increasing the current inner loop bandwidth will worsen the low-frequency damping characteristics and reduce the feedforward filter bandwidth will still help increase low-frequency damping.
IL7R, GZMA and CD8A serve as potential molecular biomarkers for sepsis based on bioinformatics analysis
Authors: Li Jin , Wang Lantao , Yu Bin , Su Jie , Dong Shimin
Published: November 2024

Purpose

Sepsis is an unusual systemic reaction to what is sometimes an otherwise ordinary infection, and it probably represents a pattern of response by the immune system to injury. However, the relationship between biomarkers and sepsis remains unclear. This study aimed to find potential molecular biomarkers, which could do some help to patients with sepsis.

Methods

The sepsis dataset GSE28750, GSE57065 was downloaded from the GEO database, and ten patients with or without sepsis from our hospital were admitted for RNA-seq and the differentially expressed genes (DEGs) were screened. The Metascape database was used for functional enrichment analysis and was used to found the differential gene list. Protein-protein interaction network was used and further analyzed by using Cytoscape and STRING. Logistic regression and Correlation analysis were used to find the potential molecular biomarkers.

Results

Taking the intersection of the three datasets yielded 287 differential genes. The enrichment results included Neutrophil degranulation, leukocyte activation, immune effectors process, positive regulation of immune response, regulation of leukocyte activation. The top 10 key genes of PPI connectivity were screened using cytoHubba plugin, which were KLRK1, KLRB1, IL7R, GZMA, CD27, PRF1, CD8A, CD2, IL2RB, and GZMB. All of the hub genes are higher expressed in health group of different databases. Logistic regression showed that IL7R, GZMA and CD8A proteins were analyzed and all of them were statistically significant. Correlation analysis showed that there was a statistically significant correlation between IL7R, GZMA and CD8A.

Conclusion

KLRK1, KLRB1, IL7R, GZMA, CD27, PRF1, CD8A, CD2, IL2RB, GZMB are key genes in sepsis, which associated with the development of sepsis. However, IL7R, GZMA and CD8A may serve as the attractively potential molecular biomarkers for sepsis.
Polyfunctional T cells and unique cytokine clusters imprint the anti rAAV2/rAAV9 vector immune response
Authors: Holtkamp Stephan J. , Lagoda Florian R. , Lister Adam , Harish Pradeep , Kleymann Ulrike , Pesch Theresa , Soon Chai Fen , Pirmohamed Munir , Naisbitt Dean , Trautwein Mark
Published: November 2024

Polyfunctional T cells programmed to perform activities such as degranulation of lytic enzymes and simultaneous production of multiple cytokines are associated with more effective control of viral infections. Immune responses to recombinant adeno-associated virus (rAAV) vector delivery systems can critically influence therapeutic efficacy and safety of gene therapy. However, knowledge of polyfunctional T cells in anti-AAV immune responses is scarce. To bridge this knowledge gap, we have investigated the polyfunctionality of primary human CD4 T cells from healthy donors after in-vitro exposure to rAAV2 or rAAV9 vectors. By performing proliferation assays of co-cultured T cells and rAAV pulsed monocyte-derived dendritic cells from healthy donors we demonstrate T cell reactivity of 43% and 50% to rAAV2 and rAAV9 vectors, respectively. We validated this frequency in a second screen using another set of healthy donors measuring CD25 and CD71 T cell activation. Single T cell secretome analysis of reactive donors uncovered a Th1 pro-inflammatory, cytolytic and chemoattractive cytokine release profile after stimulation with rAAV2 or rAAV9 vectors. 12.4% and 9.6% of the stimulated T cells displayed a polyfunctional cytokine response, respectively, including elevated polyfunctional inflammatory indices. These responses were characterized by cytokine clusters such as Granzyme B, MIP1-α and TNF-α released in combination by single T cells. Overall, our results provide insights into adaptive immunity with rAAV vector serotypes which will be important in advancing gene therapy safety, vector selection, immunogenicity assessment and better patient selection for AAV gene therapy.
Characteristics of cytokines/chemokines associated with disease severity and adverse prognosis in COVID-19 patients
Authors: Cheng Jianghao , Wang Haozhen , Li Chaodan , Yu Jianhua , Zhu Mingli
Published: November 2024

Background

Cytokines and chemokines as crucial participants in innate immune response play significant roles during SARS-CoV-2 infection, yet excessive immune response exacerbates the severity of COVID-19.

Purpose

This study aims to investigate the involvement of which cytokines/chemokines in the cytokine storm of COVID-19, as well as the changes in cytokine/chemokine levels during the course of COVID-19, simultaneously exploring the diagnostic and prognostic value of the relevant cytokines/chemokines for COVID-19.

Methods

Flow cytometry was employed to detect the levels of cytokines and chemokines in the serum of 50 COVID-19 patients.

Results

Compared with severe COVID-19 patients, the levels of cytokines IL-6, IL-8, IL-10, sCD25, and chemokines IP-10 and MIG in the peripheral blood of non-severe patients were significantly reduced, while only IL-6, IL-10, and IP-10 levels were significantly decreased compared to non-survivors of COVID-19. Meanwhile, serum concentrations of IP-10, MCP-1, sTREM-1, IL-10, and the neutrophil-to-lymphocyte ratio (NLR) in peripheral blood could distinguish between COVID-19 survivors and non-survivors and were significantly associated with mortality. Among them, the concentration of IP-10 was shown to be the most powerful indicator for predicting adverse outcomes in COVID-19 patients (AUC: 0.715); however, its combined detection with the conventional inflammatory marker NLR did not improve the predictive value for adverse outcomes in COVID-19 patients. Additionally, serum IP-10 levels were negatively correlated with peripheral blood NK cell count and total lymphocyte count, while sTREM-1 levels were positively correlated with peripheral blood CD4+ T cell count and CD3+ T cell count. Meanwhile, IL-8 levels were positively correlated with total lymphocyte count in peripheral blood. Finally, the serum levels of cytokines/chemokines in non-survivors of COVID-19 increased significantly before death, while in survivors, they returned to normal levels before discharge.

Conclusions

Severely ill and non-surviving COVID-19 patients exhibit compromised immune function, with significantly higher levels of inflammation, cytokine/chemokine storms, and immune dysregulation compared to non-severe patients. Serum concentrations of IP-10, MCP-1, sTREM-1, and IL-10 levels can serve as biomarkers to predict adverse outcomes in COVID-19.
Associations between glycan signature alterations and the cellular antigenic properties of passaged chondrocytes
Authors: Homan Kentaro , Tokuhiro Taiki , Onodera Tomohiro , Hanamatsu Hisatoshi , Furukawa Jun-ichi , Ebata Taku , Matsuoka Masatake , Kadoya Ken , Terkawi M. Alaa , Iwasaki Norimasa
Published: November 2024

Background

Cartilage repair is a significant clinical challenge because of the limited intrinsic healing capacity. Current therapeutic strategies, such as cell transplantation therapy, aim to overcome this challenge by replacing damaged tissue with healthy cells. However, the long-term survival and functionality of transplanted cells remain major hurdles.

Objective

This study investigated the impact of chondrocyte passaging on glycan profiles and their antigenic properties. We hypothesized that alterations in glycan composition due to passaging may contribute to the enhanced ability to activate macrophages, thereby affecting the outcome of cell transplantation therapy.

Methods

Peritoneal macrophages and primary articular chondrocytes were isolated from C57BL/6 mice to establish direct and indirect coculture models. Macrophage activation was assessed by measuring the concentrations of IL-6 and nitric oxide in the culture supernatants or their gene expression. Glycome analysis of various glycoconjugates was performed by glycoblotting methods combined with the SALSA procedure for N-glycans and GSLs and the BEP method for O-glycans.

Results

Our results revealed that direct coculture of macrophages with passaged chondrocytes increased the production of proinflammatory cytokines, including IL-6 and NO, as the number of passages increased. With increasing passage number, the expression of GD3 substantially decreased, and the expression of GM3, especially GD1a, significantly increased. Coculturing passaged GM3S knockout chondrocytes with macrophages significantly suppressed IL-6 expression, implying reduced macrophage activation.

Conclusion

The observed activation of macrophages due to alterations in the glycan profile of chondrocytes provides a possible explanation for the antigenicity and immune rejection of transplanted cells.
Ferroptosis exacerbates the clonal deletion of virus-specific exhausted CD8+ T cells
Authors: Tian Qin , Chen Cheng , Lu Jinjin , Zheng Xinyu , Zhai Xiuming , Yang Yanping , Zhao Ziyao , Hao Jiangtao , Yang Ke , Ye Lilin , Wang Yifei
Published: November 2024

During chronic infection or tumorigenesis, persistent antigen stimulation contributes to the exhaustion of CD8+ T cells. Nevertheless, exhausted CD8+ T (TEX) cells still preserve certain effector function, and maintaining a reservoir of exhausted cells is of vital importance for virus elimination and tumor eradiation. Despite considerable work interrogating the rejuvenation of TEX cells, mechanisms underpinning the clonal deletion of TEX cells remain largely unexplored over the past decade. In this study, we employed mouse models of LCMV infection to demonstrate that excessive accumulation of lipid peroxidation rendered virus-specific TEX cells to ferroptosis, which may correlate with enhanced mitochondria-derived oxidative stress and compromised activity of glutathione peroxidase 4 (GPX4). In addition, either incomplete or complete ablation of GPX4 resulted in exacerbated ferroptosis and aggravated shrunken population of virus-specific TEX cells. On the other hand, inhibiting ferroptosis via administration of a ferroptosis inhibitor or overexpression of GPX4 greatly rectified the cell loss of virus-specific TEX cells. Collectively, we disclosed ferroptosis as a crucial player in the clonal deletion of virus-specific TEX cells and stressed the intervention of ferroptosis as a promising approach to optimize the longevity of virus-specific TEX cells.
Development of a pan-genotypic monoclonal antibody-based competitive ELISA for the detection of antibodies against Bovine viral diarrhea virus
Authors: Qi Shuhui , Wang Jing , Le Ting , Sun Chao , Chang Jitao , Jiang Zhigang , Yin Xin , Pang Quanhai
Published: November 2024

Introduction

Bovine viral diarrhea virus (BVDV), a positive-sense single-stranded RNA virus, causes significant economic losses in the cattle industry. Current diagnostic methods for BVDV exhibit variable sensitivity and specificity, underscoring the need for more rapid and accurate detection approaches. Here, we developed a novel competitive ELISA (cELISA) to detect antibodies against the BVDV E2 protein.

Methods and results

We generated three monoclonal antibodies (mAbs)—3E6, 2D5, and 5B9—by immunizing mice with purified BVDV E2 protein expressed in Expi293F cells. Among these, mAb 3E6 displayed superior competitive binding abilities to the E2 protein, enabling effective differentiation between BVDV positive and negative sera. Remarkably, mAb 3E6 exhibited pan-genotypic recognition of various BVDV strains, including BVDV-1a, -1b, -1c, -1m, -1p, -1v, and -2a, while showing no cross-reactivity with the classical swine fever virus (CSFV). Computational modeling using AlphaFold 3 identified domain B of the E2 protein as the primary binding site for mAb 3E6. Building upon these findings, we established a cELISA employing mAb 3E6 and recombinant E2 protein. Receiver-operating characteristic (ROC) analysis revealed outstanding diagnostic performance, achieving a sensitivity of 99.26% and specificity of 98.99%. Further tests confirmed the cELISA's specificity for detecting BVDV-specific antibodies, with no cross-reactivity with antisera from animals infected or immunized against BCoV, BHV-1, BRV, AKAV, LSDV, BLV, and CSFV. Consistency was observed between results from the BVDV E2 cELISA and traditional virus neutralization test (VNT), demonstrating high sensitivity for monitoring antibody dynamics. In performance evaluations, the established cELISA exhibited high concordance with VNT in assessing 160 vaccinated sera and 190 clinical samples.

Discussion

The BVDV E2 cELISA, utilizing mAb 3E6 to target domain B of the BVDV E2 protein, represents a reliable and effective serological diagnostic tool for the detection of antibodies against both BVDV-1 and BVDV-2. This methodology holds significant promise for applications in clinical diagnosis and the evaluation of vaccine efficacy.
13C-enrichment NMR spectroscopy: a tool to identify trophic markers and linkages
Authors: Bock Christian , Zhao Tianyi , Götze Sandra , Wermter Felizitas C. , Lannig Gisela
Published: November 2024

Current climate change, particularly ocean warming, will induce shifts in marine species distribution and composition, affecting the marine food web and, thus, trophic interactions. Analyses of the stable isotopes 13C and 15N are commonly used to detect trophic markers for food web analyses. With the current standard methods used in food web ecology, it is still challenging to identify potential changes in the uptake and utilization of trophic markers. In this work, we present a 13C-enrichment analysis by NMR spectroscopy to track the uptake and utilization of dietary carbon in a simple laboratory experiment of a primary producer and its consumer (algae and bivalve). In particular, we tested the hypothesis of a temperature-dependent use of dietary carbon by tracing the incorporation of 13C-atoms. Unicellular phytoplankton, Phaeodactilum tricornutum, was reared in a medium containing 13C-labeled bicarbonate. The accompanying 13C-NMR spectra of labeled P. tricornutum showed a specific profile of 13C-labeled compounds, including typical trophic markers such as the polyunsaturated omega-3 fatty acid eicosapentaenoic acid (EPA). Afterwards, 13C-labeled P. tricornutum was fed to King scallops, Pecten maximus, kept at two different temperatures (15°C and 20°C). Tissue-specific NMR spectra of P. maximus revealed elevated 13C-NMR signals, particularly of the fatty acid EPA in the digestive gland, which was not evident in muscle tissue. The comparison between the two temperatures indicated a change in trophic markers. At the higher temperature, less unsaturated fatty acids were detected in the digested gland, but increased 13C-labels in sugars were detected in the adductor muscle. This might indicate a change in the uptake and utilization of the trophic marker EPA in P. maximus due to a shift in energy conversion from favored beta-oxidation at colder temperatures to conversion from carbohydrates in the warmth. Our approach indicates that besides the accumulation of trophic markers, their incorporation and conversion are additional important factors for the reliable interpretation of trophic linkages under climate change.
Interspecific differences in growth, digestion, and feeding metabolism among freshwater fishes with different food habits
Authors: Pang Xu , Yang Jie , Xiang Shen-Ao , Sun Hao , Fu Shi-Jian
Published: November 2024

Studies of interspecific differences in the physiological performance of animals and their relationship with metabolism may provide insight into how selection influences diversity in phenotypic traits. To investigate the effects of food habits on growth, digestion, specific dynamic action (SDA), and the intestinal microbiota and verify their relationships in freshwater fishes, we measured the specific growth rate (SGR), meal remaining in the gastrointestinal tract, postprandial metabolic response, and intestinal microbial diversity of nine freshwater fish species with different food habits (three carnivorous, four omnivorous, and two herbivorous) fed at 25°C. The SGR, energy ingested (EI), energy efficiency (EE), gastrointestinal evacuation rate (GER), peak metabolic rate (PMR), peak metabolic scope (PMS), and SDA duration values were the highest in carnivores and the lowest in herbivores. The routine metabolic rate (RMR) and SDA coefficient values of carnivores were also significantly greater than those of omnivores and herbivores. The SGR was also significantly and positively correlated with the EI, GER, PMS, and SDA coefficients. These results indicated that growth performance, feeding and digestion abilities, and feeding metabolic responses were the strongest in carnivores and the weakest in herbivores. There was no significant difference in the richness of the intestinal microbiota with different food habits, whereas omnivores presented the highest diversity, followed by herbivores, and carnivores presented the lowest diversity. Moreover, while the phylum composition of the intestinal microbiota in fish with different food habits was similar, significant differences were observed in the composition proportions among carnivores. This might be related to the host’s ability to digest and utilize proteins, fats, and carbohydrates. We suggest that the interspecific differences in growth associated with different food habits are related to their feeding and digestive abilities, metabolic response, and gut microbiota.
Reevaluating 30 cmH2O endotracheal tube cuff pressure: risks of airway mucosal damage during prolonged mechanical ventilation
Authors: Mu Guo , Wang Feixiang , Li Qiang , Yu Xuan , Lu Bin
Published: November 2024

Background

The optimal endotracheal tube (ETT) cuff pressure remains contentious. In the traditional consideration that the level 30 cmH2O is considered safe, balancing the prevention of reflux aspiration against airway mucosal damage. Whether this pressure level can cause potential damage to the airway mucosa remains to be discussed.

Methods

Airway mucosa damage and structural changes at 30 cmH2O were examined in patients under general anesthesia and in rabbit mechanical ventilation models. Prior to this, we also interviewed some anesthesiologists about the level of concern about ETT cuff pressure.

Results

A total of 634 valid questionnaires suggested that anesthesiologists generally do not pay enough attention to ETT cuff pressure and the average established cuff pressure significantly exceeded 30 cmH2O. Airway mucosa images of 100 general anesthesia patients with different ventilation duration indicated that maintaining the pressure at 30 cmH2O did not cause significant damage to airway mucosa in a short period of time, while it still caused damage to airway mucosa in patients with long-term ventilation, with damage severity increasing with longer ventilation periods. This correlated strongly with postoperative sore throat (R2 = 0.3884, p < 0.001). In rabbits, 4 h of ventilation at this pressure resulted in significant loss of ciliated epithelium and inflammation. Calculations suggested an effective dose (ED50) to prevent mucosal injury at a cuff pressure of 25.64 cmH2O (95% CI: 19.268–29.367 cmH2O).

Conclusion

The currently established cuff pressure of 30 cmH2O is associated with airway mucosal damage in both clinical and animal models. Lowering the safety threshold of cuff pressure may be necessary to mitigate mucosal injury.
Progress in the mechanism of functional dyspepsia: roles of mitochondrial autophagy in duodenal abnormalities
Authors: Zhong Kexin , Du Xiaojuan , Niu Yuanyuan , Li Zhengju , Tao Yongbiao , Wu Yuqian , Zhang Ruiting , Guo Linjing , Bi Yurong , Tang Lijuan , Dou Tianyu , Wang Longde
Published: November 2024

Mitochondria are the main source of energy for cellular activity. Their functional damage or deficiency leads to cellular deterioration, which in turn triggers autophagic reactions. Taking mitochondrial autophagy as a starting point, the present review explored the mechanisms of duodenal abnormalities in detail, including mucosal barrier damage, release of inflammatory factors, and disruption of intracellular signal transduction. We summarized the key roles of mitochondrial autophagy in the abnormal development of the duodenum and examined the in-depth physiological and pathological mechanisms involved, providing a comprehensive theoretical basis for understanding the pathogenesis of functional dyspepsia. At present, it has been confirmed that an increase in the eosinophil count and mast cell degranulation in the duodenum can trigger visceral hypersensitive reactions and cause gastrointestinal motility disorders. In the future, it is necessary to continue exploring the molecular mechanisms and signaling pathways of mitochondrial autophagy in duodenal abnormalities. A deeper understanding of mitochondrial autophagy provides important references for developing treatment strategies for functional dyspepsia, thereby improving clinical efficacy and patient quality of life.
Erucic acid utilization by Lactobacillus johnsonii N6.2
Authors: Thompson Sharon C. , Beliakoff Reagan , Garrett Timothy J. , Gonzalez Claudio F. , Lorca Graciela L.
Published: November 2024

A multivariate nutritional analysis indicated that the consumption of erucic acid-rich food, a fatty acid (FA) found primarily in rapeseed and mustard oil, was positively correlated with higher counts of lactic acid bacteria (LAB). Furthermore, we showed Lactobacillus johnsonii N6.2, as well as other species of LAB tested from the former Lactobacillus genus, were able to efficiently use erucic acid (EA) as the source of FA. In this work, we identified significant changes induced in the FA profiles of L. johnsonii cultured with EA as the source of FA. We performed global transcriptomics to identify genes and pathways involved in EA utilization. It was found that L. johnsonii incorporates external fatty acids via a FakA/FakB and the plsX/plsY/plsC pathway for phosphatidic acid synthesis. It was found that cells grown in MRS with EA (MRS-E) significantly upregulated fakB2 and fakB4 when compared to cells grown in standard MRS with tween 80 as the source of FA. Additionally, in MRS-E, L. johnsonii N6.2 induced the expression of plsY2, plsC2 and plsC4 while the expression of pslX was constitutive during short term EA exposure. LC–MS analyses revealed that L. johnsonii N6.2 rapidly incorporates EA and synthesizes a variety of long chain fatty acids, including the health-relevant omega-9 monounsaturated fatty acids such as nervonic and gondoic acids.