bluuebunny/crossref_metadata_2025 huggingface dataset:
TGF-β signaling limits effector function capacity of NK cell anti-cancer immunity in human bladder cancer
Authors: Wong Joshua , Rodrigues Rossi Gustavo , Guimaraes Fernando
Published: November 2024

NK cells are important innate immune cells that have unique abilities to recognize and eliminate cancer cells, particularly through antibody dependent cellular cytotoxicity (ADCC). However, NK cell-based responses in bladder cancers to targeted antibody therapeutics are typically immunosuppressed, and these mechanisms in this disease are largely unknown. Single cell RNA sequencing (scRNA-seq) and high-dimensional flow cytometry were used to investigate the phenotype of tumor-infiltrating NK cells in samples from patients with bladder cancer. NK cells within bladder cancer displayed reduced expression of FcγRIIIa/CD16, the critical Fc receptor involved in ADCC-mediated cytotoxicity, on both transcriptional and protein levels. Transcriptional signatures of transforming growth factor (TGF)-β-signaling, a pleiotropic cytokine known for its immunosuppressive effects, were upregulated in tumor-infiltrating NK cells. We further validated TGF-β mediated CD16 downregulation on NK cells in vitro, which was accompanied by a transition into a tissue residency phenotype. CD16 downregulation was also abrogated by TGF-βR signaling inhibition, which could also restore the ADCC ability of NK cells subject to TGF-β effects. This study highlights how TGF-β-rich bladder cancers inhibit NK cell-mediated ADCC by downregulating CD16. TGF-β inhibition represents new avenues to reverse immunosuppression and enhance the tumoricidal capacity of NK cells.
Natural killer cells display unique, non-cytotoxic profiles in human endometrial cancer
Authors: Eisele Caprice , Riedinger Courtney , Haight Paulina , Kowalski Jesse , Mundy-Bosse Bethany , Porrett Paige , Collins Patrick , Cosgrove Casey , Freud Aharon
Published: November 2024

Endometrial cancer (EC) responds variably to immune checkpoint blockade, warranting study of the tumor microenvironment (TME). Natural killer (NK) cells are cytotoxic innate lymphocytes that can directly kill tumor cells, yet their diversity, locations, and functions in the EC TME are unknown. CITE-seq analysis of fresh EC tumor samples revealed multiple NK cell populations in the TME, with distinct patterns of surface activating and inhibitory receptor expression and less cytolytic granule expression by tumor-derived conventional NK (cNK) and tissue-resident NK (trNK) cells compared to autologous blood cNK. Ex vivo flow cytometry revealed significantly lower expression of cytolytic granules by TME-derived NK cell subsets (granzyme B MFI: blood cNK 1.5e5 +/- 5.4e4, tumor cNK 7.9e4 +/- 2.9e4 p<0.05, tumor trNK 3.7e4 +/- 1.4e4 p<0.05, n=10; perforin MFI: blood cNK 1.3e4 +/- 3.1e3, tumor cNK 1.0e4 +/- 3.2e3 p=0.19, tumor trNK 3.5e3 +/-1.3e3 p<0.01, n=8). To identify NK cell subsets within the TME, we performed immunohistochemistry using CD3, CD56, granzyme B, and CD103 to demarcate cNK and trNK, respectively. We observed granzyme B+ cells primarily in the stroma (78.6 +/- 3.7% Stroma, 21.4 +/- 3.7% Glands, n=10, p<0.05) and CD103+ cells primarily among tumor cells (15.6 +/- 4.5% Stroma, 84.3 +/- 0.64% Glands, n=10, p=0.07). These data suggest that both cNK and trNK cell cytotoxicity is suppressed in the EC TME and ongoing work seeks to elucidate the mechanisms.
Role of the Afferent Lymph as an Immunological Conduit to Analyze Tissue Antigenic and Inflammatory Load
Authors: Nanaware Padma , Clement Cristina , Khan Zohaib , Mota Ines , Stern Lawrence , Santambrogio Laura
Published: November 2024

The lymphatic fluid is the conduit by which part of the tissue “omic’ is transported to the draining lymph node for immunosurveillance. Following cannulation of the pre-nodal cervical and mesenteric afferent lymphatics we analyzed the lymph proteomic composition and determined that its tissue specificity is reflected in the dendritic cell-MHCII-eluted immunopeptidome harvested from the cervical or mesenteric nodes respectively. Additionally, since the mesenteric lymph drains directly from the lymphatics underlying the intestinal barrier, we performed the same analysis following inflammatory disruption of the gut barrier to map the antigenic and inflammatory load draining to the mesenteric lymph nodes, in both mice and human subjects with inflammatory bowel diseases. The gastrointestinal tissue damage was reflected in microbiome-derived by-products, components of the bacterial wall and other immunomodulators, including molecules of the gut-brain axis, found in the afferent lymph following damage to the gut epithelium. Our data points to the relevance of the lymphatic fluid to probe the tissue-specific antigenic and inflammatory load transported to the draining lymph node for immunosurveillance.
PD-1/cDC1 axis in the tumor draining lymph node regulates stem-like CD8+ T cell differentiation
Authors: Hor Jyh Liang , Schrom Edward , Germain Ronald
Published: November 2024

Stem-like CD8+ T cells constitute the major proliferative cytotoxic CD8+ T cell subset during checkpoint therapy. While it is increasingly appreciated that tumor draining LNs (tdLN) serve as reservoirs from which intratumoral stem-like CD8+ T cells can be replenished, how activated CD8+ T cells interact with the tdLN environment and differentiate into stem-like T cells remain unresolved.

Using multiplexed 3D tissue microscopy, we identified that during late differentiation (Days 4-8+), TCF-1+ PD-1+ stem-like CD8+ T cells formed distinct clustering niches with XCR1+ dendritic cells (cDC1), which serve as sites of ongoing antigen presentation in the tdLN that drive continuous stem-like T cell expansion. Intriguingly, PD-1 expression among polyclonal TCF-1+ stem-like cells is positively correlated with tetramer binding, suggesting that the PD-1/cDC1 axis promotes selective enrichment and expansion of high affinity stem-like cells during effector differentiation. In vivo blockade of PD-1/PD-L1 signaling resulted in the loss of high affinity stem-like clones in the tdLN, along with a phenotypic shift towards terminally differentiated effector cells.

Our findings raise questions about whether prolonged use of PD-1 checkpoint therapy in cancer patients interferes with maintenance of a key precursor population necessary for the robust effector generation needed to achieve full tumor regression.

This work was supported by the Intramural Research program of NIAID, NIH.
Targeting Pim2 improves effector function and longevity of T cells in cancer immunotherapy through regulating autophagy
Authors: Wu Yongxia , Tian Linlu , Pugel Allison , Yu Xue-Zhong
Published: November 2024

The Pim kinase family is critically involved in tumorigenesis, yet its role in primary T cells is not well known. We reported that Pim2, distinct from the other two isoforms, inhibits T-cell responses to alloantigen. To study how Pim2 impacts T-cell immunity against cancer, we used murine models of adoptive cell therapy (ACT) and hematopoietic cell transplantation. Pim2 deficiency enhanced the ability of antigen specific CD8 T cells and polyclonal T cells in controlling solid tumor and leukemia growth, respectively. Pim2 deficiency increased cytokine production and metabolic activities of CD8 T cells in vitro and promoted effector but constrained exhausted CD8 subsets in tumor. Pim2-/- CD8 T cells had increased Tcf1 expression and longevity after ACT. Mechanistically, Pim2 binds P62 and constrains T-cell activation through facilitating autophagy. Pim2 deficiency in T cells attenuated LC3 lipidation, P62 degradation as well as autophagosome formation. Augmentation of autophagy via Atg5 overexpression or metformin treatment reduced effector differentiation and anti-tumor function of Pim2-/- T cells. Importantly, a Pim2-specific inhibitor improved murine T-cell immunity against melanoma and leukemia and increased activity of human melanoma-specific T cells and CD19 CAR-T cells. Taken together, Pim2 restrains T-cell anti-tumor immunity through regulating autophagy, and blocking Pim2 enhances effector differentiation and persistence of T cells after cancer immunotherapy.
RNA sequencing methods enable immune cell gene expression profiling and clonality determination
Authors: Song Chen , Janzen Evan , Devoe Tessa , Erijman Ariel , Naishadham Gautam , Langhorst Bradley
Published: November 2024

The adaptive immune system defends the human body from pathogens through B/T cell functions in various organs. Abnormality of these cells may reflect immunological disorder, presence of neoplastic tissue, or response to infectious agents. High-throughput sequencing technology has facilitated understanding gene expression and related cell functions. Still, an in-depth understanding of B and T cell phenotype and clonotype profiling is an emerging application that has yet to take full advantage of sequencing technology for disease diagnosis and prognosis.

Here, we share results of RNA sequencing of immune system tissues, including healthy donor peripheral blood mononuclear cells (PBMC), lymph nodes, bone marrow, spleen, thyroid, thymus, and colon, as well as diseased PBMC and colon samples. Sequencing libraries were constructed and sequenced for B and T cell-specific gene expression and full-length immune repertoire profiling. We have identified tissue-specific B/T cell phenotype and clonality patterns related to tissue functions and disease progression. RNA-seq enables highly multiplex immunophenotyping and clonality determination for high-abundant clones that correlate with Immune-seq, with the latter targeted approach providing more in-depth clonality signatures. Integrating both RNA sequencing and immune repertoire sequencing approaches allows accurate phenotype and clonotype determination for the immune landscape in various tissues.
Multiplexed ImmunoSpot™ enables streamlined assessment of antigen-specific B cells; lessons toward maximizing cell utilization and increasing sample throughput
Authors: Kirchenbaum Greg , Chepke Jack , Becza Noemi , Yao Lingling , Liu Zhigang , Lehmann Paul
Published: November 2024

There are few techniques capable of detecting rare antigen-specific B cells while also providing information regarding their abundance, Ig class/subclass usage and functional affinity. The ImmunoSpot™ assay approach is suited for such antigen-specific B cell assessments since it provides information at single-cell resolution for individual ASC. Moreover, recently introduced innovations in the implementation of this assay modality have opened the door to testing larger sample sets. Towards this objective, multiplexed B cell FluoroSpot assays reduce the cell material requirement through parallel assessment of multiple Ig classes or IgG subclasses. Nevertheless, accurate determination of ASC frequencies, especially in the context of diverse antigen panels and complex exposure histories, is best achieved through evaluating multiple cell inputs to accommodate the inherent variability present within and amongst individual donors. The scope of this research effort was to further optimize the assay pipeline to facilitate high-throughput suitable assessment of antigen-specific ASC. This goal has been achieved through introducing advancements in the image analysis software component, improving both the time requirement and precision of downstream data analysis. Collectively, implementation of these assay optimizations serves to increase the feasibility of making high-throughput assessment of antigen-specific ASC using the ImmunoSpot™ approach a routine practice in immune monitoring.
Integration of single-cell transcriptomic and chromatin accessibility on heterogenicity of human peripheral blood mononuclear cells utilizing microwell-based single-cell partitioning technology
Authors: Hatami Elham , Song Hye-Won , Huang Hongduan , Zhang Zhiqi , McCarthy Thomas , Kim Youngsook , Li Ruifang , Bao Quyen , Nguyen Rosary , Narayan Punya , Wang Larry , Ayer Aruna
Published: November 2024

Single-cell RNA sequencing (scRNA-Seq) deepens our understanding of cellular development and heterogeneity. However, limitations exist in unraveling cell states and gene regulatory programs. Chromatin state profiles assess gene expression potential and offer insights into transcriptional regulation. Integrated with gene expression data, chromatin accessibility region (CAR) profiles establish fundamental gene regulatory logic for cell fate. ATAC-seq (Assay for Transposase-Accessible Chromatin using Sequencing) is a highly potent approach for profiling genome-wide CARs.

To investigate the power of the multiomics assay in identifying differentiated gene regulations, we conducted multiomic snATAC-seq+ snRNA-seq on PBMCs from two different donors, by utilizing the gentle and robust microwell-based single-cell partitioning technology. The assay showed high sensitivity and specificity metrics (>10,000 median unique fragments/cell, >0.7 fragments in peak score). Integrative analysis across donors revealed enriched transcription factor motifs and fragment coverage tracks in distinct cell types, correlating significantly with gene expression data. These findings highlight mRNA's intricate connections with CARs in immune cell development.

Our study underscores the power of multiomic analysis in analyzing heterogeneity of PBMC cell populations and offers a toolkit to identify gene regulation specific to diverse cell types, enhancing our comprehension of epigenetic diversity.
Next-Level Efficiency: StraightFrom Spleen Isolation Kits - Standardizing Isolation of Untouched T cells with Minimal Hands-On Time
Authors: Kostadinovska Konstantina , Flade Sebastian , Borczewski Amelie , Neufeld Elina , Dhana Ermanila , Winkels Gregor , Baranska Anna
Published: November 2024

Isolation of T cells from mouse spleen is essential for various research fields. Studies using magnetically separated cells help to understand the immune system, develop new vaccines and advance immunotherapy. Thus, selecting a reliable cell isolation method is critical for consistent experimental results.

Simultaneous spleen dissociation and labeling of unwanted cells with an antibody cocktail was performed using the GentleMACS dissociator. Subsequent automated magnetic labeling of cells and isolation of untouched T cells (CD4, CD8 or Pan T) was performed using the autoMACS NEO Separator. Target cell purity, yield, cell activation status and functionality were assessed.

Untouched CD4, CD8 or Pan T cells obtained with our StraightFrom Spleen workflow were of the same high purity and yield as conventional more time-consuming protocols, which require preparation of single cell suspension and various centrifugation steps prior to enrichment. Flow cytometric analysis of isolated cells revealed no expression of the activation markers CD25 or CD69 in non-stimulated cells even after overnight cultivation. However, significant upregulation was observed after stimulation. In addition, isolated cells showed excellent proliferation in culture.

We have developed a simple and fast cell isolation method that significantly accelerates the isolation of functional cells, providing an attractive alternative to the laborious multi-step protocols used in academic and industrial laboratories.
Molecular Pixelation: Spatial proteomics of single-cells by next generation sequencing
Authors: Glavas Steven
Published: November 2024

Molecular Pixelation (MPX) is a novel single cell spatial proteomics panel able to simultaneously quantify protein abundance, the spatial distribution, and colocalization of targeted cell surface proteins on up to 1,000 individual cells.

MPX creates 3D spatial maps of cells by imprinting spatial information on antibody oligonucleotide conjugates using a DNA reagent we call DNA-pixels. These DNA-pixels form over 1,000 connected spatial zones per single cell producing graphs that reconstruct the cell surface in silico; forming a single cell surface map of 80 proteins. By applying spatial statistics on these cell surface graph representations, we uncover both known and novel patterns of protein spatial polarization and co-localization associated with vital immune processes such as intercellular communication and mobility.

In our recent paper we show how MPX can be used to monitor the cell surface effects of treatment or stimulation, such as the spatial restructuring of the T cell surface proteins upon the formation of the uropod, the hind part of a migrating T cell.

With high throughput and multiplex, MPX is a powerful tool for investigating the intricate spatial organization of proteins in single cells, paving the way for new discoveries in immune system research.
Making a stabilized HLA-E complex with native properties.
Authors: Blicher Thomas , Mortensen Tobias , Brix Liselotte
Published: November 2024

Objective

The non-classical, human MHC-I molecule HLA-E is functionally monomorphic and therefore a highly attractive target in immunotherapy against cancers and infectious diseases. However, HLA-E is characterized by low stability due to its biological role as a rapid-response system to monitor HLA-I expression. The inherent instability makes production of HLA-E difficult and severely limits its use as a reagent, which in turn negatively impacts the understanding of HLA-E biology. Here, we present native-like, stabilized forms of HLA-E, which brings production of HLA-E in line with that of classical MHC-I molecules.

Methods

Disulfide bridges were introduced in HLA-E at selected locations deemed to not interfere with recognition of HLA-E by NK or T cell receptors. The stability of the modified HLA-E molecules was monitored using thermal denaturation, and conservation of native HLA-E properties was assessed by in vitro TCR-binding assays and through staining of NK cells.

Results

The modified HLA-E molecules exhibited increased stability, increased refolding yields, both or neither. The most stable HLA-E variant exhibited thermal stability comparable to complexes of HLA-A*02:01, while retaining native-like recognition by both NK and T cell receptors.

Summary

Stabilized forms of HLA-E have been generated and their native-like properties validated using in vitro assays and NK cell staining. This enables production of reagents for the study HLA-E biology.
Profiling antigen-reactive T cells using high throughput single-cell T cell receptor sequencing.
Authors: Papalexi Efthymia , Hariadi Bryan , Curca Crina , Roco Charles , Rosenberg Alexander
Published: November 2024

Recent advances in single-cell sequencing allow for simultaneous profiling of T cell receptors (TCRs) and transcriptomes leading to the characterization of key T cell subsets with antigen recognition and killing capabilities. Despite their success, these methods rely on microfluidic devices or plate-based protocols with limited sensitivity and throughput (1,000s-10,000s of cells, 1-2 samples) making the study of disease-relevant T cells time consuming and costly.

To overcome these limitations, we developed a split pool combinatorial barcoding technology to simultaneously characterize the TCRs alongside the full transcriptomes of up to 1 million T cells from up to 96 unique samples. Here, we profiled hundreds of thousands of human T cells stimulated with antigens from a variety of diseases. Applying our method, we performed deep characterization of antigen-reactive TCR clonotypes and assessed their sequence similarities. In addition, we performed transcriptome-based clustering analyses and identified highly proliferative and cytotoxic T cell subsets. Finally, we correlated T cell activation and cytotoxicity states to clonal expansion frequencies in order to identify TCRs with increased killing potential.

In summary, we present a highly flexible and scalable combinatorial barcoding technology to allow researchers to profile up to 1 million T cells in one experiment and investigate their functional responses during infection, cancer, autoimmunity, or therapeutic interventions.
VERLO: a novel imaging cell sorter for enhanced analysis of tumor infiltration by immune cells
Authors: Dabelic Rea , Munoz Adonary , George Micayla , Xia Lin , Tang Rui
Published: November 2024

Cancer ranks as the second leading cause of death in the US. Central to cancer progression is the tumor microenvironment (TME), which significantly influences the disease's spread. Immune cell infiltration into the TME is crucial for effective immunotherapy. Traditional infiltration assessment methods, including microscopy and flow cytometry, have limitations such as low throughput and lack of imaging. Addressing these issues, we present VERLO, an imaging cell sorter combining low-pressure microfluidics with dual-laser scanning to enhance cell analysis. We utilized VERLO to examine Jurkat cells, a T-cell proxy, infiltrating breast cancer spheroids, which simulate tumors. VERLO's application enabled us to preserve Jurkat-tumor interactions, facilitating the exploration of factors that may augment immune infiltration. This research underscores VERLO's utility in studying immune cell dynamics within the TME, advancing immunotherapy optimization for cancer treatment. Our findings indicate that VERLO can significantly contribute to understanding and improving immune cell-mediated cancer therapies.
HLA-A2 Chimeric antigen receptor regulatory T cells promote allograft acceptance in mice
Authors: Durgam Samarth , Sayin Ismail , Hynes Grace , Yin Dengping , Rosado sanchez isaac , Speck Madeleine , Mojibian majid , Levings Megan , Chong Anita
Published: November 2024

Purpose: Regulatory T cells (Tregs) are immune regulators essential for achieving stable allograft acceptance. We tested whether HLA-A2 CAR-Tregs have the therapeutic potential to prolong allograft survival through mediating linked-suppression and infectious tolerance. Methods:Donor hearts expressing BALB/c, HLA-A2 and 2W-OVA antigens (C57BL/6.A2 X BALB/c.2W-OVA.F1) were transplanted into C57BL/6 mice and received 250µg aCD154 ± 1X106 A2 CAR-Tregs on the day of transplant (HTx). 1) Graft survival was monitored. 2) CAR-Tregs in the peripheral circulation, spleen and lymph nodes (SLO) were monitored (Thy1.1+FoxP3+). 3) 2W-specific Tregs and Tconvs quantified at ~D-45 post-HTx. 4) Donor specific IgG was quantified. Results: 1)HLA.A2-CAR Tregs + 250µg aCD154 significantly prolonged allograft survival compared to controls with 250µg aCD154 alone (Mean: >56 vs 26 days). 2)FoxP3+CAR-Tregs were detected in the circulation at D10-40 post-HTx, and in the SLO at >D45. 3) In the SLO, CAR-Tregs mediated linked suppression by preventing 2W-specific Tconvs expansion, and infectious tolerance by expanding of 2W-specific Tregs (7-15-fold). 4)A2.CAR-Tregs significantly decreased anti-A2 and anti-donor MHCI & MHCII IgG production. Conclusion: HLA-A2.CAR-Tregs synergized with aCD154, mediated linked suppression and infectious T cell tolerance and significantly prolonged allograft survival. Remarkably, HLA-A2.CAR-Tregs suppressed donor MHC-specific IgG responses and anti-HLA-A2IgG.
Multi-omic immunophenotyping to identify predictors of mRNA vaccine responsiveness in immunocompromised individuals
Authors: Murray Sam , Meacham Georgina , Irwin Sophie , Rao Vishal , Rolt Jordan , Dimitriadis Stavros , Dooley Kyla , Heslington Amelia , Goodyear Carl , Siebert Stefan , McInnes Iain , Willicombe Michelle , Thomas David , Billingham Lucinda , Kearns Pamela , Marjot Thomas , Kronsteiner Barbara
Published: November 2024

Recent advances in -omics technologies enable detailed evaluation of the human immune system in the context of important clinical phenotypes. Here, we apply such technologies to investigate predictors of vaccine immune responsiveness in individuals with a range of immunosuppressive conditions who are vulnerable to COVID-19 vaccine failure.

Using bulk and single-cell RNA sequencing (scRNA-seq), Olink proteomics and spectral flow cytometry we assessed the pre- and 21 days post-third dose COVID-19 mRNA vaccine immunophenotype of 133 immunocompromised individuals and 22 healthy controls. Machine learning dependent analyses associated these phenotypes with vaccine-induced antibody and T cell responses.

We identified distinct pre-vaccination transcriptional signatures which predicted post-vaccine antibody responsiveness across immunosuppressive conditions. Signatures included increased B cell related gene modules and decreased inflammatory and monocyte related modules. These patterns were confirmed by detailed cellular and proteomic immunophenotyping. Persistence of this altered phenotype after vaccination was observed using scRNA-seq, which gave insights into the cellular identity of signatures related to vaccine responsiveness.

Through comprehensive immune profiling of pre- and post-vaccine blood samples from immunocompromised individuals we identify novel cellular and molecular signatures that predict vaccine responsiveness, and give insight into mechanisms of vaccine failure.
HIV vaccine candidate efficacy mediated by epigenetic changes in CD14+ cells, cAMP-dependent efferocytosis and immunoinhibitory DC-10
Authors: Bissa Massimiliano , Silva de Castro Isabela , Kim Sohyoung , Fourati Slim , Rahman Mohammad Arif , Tomalka Jeffrey , Schifanella Luca , Sarkis Sarkis , Gutowska Anna , Sekaly Rafick , Hager Gordon , Franchini Genoveffa
Published: November 2024

The V1-deleted DNA/ALVAC/gp120 anti-HIV vaccine significantly decreased the risk of SIV and SHIV acquisition in macaques. CD14+ monocytes and the Antibody-dependent cellular cytotoxicity to V2 (V2-ADCC) were correlated with reduced risk, indicating the central role of innate response in mounting an effective response. We hypothesized that in-depth characterization of the innate responses to vaccination will elucidate mechanisms of protection.

To test this hypothesis, we integrated different analyses conducted in multiple macaque studies to investigate how the innate and adaptive responses cooperate in reducing the acquisition. Analyses of samples collected from V1-delted DNA/ALVAC/gp120 vaccinated macaques included canonical and multi-omics assays.

The study of the epigenetic landscape of CD14+ cells showed that epigenetic reprogramming of enhancer region upstream of CREB1 gene was associated with reduced risk of infection. Furthermore, we found that, the cyclic AMP (cAMP)-dependent efferocytosis performed by CD14+ monocytes that engulfed and clear apoptotic cells, correlated with reduced risk of acquisition and complemented V2-ADCC. Finally, we identified a crucial role of immunoinhibitory dendritic cells DC-10 in lowering the risk of infection.

These data suggest that V2-ADCC, efferocytosis and immunoinhibitory innate cells contribute to vaccine efficacy by decreasing inflammation and maintaining tissue homeostasis, and therefore counteract the SIV/SHIV infection.
Unveiling the immune responses of ferrets inoculated with Ebola virus
Authors: Bernbaum-Cutler Rebecca , Marketon Anthony , Rivera Deja , Gale Eric , Mani Venkatesh , Anthony Scott
Published: November 2024

Animal-based models continue to be important for investigating the immunological pathways involved with infections of emerging infectious diseases. Inoculation of Ebola virus (EBOV) in ferrets results in nearly uniformly fatal disease while also recapitulating numerous additional aspects associated with EBOV infection of nonhuman primates or humans. However, the underlying immune responses of EBOV-associated disease in ferrets is largely unknown due to a lack of available reagents. Thus, we designed and optimized a 10-color flow cytometry panel to investigate the adaptive and innate immune components of ferrets. Animals were inoculated with EBOV or mock virus, and circulating immune responses in whole blood were longitudinally examined by flow cytometry. Appreciable alterations in both the innate and adaptive immune responses during late-stage disease were observed. Animals were euthanized at 3 or 6-d post-exposure, and terminal tissues were examined. As expected, marked increases in the number and frequency of apoptotic and necrotic cells were noted. Interestingly, a significant loss of memory phenotype CD8+ T cells and activated CD11b+ cells was associated with advanced disease. In plasma, elevated circulating protein levels of multiple cytokines, commonly associated with EBOV infection, were found in a stepwise manner. Altogether, these tools add significant illumination of underlying immune responses in the ferret model.
Management of fever and associated symptoms in children and adults: an Italian national survey
Authors: Chiaretti Antonio , Grattagliano Ignazio , Carlomagno Francesco , Magi Massimo , Carbone Luigi , Zecca Silvia , Bondone Claudia , Gatto Antonio , Di Sarno Lorenzo , Franceschi Francesco
Published: November 2024

An advisory committee of Italian experts conducted a national, cross-sectional, web-based survey in June 2024 to describe the current management of fever and associated symptoms in different settings, including primary care, emergency departments, and hospital wards in pediatric and adult patients. The survey covered two domains: participants’ features and questions about the main drugs prescribed to treat fever and associated symptoms. A total of 832 questionnaires were analyzed. Paracetamol was the most prescribed drug to treat fever. Most participants were influenced by related symptoms when choosing the most appropriate drug. Almost all participants selected the oral route as their preferred one. This survey provides a current state of fever management among physicians in different settings throughout Italy. It highlights a trend in treating fever with antipyretics at appropriate doses and evaluating pain through validated scales.
I diritti dell'embrione
Authors: Leone Salvio
Published: November 2024

Not available
Making Alexander von Humboldt's works known in Lombardy: the American volumes of Giulio Ferrario's Costume antico e moderno
Authors: Banfi Enrico , Bianconi Filippo , Domenici Davide , Visconti Agnese
Published: November 2024

It is well known that the Braidense Library in Milan was one of the few, perhaps the only one, of the Italian peninsula to have in good time purchased the volumes of the Voyage aux Régions Équinoxiales du Nouveau Continent, fait en 1799, 1800, 1802, 1803 et 1804 (Paris, 1835-1834) that the famous Prussian naturalist-explorer Alexander von Humboldt (1769-1859) was gradually writing after returning from his expedition to Central and South America (1799-1804)....
Africa’s public service delivery and performance overview 2024
Authors: Simatele Munacinga C.
Published: November 2024

No abstract available.
Disaster preparedness: The role of spatial disaster learning using geospatial technology
Authors: Handoyo Budi , Soekamto Hadi , Putra Alfyananda K. , Kamil Puspita A. , Wulandari Fajar
Published: November 2024

Preparedness for natural disasters is crucial for people living in high-risk areas along the Ring of Fire, such as people in Indonesia. Although Spatial Disaster Learning using Geospatial Technology (SDL-GeoTech) has been developed, its effectiveness in enhancing students’ preparedness remains unproven. This study examines the impact of SDL-GeoTech on junior high school students’ readiness using a quasi-experimental, time-series, single-group pre-post-test design. The research involved students from three schools: SMA 1 Grati Pasuruan (flood-prone), SMA 1 Puncu Kediri (volcanic eruption-prone), and SMA 3 Mataram Lombok (earthquake-prone). Data were collected through tests on knowledge, skills, and attitudes, followed by t-test analysis at a 0.05 significance level. The results of this study show SDL-GeoTech was significantly able to improve students’ preparedness, including their knowledge, skills, and attitudes. These findings highlight SDL-GeoTech as an innovative tool for disaster education, with the potential to enhance curricula and teaching strategies, especially in vulnerable areas.Contribution: This research teaches students to use techno-geospatial learning through SDL-GeoTech, equipping those in the Ring of Fire region to be better prepared for potential natural disasters such as earthquakes and floods. The proven analysis of SDL-GeoTech has shown that it enhances students’ knowledge, skills and attitude in preparedness in dealing with disasters.
Enhancing emotional intelligence for well-being in higher education: Supporting SDG 3 amid adversity
Authors: Maharaj Prashanti , Ramsaroop Anisha
Published: November 2024

Orientation: The coronavirus disease 2019 necessitated tertiary institutions to reevaluate their educational paradigms, emphasising resilience and fostering creative mindsets to navigate unprecedented challenges. This disruption evoked distressing emotions, posing threats to academics’ emotional well-being. Such challenges carry profound implications for the attainment of sustainable development goal 3 (SDG 3), aimed at ensuring healthy lives and promoting well-being for all.Research purpose: This study aimed to comprehend and harness the emotional intelligence of academics, with a view to understanding how adversity affects emotional intelligence and emotional well-being in academics.Motivation for the study: The focus on enhancing the emotional well-being of academics directly contributes to the overall health and well-being goals outlined in SDG 3.Research approach/design and method: A quantitative approach with a sample size of 360 academics was drawn from five campus sites from the University of KwaZulu-Natal using stratified random sampling.Main findings: Respondents reported elevated levels of stress and anxiety and a lack of visibility regarding employee assistance programmes (EAPs). Respondents indicated a notable dissatisfaction with the lack of support from the human resource department.Practical/managerial implications: Recommendations proposed in the higher education landscape for advancing SDG 3 consisted of EAP insights and awareness, needs analysis on well-being support and policy formulation.Contribution/value-add: By focussing on emotional intelligence, the study aligned with SDG 3 and presented practical strategies embedded in a model designed to enhance the emotional well-being of academics.
Navigating the neoliberal university: A collaborative autoethnography of emerging scholars
Authors: Shabalala Nokulunga , Mapaling Curwyn
Published: November 2024

In the dynamic landscape of the neoliberal university, conversations between emerging scholars serve as vital spaces for critical reflection and transformative action. This collaborative autoethnographic study engaged with the complexities of navigating academia as two black clinical psychologists within a South African university. Drawing on decoloniality, we interrogated the pervasive ‘carrying on’ culture and its impact on early-career academics. Our lived experiences underscored the intersections of identity, power and resistance, as we grappled with the commodification of higher education and the pressures to ascend the ranks hastily. Through a reflexive thematic analysis of our recorded discussions, we uncovered mechanisms for disrupting normative structures and redefining the purpose of scholarly pursuits. Central to our inquiry was the notion of refusal as a generative force, challenging the status quo and advocating for a more conducive, supportive environment where teaching and learning activities are genuine expressions of growth. We envisioned a university that fosters meaningful intellectual engagement and societal transformation, calling for collective dialogue and action to reimagine the neoliberal higher education landscape.Contribution: Our study contributes to the ongoing conversation on decolonising academia, offering insights into the struggles and aspirations of emerging scholars in the Global South, and advocating for a transformative praxis that nurtures authentic intellectual pursuits and collective well-being within academia.
Neoliberal labyrinth: Epistemic freedom and knowledge production in higher education in the Global South
Authors: Ndlovu Sibonokuhle , Woldegiorgis Emnet T.
Published: November 2024

In the 21st century, knowledge has become the driving force behind societal progress, emphasising the need for higher education to produce contextually relevant knowledge that addresses the multifaceted challenges faced by local communities. It is in this respect that knowledge needs to be generated through one’s position of epistemic location in higher education. However, academics positioned at the pinnacle of knowledge production in higher education find themselves entangled in a global crossroads. On the one hand, they are expected to exercise epistemic freedom by producing knowledge from their centralities and unique positionalities. On the other hand, they are constrained by the pervasive influence of neoliberalism, a paradigm that dictates that knowledge production should be subservient to market dynamics. To interrogate the intricacies of the impediments placed on academics seeking to exercise their epistemic freedom, this article utilises a desktop literature review, underscored by the theoretical framework of Decolonial Theory. The examination elucidates how these constraints hinder the production of knowledge from the centrality of the Global South. The article draws examples from South African higher education as a point of reference, providing examples that underscore the global challenges of neoliberal policies in higher education. Central to the thesis advanced in this article is the contention that, within the existing neoliberal framework of higher education driven by market forces and productivity imperatives, the production of knowledge from the position of one’s centrality is limited and the ability to produce locally relevant knowledge is fundamentally restricted. Consequently, the epistemic freedom of academics within higher education across the Global South is imperilled.Contribution: In response to these challenges, this article engages in an academic discourse on potential strategies for reclaiming epistemic freedom within the prevailing neoliberal milieu of higher education.
Study of Molecular Markers in Glioma and Their Association with Clinicopathological Features
Authors: Singh Alka , Singh Anurag , Jaiswal Awadhesh Kumar , Agrawal Sarita , Jaiswal Sushila
Published: November 2024

Context:

Central nervous system tumors are a major cause of morbidity and mortality worldwide. The most prevalent type of primary brain tumor is glioma. The exploration of significant genetic, epigenetic, and transcriptional abnormalities has not only improved our understanding of glioma pathogenesis but has also revealed that these molecular alterations can serve as useful diagnostic markers for more precise classification and are linked to better treatment response and prognosis. Hence, incorporating molecular markers into routine tumor classification is a major priority in modern glioma diagnostics.

Aim:

The aim is to assess the mutation status of isocitrate dehydrogenase (IDH)-1, alpha-thalassemia/mental retardation syndrome X-linked (ATRX), and tumor protein 53 in glioma, and look for their association with various clinicopathological features.

Methodology:

A single-center prospective cohort study, where all biopsies of glioma (January 2019 to July 2020) were evaluated, and immunohistochemistry was performed to assess the expression of IDH-1, ATRX, p53, and Ki-67 index. The data were analyzed using IBM SPSS-24 software.

Results:

Immunohistochemistry was performed in 123 consecutive cases of glioma. IDH-1 mutation was noted in 54 (43.9%) cases and these patients frequently presented with “seizures” (P = 0.006). The expression was maximum in World Health Organization (WHO) grade 2 tumors (65.4%) (P < 0.001), with the highest frequency in oligodendrogliomas (100% in WHO grade 2 and 3). Furthermore, these tumors showed lower proliferative indices (P = 0.001). ATRX mutation was noted in 59 (48%) and p53 overexpression was noted in 76 (61.8%) cases. These mutations were significantly associated with astrocytic phenotype (P = 0.03).

Conclusions:

Molecular characterization of glioma is an important step in modern glioma diagnostics and immunohistochemistry can play an important role. IDH-1 mutation is commonly observed in adults, frontal lobe location, patients presenting with seizures, and WHO grade 2 tumors with the highest frequencies in oligodendrogliomas. ATRX and p53 can be used as surrogate markers for tumors of astrocytic lineage.
The Varying Faces of MOGAD: A Case Series
Authors: Poovathingal Mary Anne
Published: November 2024

Myelin oligodendrocyte glycoprotein-immunoglobulin G (MOG-IgG)-associated disease (MOGAD) is an immune mediated demyelinating disorder initially described as a subtype of neuromyelitis optica spectrum disorder with antibodies against MOG. Recently, it has been described as a separate disease entity with unique clinical and radiological features. We herein report the clinical details of eight MOG-IgG-positive patients to highlight the peculiar and varied clinical and radiological features of this condition. Encephalitis was the most common presenting feature in our study, especially in pediatric cases. Tremors and parkinsonism were noted in four cases, of which one was a child. Monophasic course was noted in seven patients. Relapsing course was noted only in a single patient who was initiated on immunosuppressants. The clinical pictures highlight the importance of screening for anti-MOG in individuals with encephalitic and multiaxial symptoms in spite of having normal imaging. Acute demyelinating encephalomyelitis such as imaging features in the pediatric age group may also be a consideration for MOG testing.
Rapid Shallow Breathing Index and Ultrasonographic Diaphragmatic Parameters as Predictors of Weaning Outcome in Critically Ill Patients on Mechanical Ventilation
Authors: Swamy Akshay Hiryur Manjunatha , Kumararadhya Girish Bandigowdanahalli , Shivaramu Darshini , Challakere Gurudatt Lakshminarain , Chaitanya Krishna Ukkalam
Published: November 2024

Background:

Successful weaning is a crucial element in care toward critically ill patients on mechanical ventilation. An attempt was made to propose and assess a reliable predictor of weaning outcome.

Materials and Methods:

A prospective observational study was conducted on 76 patients on mechanical ventilation, assessed by Acute Physiology and Chronic Health Evaluation II (APACHE II) score. For all these patients we calculate Rapid shallow breathing index (RSBI), Ultrasonographic diaphragmatic parameters namely diaphragmatic excursion (DEx), diaphragmatic thickening fraction (DTF) and diaphragmatic contraction velocity (DCV). Values were compared among patients with two groups of successful and failed weaning outcomes, respectively, and statistically analyzed.

Results:

Of 76 patients included in the study, with ultrasonographic diaphragmatic parameters being measured 30 min into SBT, 71 patients tolerated spontaneous breathing test (SBT) for 2 h and were extubated. Of these, 61 patients did not require reintubation or any form of ventilatory support within 48 h after extubation. There was a statistically significant difference in APACHE II scores, duration of ventilation, oxygen saturation levels, RSBI, DEx, DTF, and DCV between groups of patients who showed successful and failed weaning from mechanical ventilation. There were a significant positive correlation between the duration of ventilation and the RSBI and a significant negative correlation between DEx, DCV, DTF, and duration of ventilation. As predictors of weaning outcome, RSBI showed the best validity, followed by DCV, DTF, and DEx.

Conclusion:

RSBI can be reliably used as a predictor of weaning outcome in critically ill patients on mechanical ventilation.
Advancing extracorporeal carbon dioxide removal technology: bridging basic science and clinical practice
Authors: Lassola Sergio , Battaglini Denise , De Rosa Silvia
Published: November 2024

Recently, advancements in extracorporeal carbon dioxide removal (ECCO2R) technology have markedly enhanced its clinical applicability and efficacy for managing severe respiratory conditions. This review highlights critical innovations in ECCO2R, such as advanced catheter technologies, active mixing methods, and biochemical enhancements, which have substantially improved gas exchange efficiency and broadened the scope of ECCO2R applications. Integrating ECCO2R into acute and chronic respiratory care has led to a shift toward more mobile and less invasive modalities, promising for extending ECCO2R usage from intensive care units to home settings. By examining these technological advancements and their clinical impacts, this paper outlines the potential future directions of ECCO2R technology, emphasizing its role in transforming respiratory care practices and enhancing patient outcomes.
Association between ethylene oxide exposure and osteoarthritis risk mediated by oxidative stress: evidence from NHANES 2013–2020
Authors: Yang Xinyue , Wang Jianwen , Wei Chengcheng , Tian Jia , Yan Lizhao , Huang Qishun
Published: November 2024

Ethylene oxide is extensively used for sterilizing medical equipment, and its carcinogenicity has been well documented. Furthermore, the onset of multiple diseases, including diabetes and hypertension, has been demonstrated to be associated with exposure to this compound. However, its association with osteoarthritis risk remains elusive. The study analyzed data from the National Health and Nutrition Examination Survey from 2013–2020, which included 6088 American adults, among whom 763 (12.5%) were diagnosed with osteoarthritis. We utilized a weighted generalized linear model to assess the correlation between ethylene oxide exposure levels and osteoarthritis risk. This study used mediation analysis to assess the functions of indicators of oxidative stress (γ-glutamyl transferase) and inflammation (alkaline phosphatase, white blood cell count, neutrophil count, and lymphocyte count) as mediators of how ethylene oxide affects osteoarthritis. The analysis revealed that elevated levels of ethylene oxide were correlated with a higher risk of osteoarthritis, even when controlling for other variables. The odds of developing osteoarthritis were 1.86 times higher in the fourth quartile than in the first quartile (95% confidence interval: 1.20–2.88, P = 0.0097, P for trend = 0.0087). Subgroup analyses indicated consistency across different cohorts. Mediation analysis revealed that oxidative stress (γ-glutamyl transferase), not inflammation, was the mediator linking ethylene oxide levels to the risk of osteoarthritis. This finding in a sample of American adults revealed a direct relationship between exposure to ethylene oxide and increased osteoarthritis risk. Oxidative stress has been suggested as a possible biological explanation for osteoarthritis caused by ethylene oxide.