bluuebunny/crossref_metadata_2025 huggingface dataset:
Multi‐channel seismic reflection study of tectonic–sedimentary features and subduction initiation in the middle Kyushu–Palau Ridge and adjacent basins
Authors: Ke Qin , Fanghui Hou , Qizhen Du , Kai Lu , Jingtao Zhao , Panfeng Li , Xiangjun Meng , Wei Huang , Gang Hu , Jun Sun , Xiaohan Gong
Published: December 2023

The Kyushu–Palau Ridge and adjacent basins are ideal locations for investigating the formation and evolution of marginal seas and initiation of plate subduction. In this study, the tectonic–sedimentary features and crustal structure of the Kyushu–Palau Ridge and adjacent basins were investigated using newly obtained deep seismic reflection and borehole data. The initial mechanism of subduction in the West Philippine Sea and its tectonic evolution are discussed. Two sets of sedimentary strata with different provenances occur in the eastern Philippine Basin. The thickness of the lower strata is variable, and most of the sediment was sourced from island arc volcanism on the Kyushu–Palau Ridge. These strata thicken towards the Kyushu–Palau Ridge, and volcaniclastic rock aprons are developed near the foot of the Kyushu–Palau Ridge. The upper strata have a relatively uniform thickness and comprise fine‐grained, deep‐water marine sediments. The crustal thickness of the West Philippine and Parece Vela Basins is 6–7 km, which is similar to the global average thickness of oceanic crust. The Moho beneath the West Philippine Basin has a broad and undulating form that mimics the thickness of the oceanic crust beneath the sediments. The depth to the Moho decreases towards the Central Basin Spreading Center. The Moho beneath the Parece Vela Basin is planar, which is in contrast to the undulating nature of the oceanic crust base in this area. The West Philippine Basin may have been located at the northern margin of Australia in the Southern Hemisphere during the Mesozoic. It developed gradually in a continental margin arc as a result of Palaeogene inter‐arc and oceanic extension and contains fragments of continental crust. Seismic profiles and drillhole data in the West Philippine Basin reveal that compression occurred during the Eocene. Subduction along the paleo‐Izu–Bonin–Mariana arc may have been caused by the far‐field effects of the India–Asia collision. Subduction was accompanied by lateral propagation and a compressive stress field until the break‐up of the island arc at ca. 30 Ma.
Fruit production in coffee (Coffea arabica L.) crops is enhanced by the behaviour of wild bees (Hymenoptera: Apidae)
Authors: Escobar‐González Denisse , Landaverde‐González Patricia , Casiá‐Ajché Quebin Bosbely , Morales‐Siná Javier , Cardona Edson , Mejía‐Coroy Alfredo , Enríquez Eunice
Published: December 2023

Changes in floral visitors' diversity and community composition have been reported to affect coffee production, which optimal growing conditions are cool to warm tropical climates found in the coffee belt. However, few studies have focused on understanding how insects' foraging behaviour (e.g., contact with floral reproductive organs) relates with coffee production. Thus, it is important to consider floral visitors' foraging behaviour, as this can influence the transfer of conspecific pollen required for plant fertilisation, the efficiency of floral visitors and improve the pollination service provided. Here, we assessed how foraging behaviour of honeybees and stingless bees affects coffee fruit set and fruit weight in conventional and agroecological managed crops. We quantified local floral resources and recorded diversity, abundance and behaviour of floral visitors at eight pairs of sites with agroecological and conventional management systems to assess how foraging behaviour of honeybees and stingless bees affects coffee fruit set and fruit weight in both types of managed crops. We found that the managed honeybee Apis mellifera and three wild bees Tetragonisca angustula, Scaptotrigona mexicana and Partamona bilineata are the principal floral visitors of coffee crops in Guatemala, whose total abundance but not richness was higher in agroecological areas. Regarding their behaviours, we observed that the average number of flowers visited by P. bilineata and its behaviour of touching the nectaries of coffee flowers were positively related to fruit set, while only the percentage of A. mellifera carrying pollen was positively related with fruit weight, suggesting that although A. mellifera is found in large quantities, wild bees are also efficient pollinators of coffee in the region. Our findings also suggest that in other tropical regions where coffee is grown and honeybees have been observed as a primary pollinator, wild bees may play an important role when considering their behaviour. In the same way, coffee farms in Guatemala are a representation of the diversity of agroecosystems found worldwide, and thus, the study of foraging behaviour of managed and wild bees and the conservation of wild bee species in different coffee agroecosystems should be emphasised to improve the production of coffee and other cash crops.
Environmental scanning, cross‐functional coordination and the adoption of green strategies: An information processing perspective
Authors: Song Yiru , Ali Adnan , Wang Gang , Wang Wenna
Published: December 2023

Information processing theory was applied to explore how external information processing and internal information sharing affect a firm's choice of a green strategy. Analyses of two datasets from Chinese manufacturing firms reveal that environment scanning and inter‐functional coordination are important drivers for the adoption of a green strategy. Specifically, firms that engage in extensive environment scanning tend to adopt reactive as opposed to proactive green strategies, whereas firms with effective inter‐functional coordination mechanisms tend to adopt proactive rather than reactive green strategies. An entrepreneurial orientation plays a complementary role in a firm's choice of green strategy. It increases the likelihood that a firm will adopt proactive rather than reactive green strategies, and it further strengthens the influences of environment scanning and inter‐functional coordination. Overall, these findings offer significant implications for firms regarding how and under what conditions they should adopt green strategies.
Low foraging rates drive large insectivorous bats away from urban areas
Authors: Stidsholt Laura , Scholz Carolin , Hermanns Uwe , Teige Tobias , Post Martin , Stapelfeldt Bianca , Reusch Christine , Voigt Christian C.
Published: December 2023

Urbanization has significant impacts on wildlife and ecosystems and acts as an environmental filter excluding certain species from local ecological communities. Specifically, it may be challenging for some animals to find enough food in urban environments to achieve a positive energy balance. Because urban environments favor small‐sized bats with low energy requirements, we hypothesized that common noctules (Nyctalus noctula) acquire food at a slower rate and rely less on conspecifics to find prey in urban than in rural environments due to a low food abundance and predictable distribution of insects in urban environments. To address this, we estimated prey sizes and measured prey capture rates, foraging efforts, and the presence of conspecifics during hunting of 22 common noctule bats equipped with sensor loggers in an urban and rural environment. Even though common noctule bats hunted similar‐sized prey in both environments, urban bats captured prey at a lower rate (mean: 2.4 vs. 6.3 prey attacks/min), and a lower total amount of prey (mean: 179 vs. 377 prey attacks/foraging bout) than conspecifics from rural environments. Consequently, the energy expended to capture prey was higher for common noctules in urban than in rural environments. In line with our prediction, urban bats relied less on group hunting, likely because group hunting was unnecessary in an environment where the spatial distribution of prey insects is predictable, for example, in parks or around floodlights. While acknowledging the limitations of a small sample size and low number of spatial replicates, our study suggests that scarce food resources may make urban habitats unfavorable for large bat species with higher energy requirements compared to smaller bat species. In conclusion, a lower food intake may displace larger species from urban areas making habitats with high insect biomass production key for protecting large bat species in urban environments.
In situ reaction synthesis of high‐entropy carbide/diboride composite with high mechanical properties
Authors: Tan Yongqiang , Liao Wei , Teng Zhen , Zhou Xiaosong
Published: December 2023

A novel technique to simultaneously lower the synthesis temperature of high‐entropy carbides and maintain their high mechanical properties was proposed. Certain amount of carbon vacancies was first introduced to significantly lower the temperature down to 2000°C for uniform elemental distributions in high‐entropy carbides. Those carbon vacancies were then fully eliminated through the reaction between high‐entropy carbides and certain amount of boron carbide. Concomitantly, the high‐entropy boride phase was formed. The elimination of carbon vacancies and the formation of high‐entropy boride phase significantly improved the mechanical properties of the high‐entropy carbides. A high mechanical strength over 500 MPa can be obtained by phase optimization.
Using a novel ambulatory monitoring system to support patient safety on an acute infectious disease ward during an unfolding pandemic
Authors: Buss Annika , Areia Carlos , Biggs Christopher , Edmundson Holly , Young Louise , Roman Cristian , Santos Mauro , Tarassenko Lionel , Watkinson Peter , Vollam Sarah
Published: December 2023

Aim

To gain staff feedback on the implementation and impact of a novel ambulatory monitoring system to support coronavirus patient management on an isolation ward.

Design

Qualitative service evaluation.

Methods

Semi‐structured interviews were conducted with 15 multidisciplinary isolation ward staff in the United Kingdom between July 2020 and May 2021. Interviews were audio‐recorded, transcribed and analysed using thematic analysis.

Findings

Adopting Innovation to Assist Patient Safety was identified as the overriding theme. Three interlinked sub‐themes represent facets of how the system supported patient safety. Patient Selection was developed throughout the pandemic, as clinical staff became more confident in choosing which patients would benefit most. Trust In the System described how nurses coped with discrepancies between the ambulatory system and ward observation machines. Finally, Resource Management examined how, once trust was built, staff perceived the ambulatory system assisted with caseload management. This supported efficient personal protective equipment resource use by reducing the number of isolation room entries. Despite these reported benefits, face‐to‐face contact was still highly valued, despite the risk of coronavirus exposure.

Conclusion

Hospital wards should consider using ambulatory monitoring systems to support caseload management and patient safety. Patients in isolation rooms or at high risk of deterioration may particularly benefit from this additional monitoring. However, these systems should be seen as an adjunct to nursing care, not a replacement.

Implications for the Profession and/or Patient Care

Nurses valued ambulatory monitoring as a means of ensuring the safety of patients at risk of deterioration and prioritizing their workload.

Impact

The findings of this research will be useful to all those developing or considering implementation of ambulatory monitoring systems in hospital wards.

Reporting Method

This manuscript follows the Consolidated criteria for Reporting Qualitative Research (COREQ) guidelines with inclusion of relevant SQUIRE guidelines for reporting quality improvement.

Patient or Public Contribution

No Patient or Public Contribution.
Severe asthma in horses is associated with increased airway innervation
Authors: Leduc Laurence , Leclère Mathilde , Gauthier Laurie Girardot , Marcil Olivier , Lavoie Jean‐Pierre
Published: December 2023

Background

Altered innervation structure and function contribute to airway hyperresponsiveness in human asthma, yet the role of innervation in airflow limitation in asthma in horses remains unknown.

Hypothesis

To characterize peribronchial innervation in horses with asthma. We hypothesized that airway innervation increases in horses with asthma compared with controls.

Animals

Formalin‐fixed lung samples from 8 horses with severe asthma and 8 healthy horses from the Equine Respiratory Tissue Biobank. Ante‐mortem lung function was recorded.

Methods

Blinded case‐control study. Immunohistochemistry was performed using rabbit anti‐s100 antibody as a neuronal marker for myelinating and non‐myelinating Schwann cells. The number and cumulative area of nerves in the peribronchial region and associated with airway smooth muscle were recorded using histomorphometry and corrected for airway size.

Results

Both the number (median [IQR]: 1.87 × 10−5 nerves/μm2 [1.28 × 10−5]) and the cumulative nerve area (CNA; 1.03 × 10−3 CNA/μm2 [1.57 × 10−3]) were higher in the peribronchial region of horses with asthma compared with controls (5.17 × 10−6 nerves/μm2 [3.76 × 10−6], 4.14 × 10−4 CNA/μm2 [2.54 × 10−4], Mann‐Whitney, P = .01). The number of nerves within or lining airway smooth muscle was significantly higher in horses with asthma (4.47 × 10−6 nerves/μm2 [5.75 × 10−6]) compared with controls (2.26 × 10−6 nerves/μm2 [1.16 × 10−6], Mann‐Whitney, P = .03).

Conclusions and Clinical Importance

Asthma in horses is associated with greater airway innervation, possibly contributing to airway smooth muscle remodeling and exacerbating severity of the disease.
Oncological impact of perioperative blood transfusion in bladder cancer patients undergoing radical cystectomy: Do we need to consider storage time of blood units, donor age, or gender matching?
Authors: Ebner Benedikt , Fleckenstein Franka , Volz Yannic , Eismann Lennert , Hermans Julian , Buchner Alexander , Enzinger Benazir , Weinhold Philipp , Wichmann Christian , Stief Christian Georg , Humpe Andreas , Pyrgidis Nikolaos , Schulz Gerald Bastian
Published: December 2023

Background

The oncological impact of perioperative blood transfusions (PBTs) of patients undergoing radical cystectomy (RC) because of bladder cancer (BCa) has been a controversial topic discussed in recent years. The main cause for the contradictory findings of existing studies might be the missing consideration of the storage time of red blood cell units (BUs), donor age, and gender matching.

Study Design and Methods

We retrospectively analyzed BCa patients who underwent RC in our department between 2004 and 2021. We excluded patients receiving BUs before RC, >10 BUs, or RC in a palliative setting. We assessed the effect of blood donor characteristics and storage time on overall survival (OS) and cancer‐specific survival (CSS) through univariate and multivariable Cox regression analysis. We also performed a propensity score matching with patients who received BUs and patients who did not on a 1:1 ratio.

Results

We screened 1692 patients and included 676 patients for the propensity score matching. In the multivariable analysis, PBT was independently associated with worse OS and CSS (p < .001). Postoperative transfusions were associated with better OS (p = .004) and CSS (p = .008) compared to intraoperative or mixed transfusions. However, there was no influence of blood donor age, storage time, or gender matching on prognosis.

Discussion

In our study of BCa patients undergoing RC, we demonstrate that PBT, especially if administered intraoperatively, is an independent risk factor for a worse prognosis.

However, storage time, donor age, or gender matching did not negatively affect oncological outcomes. Therefore, the specific selection of blood products does not promise any benefits.
Genomic analysis across 53 canine cancer types reveals novel mutations and high clinical actionability potential
Authors: Sakthikumar Sharadha , Warrier Manisha , Whitley Derick , Facista Salvatore , Adkins Jonathan , Aman Sara , Tsinajinnie Darwin , Duran Natalie , Siravegna Giulia , Ahmed Zeeshan , Day Kenneth , Jenkins Brooklyn , Patel Nidhi , Ryden Kirk , Nadai Joe , Banovich Kathryn , Powers Barbara , Edwards Jeffrey , Steinberg Jennifer , Fielder Susan , Wong Shukmei , Byron Sara A. , Izatt Tyler , Zismann Victoria , Boateng Martin , Zhu Zhanyang , Chuang Han‐Yu , Trent Jeffrey M. , Haworth David , Chon Esther , Hendricks William , Wang Guannan
Published: December 2023

A genomic understanding of the oncogenic processes and individual variability of human cancer has steadily fueled improvement in patient outcomes over the past 20 years. Mutations within tumour tissues are routinely assessed through clinical genomic diagnostic assays by academic and commercial laboratories to facilitate diagnosis, prognosis and effective treatment stratification. The application of genomics has unveiled a wealth of mutation‐based biomarkers in canine cancers, suggesting that the transformative principles that have revolutionized human cancer medicine can be brought to bear in veterinary oncology. To advance clinical genomics and genomics‐guided medicine in canine oncology, we have developed and validated a canine cancer next‐generation sequencing gene panel for the identification of multiple mutation types in clinical specimens. With this panel, we examined the genomic landscapes of 828 tumours from 813 dogs, spanning 53 cancer types. We identified 7856 alterations, encompassing copy number variants, single nucleotide variants, indels and internal tandem duplications. Additionally, we evaluated the clinical utility of these alterations by incorporating a biomarker framework from comprehensive curation of primary canine literature and inferences from human cancer genomic biomarker literature and clinical diagnostics. Remarkably, nearly 90% of the cases exhibited mutations with diagnostic, prognostic or therapeutic implications. Our work represents a thorough assessment of genomic landscapes in a large cohort of canine cancers, the first of its kind for its comprehensive inclusion of multiple mutation types and structured annotation of biomarkers, demonstrating the clinical potential of leveraging mutation‐based biomarkers in veterinary oncology.
A neuronal coping mechanism linking stress-induced anxiety to motivation for reward
Authors: Klenowski Paul M. , Zhao-Shea Rubing , Freels Timothy G. , Molas Susanna , Zinter Max , M’Angale Peter , Xiao Cong , Martinez-Núñez Leonora , Thomson Travis , Tapper Andrew R.
Published: December 2023

Stress coping involves innate and active motivational behaviors that reduce anxiety under stressful situations. However, the neuronal bases directly linking stress, anxiety, and motivation are largely unknown. Here, we show that acute stressors activate mouse GABAergic neurons in the interpeduncular nucleus (IPN). Stress-coping behavior including self-grooming and reward behavior including sucrose consumption inherently reduced IPN GABAergic neuron activity. Optogenetic silencing of IPN GABAergic neuron activation during acute stress episodes mimicked coping strategies and alleviated anxiety-like behavior. In a mouse model of stress-enhanced motivation for sucrose seeking, photoinhibition of IPN GABAergic neurons reduced stress-induced motivation for sucrose, whereas photoactivation of IPN GABAergic neurons or excitatory inputs from medial habenula potentiated sucrose seeking. Single-cell sequencing, fiber photometry, and optogenetic experiments revealed that stress-activated IPN GABAergic neurons that drive motivated sucrose seeking express somatostatin. Together, these data suggest that stress induces innate behaviors and motivates reward seeking to oppose IPN neuronal activation as an anxiolytic stress-coping mechanism.
Distinct microbial communities degrade cellulose diacetate bioplastics in the coastal ocean
Authors: Sun Yanchen , Mazzotta Michael G. , Miller Carolyn A. , Apprill Amy , Izallalen Mounir , Mazumder Sharmistha , Perri Steven T. , Edwards Brian , Reddy Christopher M. , Ward Collin P.
Published: December 2023

Cellulose diacetate (CDA) is a bio-based plastic widely used in consumer products. CDA is a promising alternative to conventional thermoplastics due to its susceptibility to biodegradation in various environments. Despite widespread evidence for the degradation of CDA, relatively little is known about the microorganisms that drive degradation, particularly in the ocean. Recently, we documented the biodegradation of CDA-based materials (i.e., fabric, film, and foam) in a continuous-flow natural seawater mesocosm on the timescales of months, as indicated by mass loss, enzyme activity, and respiration to carbon dioxide. These findings paved the way for the present study aimed at identifying key microbial taxa implicated in CDA degradation. Analysis based on 16S rRNA gene amplicon sequencing of bacteria and archaea revealed that material type, incubation time, material morphology (e.g., fabric vs film), and plasticizer content significantly influenced the microbial community structure. Differential abundance analysis revealed that bacterial taxa affiliated with the families of
Arenicellaceae
,
Cellvibrionaceae
,
Methyloligellaceae
,
Micavibrionaceae
,
Puniceicoccaceae
,
Spirosomaceae
, and
Thermoanaerobaculaceae
, and the order of
Pseudomonadales
potentially initiated the degradation (i.e., deacetylation) of CDA fabric and film. These taxa were notably distinct from CDA-degrading microbes reported in non-seawater environments. Collectively, the findings lend further support for CDA as a promising next-generation, high-utility, and low-environmental persistence bioplastic material.

IMPORTANCE

Cellulose diacetate (CDA) is a promising alternative to conventional plastics due to its versatility in manufacturing and low environmental persistence. Previously, our group demonstrated that CDA is susceptible to biodegradation in the ocean on timescales of months. In this study, we report the composition of microorganisms driving CDA degradation in the coastal ocean. We found that the coastal ocean harbors distinct bacterial taxa implicated in CDA degradation and these taxa have not been previously identified in prior CDA degradation studies, indicating an unexplored diversity of CDA-degrading bacteria in the ocean. Moreover, the shape of the plastic article (e.g., a fabric, film, or foam) and plasticizer in the plastic matrix selected for different microbial communities. Our findings pave the way for future studies to identify the specific species and enzymes that drive CDA degradation in the marine environment, ultimately yielding a more predictive understanding of CDA biodegradation across space and time.
Transcriptional expression of
PHR2
is positively controlled by the calcium signaling transcription factor Crz1 through its binding motif in the promoter
Authors: Jiang Linghuo , Xu Huihui , Wei Min , Gu Yiying , Yan Hongbo , Pan Lingxin , Wei Chunyu
Published: December 2023

GH72 family of β-(1,3)-glucanosyltransferases is required for cell wall biogenesis, morphogenesis, and virulence of fungi.
Candida albicans PHR2
is an acidic pH-induced gene that encodes a GH72 enzyme involved in cross-linking of β−1,3- and β−1,6-glucans in the cell wall. In response to environmental pH changes, the expression of
PHR2
is negatively regulated by the transcription factor CaRim101 through the pH-responsive Rim101 pathway. In this study, we have generated the inactivation mutant for
PHR2
through the CRISPR/Cas9 approach. The
PHR2
mutant is sensitive to both membrane- and cell wall-disturbing agents, SDS, Calcofluor white and casponfungi, but resistant to two azole antifungals, ketoconazole and fluconazole. Although the
PHR2
mutant is not sensitive to calcium stress, it becomes sensitive to high levels of calcium stress in the presence of cyclosporin A, the specific inhibitor of calcineurin. In addition, we have demonstrated that the expression of
PHR2
, the downstream target of CaRim101, is positively controlled by the transcription factor CaCrz1 through its binding motif (5′ A
-315
TAGGCACAG 3′) in the
PHR2
promoter. Furthermore, the deletion of
PHR2
increases the cell precipitation rate in cell culture under acidic conditions.

IMPORTANCE

The fungal cell wall consists of glucans, mannoproteins, and chitin and is essential for cell viability, morphogenesis, and pathogenesis. The enzymes of the GH72 family are responsible for ß-(1,3)-glucan elongation and branching, which is crucial for the formation of the glucan-chitin polymer at the bud neck of yeast cells. In the human fungal pathogen Candida albicans, there are five GH72 enzyme-encoding genes:
PHR1, PHR2, PHR3, PGA4
, and
PGA5
. It is known that expression of
PHR1
and
PHR2
is controlled by the pH-responsive Rim101 pathway through the transcription factor Rim101. In this study, we have demonstrated that the transcription expression of
PHR2
is also controlled by the transcription factor Crz1 through its binding motif in the promoter. Therefore, we have uncovered a dual-control mechanism by which
PHR2
expression is negatively regulated via CaRim101 through the pH-responsive pathway and positively modulated by CaCrz1 through the calcium/calcineurin signaling pathway.
Evaluation of the Xpert Xpress GBS test for rapid detection of group B
Streptococcus
in pregnant women
Authors: Wang Feiling , Yi Lehui , Ming Fang , Dong Rui , Wang Feng , Chen Ruirui , Hu Xiaoling , Chen Xuri , Sun Bo , Tang Yi-Wei , Zhu Yuanfang , Wu Lijuan
Published: December 2023

Group B
Streptococcus
(GBS) remains the leading cause of serious neonatal infection. Prenatal screening for vaginal and rectal GBS colonization is necessary in late pregnancy. Our objective was to compare the performance of four methods including the Xpert Xpress GBS, standard broth enrichment culture, real-time quantitative PCR (qPCR), and direct culture, for detecting of GBS in rectovaginal specimens. Among 939 pregnant women at 35–37 weeks of gestation recruited in the study, 168 (17.9%) were determined to be GBS-colonized by broth enrichment culture, the gold standard. The sensitivity and specificity of the Xpert Xpress GBS were 94.6% and 95.9%, respectively. The hands-on time and running time of the Xpert Xpress GBS were 1 minute and less than 1 hour, respectively. As a random-access, integrated device, the Xpert Xpress GBS test efficiently and accurately screens rectovaginal swabs for GBS colonization in pregnant women.

IMPORTANCE

This was the first study evaluating the performance of the Xpert Xpress group B
Streptococcus
(GBS) test using rectovaginal swabs from Chinese pregnant women. Compared to the other three assays, the Xpert Xpress GBS test demonstrated high sensitivity and specificity when screening 939 pregnant women for GBS in rectovaginal specimens. Additionally, its reduced time to obtain results makes it valuable for the rapid detection of GBS.
Vav proteins do not influence dengue virus replication but are associated with induction of phospho-ERK, IL-6, and viperin mRNA following DENV infection
in vitro
Authors: Cowell Evangeline , Jaber Hawraa , Kris Luke P. , Fitzgerald Madeleine G. , Sanders Valeria M. , Norbury Aidan J. , Eyre Nicholas S. , Carr Jillian M.
Published: December 2023

Vav proteins are guanine exchange factors that activate Rac1/RhoA signaling to regulate many biological processes including inflammatory responses. Here, the expression and functional significance of Vav’s were investigated during dengue virus (DENV) infection. In primary monocyte-derived macrophages, Vav1, −2, and −3 mRNA levels demonstrate variable responses to DENV infection and correlate with DENV-induced host inflammatory (IL-6 and TNF-α), antiviral (viperin), or cell adhesion [intercellular cell adhesion molecule (ICAM-1)] mRNA induction. Strong positive correlations were seen in particular for Vav2 with TNF-α and Vav3 with IL-6 mRNA. In the retinal pigmented epithelial cell line, ARPE-19, Vav2 was the main Vav expressed and was not affected by DENV infection. Heterologous Vav1 expression in ARPE-19 cells induced an increase in basal IL-6 mRNA but did not enhance DENV-induced mRNA responses. DENV RNA and DENV-induced viperin and IL-6 mRNA responses were also unaffected by Vav2 siRNA knockdown. Treatment of DENV-infected ARPE-19 cells with EHop-016 to block Vav signaling did not affect DENV RNA levels but increased DENV-mediated induction of IL-6 mRNA. More detailed assessment of DENV-induced responses to azathioprine, a clinically used immunosuppressant that can also block Vav signaling and act as a nucleoside analog, similarly demonstrated no change in DENV RNA levels but resulted in inhibition of DENV-induced phospho-ERK and increased DENV-induced-IL-6 and viperin mRNA in ARPE-19 cells. Thus, levels of Vav are associated with DENV-induced inflammatory responses, and blocking Vav signaling pathways does not compromise control of viral replication but may influence DENV-induced host responses.

IMPORTANCE

Dengue disease is characterized by an inflammatory-mediated immunopathology, with elevated levels of circulating factors including TNF-α and IL-6. If the damaging inflammatory pathways could be blocked without loss of antiviral responses or exacerbating viral replication, then this would be of potential therapeutic benefit. The study here has investigated the Vav guanine exchange factors as a potential alternative signaling pathway that may drive dengue virus (DENV)-induced inflammatory responses, with a focus on Vav1 and 2. While Vav proteins were positively associated with mRNA for inflammatory cytokines, blocking Vav signaling didn’t affect DENV replication but prevented DENV-induction of p-ERK and enhanced IL-6 (inflammatory) and viperin (antiviral) mRNA. These initial data suggest that Vav proteins could be a target that does not compromise control of viral replication and should be investigated further for broader impact on host inflammatory responses, in settings such as antibody-dependent enhancement of infection and in different cell types.
Val43 residue of NsrR is crucial for the nitric oxide response of
Salmonella
Typhimurium
Authors: Park Hee Jeong , Jeong Hye Won , Lee Choa , Lee Mi Rae , Choi Hojung , Kim Eungseok , Bang Iel Soo
Published: December 2023

In pathogenic bacteria, the flavohemoglobin Hmp is crucial in metabolizing the cytotoxic levels of nitric oxide (NO) produced in phagocytic cells, contributing to bacterial virulence. Hmp expression is predominantly regulated by the Rrf2 family transcription repressor NsrR in an NO-dependent manner; however, the underlying molecular mechanism in enterobacteria remains poorly understood. In this study, we identified Val43 of
Salmonella
Typhimurium NsrR (StNsrR) as a critical amino acid residue for regulating Hmp expression. The Val43-to-Ala-substituted mutant NsrR isolated through random and site-directed mutagenesis showed high binding affinity to the target DNA irrespective of NO exposure, resulting in a severe reduction in
hmp
transcription and slow NO metabolism in
Salmonella
under NO-producing conditions. Conversely, the Val43-to-Glu-substituted NsrR caused effects similar to
nsrR
null mutation, which directed
hmp
transcription and NO metabolism in a constitutive way. Comparative analysis of the primary sequences of NsrR and another NO-sensing Rrf2 family regulator, IscR, from diverse bacteria, revealed that Val43 of enterobacterial NsrR corresponds to Ala in
Pseudomonas aeruginosa
or
Streptomyces coelicolor
NsrR and Glu in enterobacterial IscR, all of which are located in the DNA recognition helix α3. The predicted structure of StNsrR in complex with the
hmp
DNA suggests dissimilar spatial stoichiometry in the interactions of Val43 and its substituted residues with the target DNA, consistent with the observed phenotypic changes in StNsrR Val43 mutants. Our findings highlight the discriminative roles of the NsrR recognition helix in regulating species-specific target gene expression, facilitating effective NO detoxification strategies in bacteria across diverse environments.

IMPORTANCE

The precise regulation of flavohemoglobin Hmp expression by NsrR is critical for bacterial fitness, as excessive Hmp expression in the absence of NO can disturb bacterial redox homeostasis. While the molecular structure of
Streptomyces coelicolor
NsrR has been recently identified, the specific molecular structures of NsrR proteins in enterobacteria remain unknown. Our discovery of the crucial role of Val43 in the DNA recognition helix α3 of
Salmonella
NsrR offers valuable insights into the Hmp modulation under NO stress. Furthermore, the observed amino acid polymorphisms in the α3 helices of NsrR proteins across different bacterial species suggest the diverse evolution of NsrR structure and gene regulation in response to varying levels of NO pressure within their ecological niches.
Impact of cord clamping on haemodynamic transition in term newborn infants
Authors: Chioma Roberto , Finn Daragh , Healy David B , Herlihy Ita , Livingstone Vicki , Panaviene Jurate , Dempsey Eugene M
Published: December 2023

Objective

To assess the haemodynamic consequences of cord clamping (CC) in healthy term infants.

Design

Cohort study.

Setting

Tertiary maternity hospital.

Patients

46 full-term vigorous infants born by caesarean section.

Interventions

Echocardiography was performed before CC, immediately after CC and at 5 min after birth.

Main outcome measures

Pulsed wave Doppler-derived cardiac output and the pulmonary artery acceleration time indexed to the right ventricle ejection time were obtained. As markers of loading fluctuations, the myocardial performance indexes and the velocities of the tricuspid and mitral valve annuli were determined with tissue Doppler imaging. Heart rate was derived from Doppler imaging throughout the assessments.

Results

Left ventricular output increased throughout the first minutes after birth (mean (SD) 222.4 (32.5) mL/kg/min before CC vs 239.7 (33.6) mL/kg/min at 5 min, p=0.01), while right ventricular output decreased (306.5 (48.2) mL/kg/min before vs 272.8 (55.5) mL/kg/min immediately after CC, p=0.001). The loading conditions of both ventricles were transiently impaired by CC, recovering at 5 min. Heart rate progressively decreased after birth, following a linear trend temporarily increased by CC. The variation in left ventricular output across the CC was directly correlated to the fluctuation of left ventricular preload over the same period (p=0.03).

Conclusions

This study illustrates the cardiovascular consequences of CC in term vigorous infants and offers insight into the haemodynamic transition from fetal to neonatal circulation in spontaneously breathing newborns. Strategies that aim to enhance left ventricular preload before CC may prevent complications of perinatal cardiovascular imbalance.
Treatment courses and outcomes of oesophageal atresia in patients with trisomy 18: a case series of 271 patients from a nationwide database in Japan
Authors: Kutsukake Mai , Konishi Takaaki , Fujiogi Michimasa , Takamoto Naohiro , Morita Kaori , Yasuhisa Ikuta , Hashimoto Yohei , Matsui Hiroki , Fushimi Kiyohide , Fujishiro Jun , Yasunaga Hideo
Published: December 2023

Objective

To describe characteristics, treatments and clinical outcomes of patients with trisomy 18 and oesophageal atresia, using a nationwide database in Japan.

Design

Descriptive study using a retrospective cohort.

Setting

A nationwide inpatient database including 90% of hospitals with neonatal intensive care units in Japan.

Patients

Patients hospitalised within a day after birth for both oesophageal atresia and trisomy 18 between July 2010 and March 2020.

Interventions

Radical surgery for oesophageal atresia.

Main outcome measures

Characteristics, treatment course and outcomes.

Results

Among 271 patients with both oesophageal atresia and trisomy 18, 70 patients underwent radical surgery for oesophageal atresia. Patients who underwent radical surgery were less likely to have severe cardiac anomalies (17% vs 32%; p=0.020), but more likely to undergo cardiac surgery (21% vs 9.5%; p=0.012) than those who did not. The overall in-hospital mortality was lower (54% vs 79%; p<0.001) and the median age at death was higher (210 days vs 39 days; p<0.001) in patients who underwent radical surgery than the others. Postoperative mortality within 30 days after radical surgery was 5.7%. Patients who underwent radical surgery were likely to be discharged to home (50% vs 18%; p<0.001), whereas the age at home discharge (median 314 days vs 216 days; p=0.19) and the requirement for each home treatment did not differ significantly by radical surgery.

Conclusion

This study provides information that will aid the clinical decision-making process for patients with oesophageal atresia and trisomy 18. Radical surgery may be a safe and feasible treatment option.
Marked increase in bone mineral density with oral phosphate and calcitriol in tumour-induced osteomalacia
Authors: Chakraborty Partha Pratim , Bhattacharjee Rana , Roy Ajitesh , Chowdhury Subhankar
Published: December 2023

Patients with osteomalacia have a low bone mineral density (BMD) and are often misdiagnosed as osteoporosis. A marked increase in BMD is noticed following successful treatment of osteomalacia. The biochemical hallmark of tumour-induced osteomalacia (TIO) is hypophosphatemia. Patients with TIO often have severe hypophosphatemic osteomalacia and dual-energy X-ray absorptiometry may demonstrate low BMD. Surgical removal of the phosphatonin-secreting lesion restores serum phosphate, corrects osteomalacia and is associated with a dramatic increase in BMD. We report two patients with TIO and low BMD, who were treated with oral phosphate and calcitriol supplementation. The percentage increase in BMD at 33 months was as high as 94.3% in areas with the lowest BMD at baseline. The BMD at 33 months was higher than the +2SD of the population-specific reference ranges, a finding not reported in surgically treated patients with TIO. An intermittent rise in parathyroid hormone following oral phosphate supplementation might have resulted in such findings.
Perioperative considerations for adult patients with Dravet syndrome in regional centres
Authors: Trinh Nguyen Hong Nhu Y , Reid Amy Susan , Robertson Barbara
Published: December 2023

Dravet syndrome (DS) is a rare and intractable severe form of epilepsy presenting in infancy with frequent prolonged myoclonic seizures and neurodevelopmental impairment, associated with aSCN1Agene mutation. Seizures are often triggered by temperature fluctuations and hyperthermia. This report presents a woman in her late adolescence with DS complicated with intractable catamenial epilepsy, a sex-specific form of epilepsy with seizure activity prominent during phases of the menstrual cycle. The patient underwent general anaesthesia for a hysteroscopy, cervical dilatation and endometrial curettage with Mirena insertion to improve seizure control. Her perioperative care was optimised for seizure prevention with continuation of antiepileptic medications, strict temperature monitoring and control, optimised anaesthetic agents encompassing induction with propofol and fentanyl with maintenance sevoflurane, followed by attentive postoperative care and monitoring. This case demonstrates that general anaesthesia can safely be delivered to adult patients with DS in rural and regional areas with thorough perioperative planning.
iPSC-derived natural killer cells expressing the FcγR fusion CD64/16A can be armed with antibodies for multitumor antigen targeting
Authors: Snyder Kristin M , Dixon Kate J , Davis Zachary , Hosking Martin , Hart Geoffrey , Khaw Melissa , Matson Anders , Bjordahl Ryan , Hancock Bryan , Shirinbak Soheila , Miller Jeffrey S , Valamehr Bahram , Wu Jianming , Walcheck Bruce
Published: December 2023

Background

Antibody therapies can direct natural killer (NK) cells to tumor cells, tumor-associated cells, and suppressive immune cells to mediate antibody-dependent cell-mediated cytotoxicity (ADCC). This antigen-specific effector function of human NK cells is mediated by the IgG Fc receptor CD16A (FcγRIIIA). Preclinical and clinical studies indicate that increasing the binding affinity and avidity of CD16A for antibodies improves the therapeutic potential of ADCC. CD64 (FcγRI), expressed by myeloid cells but not NK cells, is the only high affinity IgG Fc receptor and is uniquely capable of stably binding to free monomeric IgG as a physiological function. We have reported on the generation of the FcγR fusion CD64/16A, consisting of the extracellular region of CD64 and the transmembrane and cytoplasmic regions from CD16A, retaining its signaling and cellular activity. Here, we generated induced pluripotent stem cell (iPSC)-derived NK (iNK) cells expressing CD64/16A as a potential adoptive NK cell therapy for increased ADCC potency.

Methods

iPSCs were engineered to express CD64/16A as well as an interleukin (IL)-15/IL-15Rα fusion (IL-15RF) protein and differentiated into iNK cells. iNK cells and peripheral blood NK cells were expanded using irradiated K562-mbIL21-41BBL feeder cells and examined. NK cells, ovarian tumor cell lines, and therapeutic monoclonal antibodies were used to assess ADCC in vitro, performed by a DELFIA EuTDA assay or in real-time by IncuCyte assays, and in vivo. For the latter, we developed a xenograft mouse model with high circulating levels of human IgG for more physiological relevance.

Results

We demonstrate that (1) iNK-CD64/16A cells after expansion or thaw from cryopreservation can be coupled to therapeutic antibodies, creating armed iNK cells; (2) antibody-armed iNK-CD64/16A cells can be redirected by added antibodies to target new tumor antigens, highlighting additional potential of these cells; (3) cytokine-autonomous activity by iNK-CD64/16A cells engineered to express IL-15RF; and that (4) antibody-armed iNK-CD64/16A cells thawed from cryopreservation are capable of sustained and robust ADCC in vitro and in vivo, as determined by using a modified tumor xenograft model with high levels of competing human IgG.

Conclusions

iNK cells expressing CD64/16A provide an off-the-shelf multiantigen targeting platform to address tumor heterogeneity and mitigate antigen escape.
Allogeneic NK cells induce monocyte-to-dendritic cell conversion, control tumor growth, and trigger a pro-inflammatory shift in patient-derived cultures of primary and metastatic colorectal cancer
Authors: Toffoli Elisa C , van Vliet Amanda A , Verheul Henk W M , van der Vliet Hans J , Tuynman Jurriaan , Spanholtz Jan , de Gruijl Tanja D
Published: December 2023

Introduction

Natural killer (NK) cells are innate lymphocytes with a key role in the defense against tumors. Recently, allogeneic NK cell-based therapies have gained interest because of their ability to directly lyse tumor cells without inducing graft-versus-host disease. As NK cells are also able to influence the function of other immune cells (most notably dendritic cells (DC)), a better understanding of the effects of allogeneic NK cell products on the host immune system is required. In this study, we analyzed the effects of an allogeneic off-the-shelf NK cell product, on the tumor microenvironment (TME) of primary and metastatic colorectal cancer (pCRC and mCRC, respectively). Moreover, we explored if the combination of NK cells with R848, a toll-like receptors 7/8 ligand, could further enhance any pro-inflammatory effects.

Methods

Ex vivo expanded umbilical cord blood stem cell derived NK cells were co-cultured with pCRC or mCRC single-cell suspensions in the presence or absence of R848 for 5 days, during and after which flow cytometry and cytokine release profiling were performed.

Results

NK cells efficiently induced lysis of tumor cells in both pCRC and mCRC single-cell suspensions and thereby controlled growth rates during culture. They also induced differentiation of infiltrating monocytic cells to an activated DC phenotype. Importantly, this NK-mediated myeloid conversion was also apparent in cultures after tumor cell depletion and was further enhanced by combining NK cells with R848. Moreover, NK cells, and to a greater extent, the combination of NK cells and R848, triggered CD8+and CD4+T-cell activation as well as a reduction in activated regulatory T cell rates. Finally, the combination of NK cells and R848 induced a pro-inflammatory shift in the cytokine release profile resulting in higher levels of interferon (IFN)-γ, interleukin (IL)-2, IL-12p70, and IFN-α as well as a reduction in IL-6, in both pCRC and mCRC cultures.

Conclusion

Allogeneic NK cells engaged in favorable myeloid crosstalk, displayed effective antitumor activity and, when combined with R848, induced a pro-inflammatory shift of the CRC TME. These findings prompt the investigation of NK cells and R848 as a combination therapy for solid tumors.
Caspase-9 inhibition triggers Hsp90-based chemotherapy-mediated tumor intrinsic innate sensing and enhances antitumor immunity
Authors: Li Jingyang , Han Xiaoyu , Sun Mayu , Li Weida , Yang Guanghuan , Chen Huiyi , Guo Bao , Li Jingquan , Li Xiaoguang , Wang Hui
Published: December 2023

Background

Antineoplastic chemotherapies are dramatically efficient when they provoke immunogenic cell death (ICD), thus inducing an antitumor immune response and even tumor elimination. However, activated caspases, the hallmark of most cancer chemotherapeutic agents, render apoptosis immunologically silent. Whether they are dispensable for chemotherapy-induced cell death and the apoptotic clearance of cells in vivo is still elusive.

Methods

A rational cell-based anticancer drug library screening was performed to explore the immunogenic apoptosis pathway and therapeutic targets under apoptotic caspase inhibition. Based on this screening, the potential of caspase inhibition in enhancing chemotherapy-induced antitumor immunity and the mechanism of actions was investigated by various cells and mouse models.

Results

Heat shock protein 90 (Hsp90) inhibition activates caspases in tumor cells to produce abundant genomic and mitochondrial DNA fragments and results in cell apoptosis. Meanwhile, it hijacks Caspase-9 signaling to suppress intrinsic DNA sensing. Pharmacological blockade or genetic deletion of Caspase-9 causes tumor cells to secrete interferon (IFN)-β via tumor intrinsic mitochondrial DNA/the second messenger cyclic GMP–AMP (cGAS) /stimulator of interferon genes (STING) pathway without impairing Hsp90 inhibition-induced cell death. Importantly, both Caspase-9 and Hsp90 inhibition triggers an ICD, leading to the release of numerous damage-associated molecular patterns such as high-mobility group box protein 1, ATP and type I IFNs in vitro and remarkable antitumor effects in vivo. Moreover, the combination treatment also induces adaptive resistance by upregulating programmed death-ligand 1 (PD-L1). Additional PD-L1 blockade can further overcome this acquired immune resistance and achieve complete tumor regression.

Conclusions

Blockade of Caspase-9 signaling selectively provokes Hsp90-based chemotherapy-mediated tumor innate sensing, leading to CD8+T cell-dependent tumor control. Our findings implicate that pharmacological modulation of caspase pathway increases the tumor-intrinsic innate sensing and immunogenicity of chemotherapy-induced apoptosis, and synergizes with immunotherapy to overcome adaptive resistance.