bluuebunny/crossref_metadata_2025 huggingface dataset:
Fat-to-heart crosstalk in health and disease
Authors: Lodewijks Fleur , McKinsey Timothy A. , Robinson Emma L.
Published: March 2023

According to the latest World Health Organization statistics, cardiovascular disease (CVD) is one of the leading causes of death globally. Due to the rise in the prevalence of major risk factors, such as diabetes mellitus and obesity, the burden of CVD is expected to worsen in the decades to come. Whilst obesity is a major and consistent risk factor for CVD, the underlying pathological molecular communication between peripheral fat depots and the heart remains poorly understood. Adipose tissue (AT) is a major endocrine organ in the human body, with composite cells producing and secreting hormones, cytokines, and non-coding RNAs into the circulation to alter the phenotype of multiple organs, including the heart. Epicardial AT (EAT) is an AT deposit that is in direct contact with the myocardium and can therefore influence cardiac function through both mechanical and molecular means. Moreover, resident and recruited immune cells comprise an important adipose cell type, which can create a pro-inflammatory environment in the context of obesity, potentially contributing to systemic inflammation and cardiomyopathies. New mechanisms of fat-to-heart crosstalk, including those governed by non-coding RNAs and extracellular vesicles, are being investigated to deepen the understanding of this highly common risk factor. In this review, molecular crosstalk between AT and the heart will be discussed, with a focus on endocrine and paracrine signaling, immune cells, inflammatory cytokines, and inter-organ communication through non-coding RNAs.
Non-viral TRAC-knocked-in CD19KICAR-T and gp350KICAR-T cells tested against Burkitt lymphomas with type 1 or 2 EBV infection: In vivo cellular dynamics and potency
Authors: Braun Tobias , Pruene Alina , Darguzyte Milita , vom Stein Alexander F. , Nguyen Phuong-Hien , Wagner Dimitrios L. , Kath Jonas , Roig-Merino Alicia , Heuser Michael , Riehm Lucas L. , Schneider Andreas , Awerkiew Sabine , Talbot Steven R. , Bleich André , Figueiredo Constanca , Bornhäuser Martin , Stripecke Renata
Published: March 2023

Introduction

The ubiquitous Epstein–Barr virus (EBV) is an oncogenic herpes virus associated with several human malignancies. EBV is an immune-evasive pathogen that promotes CD8+ T cell exhaustion and dysregulates CD4+ T cell functions. Burkitt lymphoma (BL) is frequently associated with EBV infections. Since BL relapses after conventional therapies are difficult to treat, we evaluated prospective off-the-shelf edited CAR-T cell therapies targeting CD19 or the EBV gp350 cell surface antigen.

Methods

We used CRISPR/Cas9 gene editing methods to knock in (KI) the CD19CAR.CD28z or gp350CAR.CD28z into the T cell receptor (TCR) alpha chain (TRAC) locus.

Results

Applying upscaled methods with the ExPERT ATx® MaxCyte system, KI efficacy was ~20% of the total ~2 × 108 TCR-knocked-out (KO) generated cells. KOTCRKICAR-T cells were co-cultured in vitro with the gp350+CD19+ BL cell lines Daudi (infected with type 1 EBV) or with Jiyoye (harboring a lytic type 2 EBV). Both types of CAR-T cells showed cytotoxic effects against the BL lines in vitro. CD8+ KICAR-T cells showed higher persistency than CD4+ KICAR-T cells after in vitro co-culture with BL and upregulation of the activation/exhaustion markers PD-1, LAG-3, and TIM-3. Two preclinical in vivo xenograft models were set up with Nod.Rag.Gamma mice injected intravenously (i.v.) with 2 × 105 Daudi/fLuc-GFP or with Jiyoye/fLuc-GFP cells. Compared with the non-treated controls, mice challenged with BL and treated with CD19KICAR-T cells showed delayed lymphoma dissemination with lower EBV DNA load. Notably, for the Jiyoye/fLuc-GFP model, almost exclusively CD4+ CD19KICAR-T cells were detectable at the endpoint analyses in the bone marrow, with increased frequencies of regulatory T cells (Tregs) and TIM-3+CD4+ T cells. Administration of gp350KICAR-T cells to mice after Jiyoye/GFP-fLuc challenge did not inhibit BL growth in vivo but reduced the EBV DNA load in the bone marrow and promoted gp350 antigen escape. CD8+PD-1+LAG-3+ gp350KICAR-T cells were predominant in the bone marrow.

Discussion

The two types of KOTCRKICAR-T cells showed different therapeutic effects and in vivo dynamics. These findings reflect the complexities of the immune escape mechanisms of EBV, which may interfere with the CAR-T cell property and potency and should be taken into account for future clinical translation.
Advanced immunophenotyping: A powerful tool for immune profiling, drug screening, and a personalized treatment approach
Authors: Preglej Teresa , Brinkmann Marie , Steiner Günter , Aletaha Daniel , Göschl Lisa , Bonelli Michael
Published: March 2023

Various autoimmune diseases are characterized by distinct cell subset distributions and activation profiles of peripheral blood mononuclear cells (PBMCs). PBMCs can therefore serve as an ideal biomarker material, which is easily accessible and allows for screening of multiple cell types. A detailed understanding of the immune landscape is critical for the diagnosis of patients with autoimmune diseases, as well as for a personalized treatment approach. In our study, we investigate the potential of multi-parameter spectral flow cytometry for the identification of patients suffering from autoimmune diseases and its power as an evaluation tool for in vitro drug screening approaches (advanced immunophenotyping). We designed a combination of two 22-color immunophenotyping panels for profiling cell subset distribution and cell activation. Downstream bioinformatics analyses included percentages of individual cell populations and median fluorescent intensity of defined markers which were then visualized as heatmaps and in dimensionality reduction approaches. In vitro testing of epigenetic immunomodulatory drugs revealed an altered activation status upon treatment, which supports the use of spectral flow cytometry as a high-throughput drug screening tool. Advanced immunophenotyping might support the exploration of novel therapeutic drugs and contribute to future personalized treatment approaches in autoimmune diseases and beyond.
Tumor intrinsic and extrinsic functions of CD73 and the adenosine pathway in lung cancer
Authors: Kowash Ryan R. , Akbay Esra A.
Published: March 2023

The adenosine pathway is an exciting new target in the field of cancer immunotherapy. CD73 is the main producer of extracellular adenosine. Non-small cell lung cancer (NSCLC) has one of the highest CD73 expression signatures among all cancer types and the presence of common oncogenic drivers of NSCLC, such as mutant epidermal growth factor receptor (EGFR) and KRAS, correlate with increased CD73 expression. Current immune checkpoint blockade (ICB) therapies only benefit a subset of patients, and it has proved challenging to understand which patients might respond even with the current understanding of predictive biomarkers. The adenosine pathway is well known to disrupt cytotoxic function of T cells, which is currently the main target of most clinical agents. Data thus far suggests that combining ICB therapies already in the clinic with adenosine pathway inhibitors provides promise for the treatment of lung cancer. However, antigen loss or lack of good antigens limits efficacy of ICB; simultaneous activation of other cytotoxic immune cells such as natural killer (NK) cells can be explored in these tumors. Clinical trials harnessing both T and NK cell activating treatments are still in their early stages with results expected in the coming years. In this review we provide an overview of new literature on the adenosine pathway and specifically CD73. CD73 is thought of mainly for its role as an immune modulator, however recent studies have demonstrated the tumor cell intrinsic properties of CD73 are potentially as important as its role in immune suppression. We also highlight the current understanding of this pathway in lung cancer, outline ongoing studies examining therapies in combination with adenosine pathway targeting, and discuss future prospects.
A computer vision approach for analyzing label free leukocyte trafficking dynamics on a microvascular mimetic
Authors: Ahmad S. Danial , Cetin Mujdat , Waugh Richard E. , McGrath James L.
Published: March 2023

High-content imaging techniques in conjunction within vitromicrophysiological systems (MPS) allow for novel explorations of physiological phenomena with a high degree of translational relevance due to the usage of human cell lines. MPS featuring ultrathin and nanoporous silicon nitride membranes (µSiM) have been utilized in the past to facilitate high magnification phase contrast microscopy recordings of leukocyte trafficking events in a living mimetic of the human vascular microenvironment. Notably, the imaging plane can be set directly at the endothelial interface in a µSiM device, resulting in a high-resolution capture of an endothelial cell (EC) and leukocyte coculture reacting to different stimulatory conditions. The abundance of data generated from recording observations at this interface can be used to elucidate disease mechanisms related to vascular barrier dysfunction, such as sepsis. The appearance of leukocytes in these recordings is dynamic, changing in character, location and time. Consequently, conventional image processing techniques are incapable of extracting the spatiotemporal profiles and bulk statistics of numerous leukocytes responding to a disease state, necessitating labor-intensive manual processing, a significant limitation of this approach. Here we describe a machine learning pipeline that uses a semantic segmentation algorithm and classification script that, in combination, is capable of automated and label-free leukocyte trafficking analysis in a coculture mimetic. The developed computational toolset has demonstrable parity with manually tabulated datasets when characterizing leukocyte spatiotemporal behavior, is computationally efficient and capable of managing large imaging datasets in a semi-automated manner.
Combining preclinical tools and models to unravel tumor complexity: Jump into the next dimension
Authors: Miserocchi Giacomo , Bocchini Martine , Cortesi Michela , Arienti Chiara , De Vita Alessandro , Liverani Chiara , Mercatali Laura , Bravaccini Sara , Ulivi Paola , Zanoni Michele
Published: March 2023

Tumors are complex and heterogeneous diseases characterized by an intricate milieu and dynamically in connection with surrounding and distant tissues. In the last decades, great efforts have been made to develop novel preclinical models able to recapitulate the original features of tumors. However, the development of an in vitro functional and realistic tumor organ is still utopic and represents one of the major challenges to reproduce the architecture of the tumor ecosystem. A strategy to decrypt the whole picture and predict its behavior could be started from the validation of simplified biomimetic systems and then proceed with their integration. Variables such as the cellular and acellular composition of tumor microenvironment (TME) and its spatio-temporal distribution have to be considered in order to respect the dynamic evolution of the oncologic disease. In this perspective, we aim to explore the currently available strategies to improve and integrate in vitro and in vivo models, such as three-dimensional (3D) cultures, organoids, and zebrafish, in order to better understand the disease biology and improve the therapeutic approaches.
Atlantic cod (Gadus morhua) assessment approaches in the North and Baltic Sea: A comparison of environmental DNA analysis versus bottom trawl sampling
Authors: Kasmi Yassine , Blancke Tina , Eschbach Erik , Möckel Benita , Casas Laura , Bernreuther Matthias , Nogueira Pedro , Delfs Gertrud , Kadhim Samira , Meißner Timo , Rödiger Marcellus , Eladdadi Amina , Stransky Christoph , Hanel Reinhold
Published: March 2023

The assessment of fish stocks is often dependent on scientific trawl fisheries surveys, which are both invasive and costly. The analysis of environmental DNA (eDNA) from water samples is regarded as a non-invasive and cost-effective alternative, but meaningful performance evaluations are required for a wider application. The goal of this study was to comparatively analyze a newly developed, more sensitive real-time PCR based eDNA approach with bottom trawl fisheries catches to locally detect and quantify Atlantic cod (Gadus morhua) in the North and Baltic Seas. With a species-specificity of the qPCR assay of 100%, a minimal limit of 15 Cytochrome b eDNA copies was determined for the detection of cod. In addition, a Gaussian processing regression proved a significant correlation (95%) between eDNA (copies per L of water) and cod biomass (CPUE/Ha) found by bottom trawling. The results presented here prove the potential of eDNA analyses for quantitative assessments of commercial fish stocks in the open ocean, although additional comparative analyses are needed to demonstrate its performance under different oceanographic conditions.
The development of ocean governance for marine environment protection: Current legal system in Taiwan
Authors: Shih Yi-Che , Chen Wei Chung , Chen Tai-An Peter , Chang Chih-wei
Published: March 2023

Taiwan establishes Ocean Affairs Council (OAC) in 2018. Ocean governance has reached a new milestone. In 2019, the Ocean Basic Act was enacted. In 2020, the National Ocean Policy White Paper was published, meaning that Taiwan has specialized ocean authorities, regulations, enforcement units, and relevant mechanisms and policies. The Ocean Conservation Administration (OCA) is also responsible for marine environmental protection and conservation. To ensure good ocean governance, maintain marine resources, and protect the environment, the OCA has recently drafted the Marine Conservation Act for sustainable development. This article mainly reviews, analyzes, and compares Taiwan’s current marine-related laws and regulations and refers to the laws, policies, and mechanisms of other countries to provide suggestions on marine governance and the ongoing draft of the Marine Conservation Act.
Real-time detection of deep-sea hydrothermal plume based on machine vision and deep learning
Authors: Wang Xun , Cao Yanpeng , Wu Shijun , Yang Canjun
Published: March 2023

Recent years have witnessed an increase in applications of artificial intelligence (AI) in the detection of oceanic features with the tremendous success of deep learning. Given the unique biological ecosystems and mineral-rich deposits, the exploration of hydrothermal fields is both scientifically and commercially important. To achieve autonomous and intelligent sampling of the hydrothermal plume by using AUV, this paper proposes an innovative method for real-time plume detection based on the YOLOv5n deep learning algorithm designed with a light-weight neural network architecture to meet the requirements of embedded platforms. Ground truth labeler app LabelImg was used to generate the ground truth data from the plume dataset created by ourselves. To accurately and efficiently detect hydrothermal plumes using an embedded system, we improved the original structure of YOLOv5n in two aspects. First, SiLU activation functions in the model were replaced by ReLU activations at shallow layers and Hard-SiLU activations at deep layers to reduce the number of calculations. Second, an attention module termed Coordinate Attention (CA) was integrated into the model to improve its sensitivity to both channel and spatial features. In addition, a transfer learning training method was adopted to further improve the model’s accuracy and generalizability. Finally, we successfully deployed the proposed model in a low-cost embedded device (NVIDIA Jetson TX2 NX) by using the TensorRT inference engine. We then installed the Jetson TX2 NX into a hovering-type AUV as its vision processing unit and conducted a plume detection test in the water tank. The water tank experimental results demonstrated that the proposed method can achieve real-time onboard hydrothermal plume detection.
Convolution, aggregation and attention based deep neural networks for accelerating simulations in mechanics
Authors: Deshpande Saurabh , Sosa Raúl I. , Bordas Stéphane P. A. , Lengiewicz Jakub
Published: March 2023

Deep learning surrogate models are being increasingly used in accelerating scientific simulations as a replacement for costly conventional numerical techniques. However, their use remains a significant challenge when dealing with real-world complex examples. In this work, we demonstrate three types of neural network architectures for efficient learning of highly non-linear deformations of solid bodies. The first two architectures are based on the recently proposed CNN U-NET and MAgNET (graph U-NET) frameworks which have shown promising performance for learning on mesh-based data. The third architecture is Perceiver IO, a very recent architecture that belongs to the family of attention-based neural networks–a class that has revolutionised diverse engineering fields and is still unexplored in computational mechanics. We study and compare the performance of all three networks on two benchmark examples, and show their capabilities to accurately predict the non-linear mechanical responses of soft bodies.
Prepackaged formula low-residue diet vs. self-prepared low-residue diet before colonoscopy: A multicenter randomized controlled trial
Authors: Pan Peng , Gu Lun , Zhao Shengbing , Wang Shuling , Ma Jiayi , Fu Hongyu , Chen Youxiang , He Shuixiang , Tian Zibin , Xu Le , Feng Zhijie , Li Yanqing , Yang Zhuo , Yang Lei , Wang Wen , Hou Qian , Liu Ting , Li Chujun , Tian Dean , Wang Xiaodi , Gao Yongmei , Shi Hanping , Bai Yu , Li Zhaoshen
Published: March 2023

Background and aims

Compared with self-prepared LRD, a prepackaged low-residue diet (LRD) can improve patient compliance, but whether it can further improve the quality of bowel preparation is uncertain. The study aimed to compare the application of the prepackaged formula LRD with self-prepared LRD in bowel preparation for colonoscopy.

Methods

A multicenter randomized controlled trial was conducted in 15 centers. The eligible subjects were randomly assigned to one of two groups: the formula LRD group and the self-prepared LRD group. On the day before the colonoscopy, subjects in the self-prepared LRD group were instructed to consume a restricted LRD prepared by themselves, while subjects in the formula LRD group were given six bags of prepackaged formula LRD and instructed to consume them according to their individual need. The primary outcome was an adequate bowel preparation rate. Secondary outcomes mainly included Boston Bowel Preparation Scale (BBPS) scores, dietary restriction compliance rate, tolerance, satisfaction, adenoma detection rate (ADR), and adverse reactions. The trial was registered at ClinicalTrials.gov under the identifier NCT03943758.

Results

A total of 550 subjects were recruited. Compared with the self-prepared LRD group, the formula LRD group showed a higher adequate bowel preparation rate (94.5 vs. 80.4%; P < 0.01), BBPS scores (7.87 ± 1.13 vs. 6.75 ± 1.47; P < 0.01), dietary compliance rate (92.4 vs. 78.9%; P < 0.01), tolerance (P < 0.01 in degree of hunger, intensity of physical strength, and negative influence on daily activities), satisfaction (8.56 ± 1.61 vs. 7.20 ± 2.02; P < 0.01), and ADR (25.6 vs. 16.0%; P < 0.01). There was no significant difference in adverse reactions.

Conclusion

Compared with self-prepared LRD, the formula LRD showed similar safety and higher bowel preparation quality, compliance, and tolerance in bowel preparation. More formula LRDs could be designed according to different dietary habits and ethnic populations, and further researches are warranted to confirm their effect.

Clinical trial registration

https://register.clinicaltrials.gov, identifier: NCT03943758.
Disease modeling of ADAMTS9-related nephropathy using kidney organoids reveals its roles in tubular cells and podocytes
Authors: Yu Seyoung , Choi Yo Jun , Rim John Hoon , Kim Hye-Youn , Bekheirnia Nasim , Swartz Sarah Jane , Dai Hongzheng , Gu Shen Linda , Lee Soyeon , Nishinakamura Ryuichi , Hildebrandt Friedhelm , Bekheirnia Mir Reza , Gee Heon Yung
Published: March 2023

Introduction

Mutations in ADAMTS9 cause nephronophthisis-related ciliopathies (NPHP-RC), which are characterized by multiple developmental defects and kidney diseases. Patients with NPHP-RC usually have normal glomeruli and negligible or no proteinuria. Herein, we identified novel compound-heterozygous ADAMTS9 variants in two siblings with NPHP-RC who had glomerular manifestations, including proteinuria.

Methods

To investigate whether ADAMTS9 dysfunction causes NPHP and glomerulopathy, we differentiated ADAMTS9 knockout human induced pluripotent stem cells (hiPSCs) into kidney organoids. Single-cell RNA sequencing was utilized to elucidate the gene expression profiles from the ADAMTS9 knockout kidney organoids.

Results

ADAMTS9 knockout had no effect on nephron differentiation; however, it reduced the number of primary cilia, thereby recapitulating renal ciliopathy. Single-cell transcriptomics revealed that podocyte clusters express the highest levels of ADAMTS9, followed by the proximal tubules. Loss of ADAMTS9 increased the activity of multiple signaling pathways, including the Wnt/PCP signaling pathway, in podocyte clusters.

Conclusions

Mutations in ADMATS9 cause a glomerulotubular nephropathy in kidney and our study provides insights into the functional roles of ADMATS9 in glomeruli and tubules.
Determining a common understanding of interprofessional competencies for pre-registration health professionals in Aotearoa New Zealand: A Delphi study
Authors: Andersen Patrea , Broman Patrick , Tokolahi Ema , Yap Jia Rong , Brownie Sharon
Published: March 2023

There is growing awareness that factors such as the growing incidence of co-morbidity and increasing complexity of patient health needs cannot be addressed by health professionals practicing in isolation. Given this, there is an increasing emphasis on preparing students in health-related programs for effective interprofessional practice. Less clear, however, are the specific skills and clinical or learning opportunities necessary for students to develop effectiveness in interprofessional practice. These factors drove a team associated with a tertiary health education provider in Hamilton, New Zealand to transform traditional clinical student experiences in the form of an interprofessional student-assisted clinic. The clinic was intended, in part, to provide students with opportunities to learn and experience interprofessionalism in practice but was hampered by limited information available regarding the specific skill requirements necessary for students in New Zealand to learn in this context. In this Delphi study, we synthesize national expert opinion on student competency indicators necessary for effective interprofessional practice. The resultant set of indicators is presented and opportunities for application and further research discussed. The paper offers guidance to others seeking to innovate health curricula, develop novel service-oriented learning experiences for students, and foster interprofessional practice competence in the future health workforce.
Recent advances in the treatment of systemic sclerosis associated interstitial lung disease
Authors: Kamenova Antoniya , Tzouvelekis Argyris , Margaritopoulos George A.
Published: March 2023

Connective tissue diseases (CTDs) are a heterogenous group of systemic inflammatory disorders. The development of connective tissue disease-associated interstitial lung disease (CTD-ILD) is a key complication associated with significant morbidity and mortality. The aim of this review is to explore the pathogenesis of CTD-ILD and summarize the recent evidence from clinical trials for novel treatment options, including the role of antifibrotics and immunomodulatory therapies with a focus on systemic sclerosis associated ILD. Further clinical trials are ongoing to explore combination therapies and more targeted therapeutic options. Clinicians remain faced with the difficult challenge of appropriately selecting patients who will benefit from the available therapies and timing the start of therapy at the most suitable part of the disease course.
Long-term simulated microgravity alters gut microbiota and metabolome in mice
Authors: Yuan Lu , Zhang Rong , Li Xinlou , Gao Caiyun , Hu Xiangnan , Hussain Safdar , Zhang Linlin , Wang Moye , Ma Xiaoyu , Pan Qiuxia , Lou Xiaotong , Si Shaoyan
Published: March 2023

Spaceflight and microgravity has a significant impact on the immune, central nervous, bone, and muscle support and cardiovascular systems. However, limited studies are available on the adverse effects of long-term microgravity on the intestinal microbiota, metabolism, and its relationships. In this study, a ground-based simulated microgravity (SMG) mouse model was established to evaluate the impact of long-term microgravity on gut microbiota and metabolome. After 8 weeks of SMG, alterations of the intestinal microbiota and metabolites were detected using 16S rRNA sequencing and untargeted metabolomics. Compared to the control, no significant differences in α-diversity were observed at weeks 2, 4 and 8. Nevertheless, there were clear differences in community structures at different time points. The phylum Verrucomicrobia significantly declined from 2 to 8 weeks of SMG, yet the relative abundance of Actinobacteria and Deferribacteres expanded remarkably at weeks 8. SMG decreased the genus of Allobaculum and increased Bacteroides significantly throughout the period of 8 weeks. Besides, Genus Akkermansia, Gracilibacter, Prevotella, Odoribacter, Rothia, Sporosarcina, Gracilibacter, Clostridium, and Mucispirillum were identified as biomarkers for SMG group. Desulfovibrio_c21_c20, Akkermansia_muciniphila, and Ruminococcus_gnavus dropped at week 2, which tend to recover at week 4, except for Akkermansia_muciniphila. Bacteroides_uniformis and Faecalibacterium_prausnitzii declined significantly, while Ruminococcus_flavefaciens and Mucispirillum_schaedleri elevated at week 8. Furthermore, intestinal metabolome analysis showed that 129 were upregulated and 146 metabolites were downregulated in SMG. Long-term SMG most affected steroid hormone biosynthesis, tryptophan, cysteine, methionine, arginine, proline metabolism, and histidine metabolism. Correlated analysis suggested that the potential beneficial taxa Allobaculum, Akkermansia, and Faecalibacterium were negatively associated with tryptophan, histidine, arginine, and proline metabolism, but positively with steroid hormone biosynthesis. Yet Bacteroides, Lachnospiraceae_Clostridium, Rothia, Bilophila, and Coprococcus were positively correlated with arginine, proline, tryptophan, and histidine metabolism, while negatively associated with steroid hormone biosynthesis. These results suggest that Long-term SMG altered the community of intestinal microbiota, and then further disturbed intestinal metabolites and metabolic pathways, which have great potential to help understand and provide clues for revealing the mechanisms of long-term SMG involved diseases.
Characterization of a cyanobacterial rep protein with broad-host range and its utilization for expression vectors
Authors: Sakamaki Yutaka , Maeda Kaisei , Nimura-Matsune Kaori , Chibazakura Taku , Watanabe Satoru
Published: March 2023

Owing to their photosynthetic capabilities, cyanobacteria are regarded as ecologically friendly hosts for production of biomaterials. However, compared to other bacteria, tools for genetic engineering, especially expression vector systems, are limited. In this study, we characterized a Rep protein, exhibiting replication activity in multiple cyanobacteria and established an expression vector using this protein. Our comprehensive screening using a genomic library of Synechocystis sp. PCC 6803 revealed that a certain region encoding a Rep-related protein (here named Cyanobacterial Rep protein A2: CyRepA2) exhibits high autonomous replication activity in a heterologous host cyanobacterium, Synechococcus elongatus PCC 7942. A reporter assay using GFP showed that the expression vector pYS carrying CyRepA2 can be maintained in not only S. 6803 and S. 7942, but also Synechococcus sp. PCC 7002 and Anabaena sp. PCC 7120. In S. 7942, GFP expression in the pYS-based system was tightly regulated by IPTG, achieving 10-fold higher levels than in the chromosome-based system. Furthermore, pYS could be used together with the conventional vector pEX, which was constructed from an endogenous plasmid in S. 7942. The combination of pYS with other vectors is useful for genetic engineering, such as modifying metabolic pathways, and is expected to improve the performance of cyanobacteria as bioproduction chassis.
Immune profiling of experimental murine mastitis reveals conserved response to mammary pathogenic Escherichia coli, Mycoplasma bovis, and Streptococcus uberis
Authors: Schneider Peleg , Salamon Hagit , Weizmann Nathalie , Nissim-Eliraz Einat , Lysnyansky Inna , Shpigel Nahum Y.
Published: March 2023

Mastitis is one of the most prevalent and economically important diseases of dairy animals. The disease is caused by ascending bacterial infection through the teat canal. Among the most common mastitis-causing bacteria are Gram-negative coliforms, Gram-positive streptococci and staphylococci, and mycoplasma. The most prominent cellular hallmark of acute mammary infection is a massive recruitment of blood neutrophils into the tubular and alveolar milk spaces. The complex biological processes of leukocyte recruitment, activation, adhesion, and migration in the mammary gland remain largely elusive to date. While field research of mastitis in dairy animals contributed a lot to the development of mitigation, control, and even eradication programs, little progress was made toward understanding the molecular mechanisms underlying the pathogenesis of the disease. We report here experimental mastitis model systems in lactating mice challenged with field strains of common udder pathogens in dairy cows. We used these model systems to apply recently developed multiplex gene expression technology (Nanostring nCounter), which enabled us to study the expression of over 700 immune genes. Our analysis revealed a core of 100 genes that are similarly regulated and functionally or physically interacting in E. coli, M. bovis, and Strep uberis murine mastitis. Common significantly enriched gene sets include TNFɑ signaling via NFkB, Interferon gamma and alpha response, and IL6-JAK-STAT3 signaling. In addition, we show a significantly enriched expression of genes associated with neutrophil extracellular traps (NET) in glands challenged by the three pathogens. Ligand-receptor analysis revealed interactions shared by the three pathogens, including the interaction of the cytokines IL1β, IL1ɑ, and TNFɑ with their receptors, and proteins involved in immune cell recruitment such as complement C3 and ICAM1 (with CD11b), chemokines CCL3 and CCL4 (with CCR1), and CSF3 (with CSF3R). Taken together, our results show that mammary infection with E. coli, M. bovis, and Strep uberis culminated in the activation of a conserved core of immune genes and pathways including NET formation.
Reward, motivation and brain imaging in human healthy participants – A narrative review
Authors: Weinstein Aviv M.
Published: March 2023

Over the past 20 years there has been an increasing number of brain imaging studies on the mechanisms underlying reward motivation in humans. This narrative review describes studies on the neural mechanisms associated with reward motivation and their relationships with cognitive function in healthy human participants. The brain’s meso-limbic dopamine reward circuitry in humans is known to control reward-motivated behavior in humans. The medial and lateral Pre-Frontal Cortex (PFC) integrate motivation and cognitive control during decision-making and the dorsolateral PFC (dlPFC) integrates and transmits signals of reward to the mesolimbic and meso-cortical dopamine circuits and initiates motivated behavior. The thalamus and insula influence incentive processing in humans and the motor system plays a role in response to action control. There are reciprocal relationships between reward motivation, learning, memory, imagery, working memory, and attention. The most common method of assessing reward motivation is the monetary incentive delay task (DMRT) and there are several meta-analyses of this paradigm. Genetics modulates motivation reward, and dopamine provides the basis for the interaction between motivational and cognitive control. There is some evidence that male adolescents take more risky decisions than female adolescents and that the lateralization of reward-related DA release in the ventral striatum is confined to men. These studies have implications for our understanding of natural reward and psychiatric conditions like addiction, depression and ADHD. Furthermore, the association between reward and memory can help develop treatment techniques for drug addiction that interfere with consolidation of memory. Finally, there is a lack of research on reward motivation, genetics and sex differences and this can improve our understanding of the relationships between reward, motivation and the brain.
Therapeutic potential of a novel peripherally restricted CB1R inverse agonist on the progression of diabetic nephropathy
Authors: Jacquot Laetitia , Pointeau Océane , Roger-Villeboeuf Célia , Passilly-Degrace Patricia , Belkaid Rim , Regazzoni Isaline , Leemput Julia , Buch Chloé , Demizieux Laurent , Vergès Bruno , Degrace Pascal , Crater Glenn , Jourdan Tony
Published: March 2023

Objective

This study assessed the efficacy of INV-202, a novel peripherally restricted cannabinoid type-1 receptor (CB1R) inverse agonist, in a streptozotocin-induced type-1 diabetes nephropathy mouse model.

Methods

Diabetes was induced in 8-week-old C57BL6/J male mice via intraperitoneal injection of streptozotocin (45 mg/kg/day for 5 days); nondiabetic controls received citrate buffer. Diabetic mice were randomized to 3 groups based on blood glucose, polyuria, and albuminuria, and administered daily oral doses for 28-days of INV-202 at 0.3 or 3 mg/kg or vehicle.

Results

INV-202 did not affect body weight but decreased kidney weight compared with the vehicle group. While polyuria was unaffected by INV-202 treatment, urinary urea (control 30.77 ± 14.93; vehicle 189.81 ± 31.49; INV-202 (0.3 mg/kg) 127.76 ± 20; INV-202 (3 mg/kg) 93.70 ± 24.97 mg/24h) and albumin (control 3.06 ± 0.38; vehicle 850.08 ± 170.50; INV-202 (0.3 mg/kg) 290.65 ± 88.70; INV-202 (3 mg/kg) 111.29 ± 33.47 µg/24h) excretion both decreased compared with vehicle-treated diabetic mice. Compared with the vehicle group, there was a significant improvement in the urinary albumin to creatinine ratio across INV-202 groups. Regardless of the dose, INV-202 significantly reduced angiotensin II excretion in diabetic mice. The treatment also decreased Agtr1a renal expression in a dose-dependent manner. Compared with nondiabetic controls, the glomerular filtration rate was increased in the vehicle group and significantly decreased by INV-202 at 3 mg/kg. While the vehicle group showed a significant loss in the mean number of podocytes per glomerulus, INV-202 treatment limited podocyte loss in a dose-dependent manner. Moreover, in both INV-202 groups, expression of genes coding for podocyte structural proteins nephrin (Nphs1), podocin (Nphs2), and podocalyxin (Pdxl) were restored to levels similar to nondiabetic controls. INV-202 partially limited the proximal tubular epithelial cell (PTEC) hyperplasia and normalized genetic markers for PTEC lesions. INV-202 also reduced expression of genes contributing to oxidative stress (Nox2, Nox4, and P47phox) and inflammation (Tnf). In addition, diabetes-induced renal fibrosis was significantly reduced by INV-202.

Conclusions

INV-202 reduced glomerular injury, preserved podocyte structure and function, reduced injury to PTECs, and ultimately reduced renal fibrosis in a streptozotocin-induced diabetic nephropathy mouse model. These results suggest that INV-202 may represent a new therapeutic option in the treatment of diabetic kidney disease.
Improving the models for prognosis of aneurysmal subarachnoid hemorrhage with the neutrophil-to-albumin ratio
Authors: Zhang Renjie , Liu Zheran , Zhang Yu , Pei Yiyan , He Yan , Yu Jiayi , You Chao , Ma Lu , Fang Fang
Published: March 2023

Objective

Many peripheral inflammatory markers were reported to be associated with the prognosis of aneurysmal subarachnoid hemorrhage (aSAH). We aimed to identify the most promising inflammatory factor that can improve existing predictive models.

Methods

The study was based on data from a 10 year retrospective cohort study at Sichuan University West China Hospital. We selected the well-known SAFIRE and Subarachnoid Hemorrhage International Trialists’ (SAHIT) models as the basic models. We compared the performance of the models after including the inflammatory markers and that of the original models. The developed models were internally and temporally validated.

Results

A total of 3,173 patients were included in this study, divided into the derivation cohort (n = 2,525) and the validation cohort (n = 648). Most inflammatory markers could improve the SAH model for mortality prediction in patients with aSAH, and the neutrophil-to-albumin ratio (NAR) performed best among all the included inflammatory markers. By incorporating NAR, the modified SAFIRE and SAHIT models improved the area under the receiver operator characteristics curve (SAFIRE+NAR vs. SAFIRE: 0.794 vs. 0.778, p = 0.012; SAHIT+NAR vs. SAHIT: 0.831 vs. 0.819, p = 0.016) and categorical net reclassification improvement (SAFIRE+NAR: 0.0727, p = 0.002; SAHIT+NAR: 0.0810, p < 0.001).

Conclusion

This study illustrated that among the inflammatory markers associated with aSAH prognosis, NAR could improve the SAFIRE and SAHIT models for 3 month mortality of aSAH.
Differentiating neurosarcoidosis from multiple sclerosis using combined analysis of basic CSF parameters and MRZ reaction
Authors: Vlad Benjamin , Neidhart Stephan , Hilty Marc , Ziegler Mario , Jelcic Ilijas
Published: March 2023

Background

Neurosarcodosis is one of the most frequent differential diagnoses of multiple sclerosis (MS) and requires central nervous system (CNS) biopsy to establish definite diagnosis according to the latest consensus diagnostic criteria. We here analyzed diagnostic values of basic cerebrospinal fluid (CSF) parameters to distinguish neurosarcoidosis from MS without CNS biopsy.

Methods

We retrospectively assessed clinical, radiological and laboratory data of 27 patients with neurosarcoidosis treated at our center and compared following CSF parameters with those of 138 patients with relapsing-remitting MS: CSF white cell count (WCC), CSF/serum albumin quotient (Qalb), intrathecal production of immunoglobulins including oligoclonal bands (OCB), MRZ reaction, defined as a polyspecific intrathecal production of IgG reactive against ≥2 of 3 the viruses measles (M), rubella (R), and zoster (Z) virus, and CSF lactate levels. Additional inflammatory biomarkers in serum and/or CSF such as neopterin, soluble interleukin-2 receptor (sIL-2R) and C-reactive protein (CRP) were assessed.

Results

There was no significant difference in the frequency of CSF pleocytosis, but a CSF WCC > 30/μl was more frequent in patients with neurosarcoidosis. Compared to MS, patients with neurosarcoidosis showed more frequently an increased Qalb and CSF lactate levels as well as increased serum and CSF levels of sIL-2R, but a lower frequency of intrathecal IgG synthesis and positive MRZ reaction. Positive likelihood ratio (PLR) of single CSF parameters indicating neurosarcoidosis was highest, if (a) CSF WCC was >30/μl (PLR 7.2), (b) Qalb was >10 × 10−3 (PLR 66.4), (c) CSF-specific OCB were absent (PLR 11.5), (d) CSF lactate was elevated (PLR 23.0) or (e) sIL-2R was elevated (PLR>8.0). The combination of (a) one of three following basic CSF parameters, i.e., (a.1.) CSF WCC >30/ul, or (a.2.) QAlb >10 × 10−3, or (a.3.) absence of CSF-specific OCB, and (b) absence of positive MRZ reaction showed the best diagnostic accuracy (sensitivity and specificity each >92%; PLR 12.8 and NLR 0.08).

Conclusion

Combined evaluation of basic CSF parameters and MRZ reaction is powerful in differentiating neurosarcoidosis from MS, with moderate to severe pleocytosis and QAlb elevation and absence of intrathecal IgG synthesis as useful rule-in parameters and positive MRZ reaction as a rule-out parameter for neurosarcoidosis.
CNS efficacy parameters of combination antiretroviral therapy in chronic HIV infection: A multi-voxel magnetic resonance spectroscopy study
Authors: Brkic Snezana , Veres Benjamin , Thurnher Majda M. , Boban Jasmina , Radovanovic Bojan , Tomic Slavica , Kozic Dusko
Published: March 2023

This study aimed to determine the correlations of combination antiretroviral therapy (cART) efficacy parameters in the central nervous system (CNS) with a neurometabolic profile on magnetic resonance spectroscopy (MRS) in virally suppressed, neurologically asymptomatic HIV+ individuals. In total, 32 HIV+ individuals on stable cART with an average age of 41.97 ± 10.12 years and with available clinical data, CNS penetration effectiveness (CPE), and monocyte efficacy (ME) scores underwent multi-voxel MRS. The parameters of neuronal number/function (NAA/Cr), membrane turnover (Cho/Cr), and glial proliferation (mI/Cr) were analyzed in supratentorial white and gray matter. Correlations of CPE and ME with neurometabolic ratios were performed using Pearson's correlation test. Statistical significance was set at p < 0.05. A strong positive correlation was observed between Cho/Cr and CPE in the left parietal subcortical white matter (r = 0.577, p = 0.001). A strong positive correlation between NAA/Cr and ME was obtained in the left (r = 0.521, p = 0.003) and the right (r = 0.494, p = 0.005) posterior cingulate. A strong negative correlation between ME and Cho/Cr ratios was observed in the right frontal deep white matter (r = −0.569, p = 0.001). Indices designed to assess cART efficacy in CNS failed to present significant correlations with the neurometabolic profile obtained using MRS. There is a need to define more potent non-invasive tools for neuroinflammation assessment given the prolonged life expectancy in the HIV+ population.
Non-stationary neural signal to image conversion framework for image-based deep learning algorithms
Authors: Patel Sahaj Anilbhai , Yildirim Abidin
Published: March 2023

This paper presents a time-efficient preprocessing framework that converts any given 1D physiological signal recordings into a 2D image representation for training image-based deep learning models. The non-stationary signal is rasterized into the 2D image using Bresenham’s line algorithm with time complexity O(n). The robustness of the proposed approach is evaluated based on two publicly available datasets. This study classified three different neural spikes (multi-class) and EEG epileptic seizure and non-seizure (binary class) based on shapes using a modified 2D Convolution Neural Network (2D CNN). The multi-class dataset consists of artificially simulated neural recordings with different Signal-to-Noise Ratios (SNR). The 2D CNN architecture showed significant performance for all individual SNRs scores with (SNR/ACC): 0.5/99.69, 0.75/99.69, 1.0/99.49, 1.25/98.85, 1.5/97.43, 1.75/95.20 and 2.0/91.98. Additionally, the binary class dataset also achieved 97.52% accuracy by outperforming several others proposed algorithms. Likewise, this approach could be employed on other biomedical signals such as Electrocardiograph (EKG) and Electromyography (EMG).
Prolonged maternal exposure to glucocorticoids alters selenoprotein expression in the developing brain
Authors: Toh Pamela , Seale Lucia A. , Berry Marla J. , Torres Daniel J.
Published: March 2023

Aberrant activation of the stress-response system in early life can alter neurodevelopment and cause long-term neurological changes. Activation of the hypothalamic–pituitary–adrenal axis releases glucocorticoids into the bloodstream, to help the organism adapt to the stressful stimulus. Elevated glucocorticoid levels can promote the accumulation of reactive oxygen species, and the brain is highly susceptible to oxidative stress. The essential trace element selenium is obtained through diet, is used to synthesize antioxidant selenoproteins, and can mitigate glucocorticoid-mediated oxidative damage. Glucocorticoids can impair antioxidant enzymes in the brain, and could potentially influence selenoprotein expression. We hypothesized that exposure to high levels of glucocorticoids would disrupt selenoprotein expression in the developing brain. C57 wild-type dams of recently birthed litters were fed either a moderate (0.25 ppm) or high (1 ppm) selenium diet and administered corticosterone (75 μg/ml) via drinking water during postnatal days 1 to 15, after which the brains of the offspring were collected for western blot analysis. Glutathione peroxidase 1 and 4 levels were increased by maternal corticosterone exposure within the prefrontal cortex, hippocampus, and hypothalamus of offspring. Additionally, levels of the glucocorticoid receptor were decreased in the hippocampus and selenoprotein W was elevated in the hypothalamus by corticosterone. Maternal consumption of a high selenium diet independently decreased glucocorticoid receptor levels in the hippocampus of offspring of both sexes, as well as in the prefrontal cortex of female offspring. This study demonstrates that early life exposure to excess glucocorticoid levels can alter selenoprotein levels in the developing brain.
Opening a window to the acutely injured brain: Simultaneous retinal and cerebral vascular monitoring in rats
Authors: Warner Laura , Bach-Hagemann Annika , Schmidt Tobias P. , Pinkernell Sarah , Schubert Gerrit A. , Clusmann Hans , Albanna Walid , Lindauer Ute , Conzen-Dilger Catharina
Published: March 2023

Many recent research projects have described typical chronic changes in the retinal vasculature for diverse neurovascular and neurodegenerative disorders such as stroke or Alzheimer's disease. Unlike cerebral vasculature, retinal blood vessels can be assessed non-invasively by retinal vessel analysis. To date, there is only a little information about potential simultaneous reactions of retinal and cerebral vessels in acute neurovascular diseases. The field of applications of retinal assessment could significantly be widened if more information about potential correlations between those two vascular beds and the feasibility of non-invasive retinal vessel analysis in acute neurovascular disease were available. Here, we present our protocol for the simultaneous assessment of retinal and cerebral vessels in an acute setting in anesthetized rats using a non-invasive retinal vessel analyzer and a superficial tissue imaging system for laser speckle contrast analysis via a closed bone window. We describe the experimental set-up in detail, outline the pitfalls of repeated retinal vessel analyses in an experimental set-up of several hours, and address issues that arise from the simultaneous use of two different assessment tools. Finally, we demonstrate the robustness and variability of the reactivity of retinal vessels to hypercapnia at baseline as well as their reproducibility over time using two anesthetic protocols common for neurovascular research. In summary, the procedures described in this protocol allow us to directly compare retinal and cerebral vascular beds and help to substantiate the role of the retina as a “window to the brain.”