bluuebunny/crossref_metadata_2025 huggingface dataset:
Development and performance analysis of a ZigBee and LoRa-based smart building sensor network
Authors: Ali Ayanle Ibrahim , Zorlu Partal Sibel
Published: August 2022

A wireless sensor network employing ZigBee and LoRa (Long Range) communication protocols for integration into smart building energy management systems is presented in this article. The design and implementation details are provided, and the performance parameters of the communication network are defined and analyzed based on the test results obtained from different configurations. The developed embedded system can be used in smart environments so that the room temperature, humidity, lighting systems, and so on can be automatically monitored and controlled. By customizing the embedded code, a variety of Internet of Things (IoT) applications can be introduced owing to their scalability. Taking advantage of the complementing low-power and long-range features of ZigBee and LoRa communication technologies, a system comprising an end device, a multi-protocol gateway, and a central data collector (CDC) unit is developed. The end device collects temperature and humidity as well as light intensity data using low-power sensors and sends the data to the gateway via the LoRa wireless transceiver module. The gateway was designed as an intermediate device that allows data exchange between the LoRa and ZigBee transceiver modules. It receives sensor data from the end device via LoRa and sends them to the CDC unit via a ZigBee-based XBee S2 commercial wireless transceiver module. Sensor data are monitored in the CDC unit by using an open-source IoT software platform. A commercial STM32 integrated circuit (IC) was used as a microcontroller unit for the end device and gateway. Performance parameters such as communication range and throughput data were studied for both the ZigBee and LoRa wireless transceiver modules.
Effect of Dual-Flow Channel Structures on Electrochemical CO2 Reduction in Proton Exchange Membrane Electrolyzers
Authors: Na Youngseung , Ha Min Gwan , Park Hyun S. , Park Hee Young , Kim Hyoung-Juhn , Henkensmeier Dirk , Yoo Sung Jong , Kim Jin Young , Lee So Young , Jang Jong Hyun
Published: August 2022

Greenhouse gases such as carbon dioxide and methane are responsible for intensifying global warming. Consequently, a reduction in power plant outputs and an increase in capture and storage on-site are required to reduce greenhouse gas emissions. Recently, research has focused on an electrochemical CO2 reduction method because the amount of CO2 reduction can be controlled by adjusting the operating voltage. However, to scale up the electrochemical system while maintaining a high conversion rate in a large cell, a suitable flow field of the cell must be optimized. The transparent cell structure presented in this study allows visualization of the distribution of the two-phase flow. Accordingly, dual-flow channels consisting of main and sub-channels have been designed. Furthermore, multiple configurations of the dual-flow channels and locations of the catalyst layer have been compared. The interdigitated sub-channels and inverted layered cell structures can supply gas and liquid to the catalyst layer via distinct pathways, allowing for uniform flow distribution to each channel.
Ecological–Health Risk of Antimony and Arsenic in Centella asiatica, Topsoils, and Mangrove Sediments: A Case Study of Peninsular Malaysia
Authors: Yap Chee Kong , Tan Wen Siang , Cheng Wan Hee , Syazwan Wan Mohd , Azrizal-Wahid Noor , Krishnan Kumar , Go Rusea , Nulit Rosimah , Ibrahim Mohd. Hafiz , Mustafa Muskhazli , Omar Hishamuddin , Chew Weiyun , Edward Franklin Berandah , Okamura Hideo , Al-Mutairi Khalid Awadh , Al-Shami Salman Abdo , Sharifinia Moslem , Keshavarzifard Mehrzad , You Chen Feng , Bakhtiari Alireza Riyahi , Bintal Amin , Zakaly Hesham M. H. , Arai Takaomi , Naji Abolfazl , Saleem Muhammad , Abd Rahman Mohd Amiruddin , Ong Ghim Hock , Subramaniam Geetha , Wong Ling Shing
Published: August 2022

The current study assessed the ecological–health risks of potentially toxic arsenic (As) and antimony (Sb) in the vegetable Centella asiatica, topsoils, and mangrove sediments sampled from Peninsular Malaysia. The As concentrations ranged from 0.21 to 4.33, 0.18 to 1.83, and 1.32 to 20.8 mg/kg dry weight, for the leaves, stems, and roots of the vegetable, respectively. The ranges of Sb concentrations were 0.31–0.62, 0.12–0.35, and 0.64–1.61 mg/kg dry weight, for leaves, stems, and roots of the vegetable, respectively. The children’s target hazard quotient (THQ) values indicated no non-carcinogenic risks of As and Sb in both leaves and stems, although children’s THQ values were higher than those in adults. The calculated values of estimated weekly intake were lower than established provisional tolerable weekly intake of As and Sb for both children and adult consumers. The carcinogenic risk (CR) values of As for children’s intake of leaves and stems of vegetables showed more public concern than those of adults. The levels of Sb and As in the topsoils were generally higher (although not significantly) than those in the mangrove sediments, resulting in a higher geoaccumulation index, contamination factor and ecological risk, hazard index, THQ, and CR values. This indicated that the anthropogenic sources of Sb and As originated from the land-based activities before reaching the mangrove near the coast. The CR of As signifies a dire need for comprehensive ecological–health risks exposure studies, as dietary intake involves more than just vegetable consumption. Therefore, risk management for As and Sb in Malaysia is highly recommended. The present findings of the ecological–health risks of As and Sb based on 2010–2012 samples can be used as an important baseline for future reference and comparison.
Contrasting corn yield responses to nitrogen fertilization in southeast coastal plain soils
Authors: Raza Sajjad , Farmaha Bhupinder Singh
Published: August 2022

Nitrogen (N) fertilizer recommendations based on yield goals are common and do not consider variability in soil N supply between fields. Nitrogen transformations in soils are dynamic and may vary widely among different soils, climates, cropping systems, and management practices, making it difficult to provide general N recommendation rates for a region, state, or even a county. To optimize N fertilization rates for corn (Zea mays L.), eight field trials were conducted on different fields at the Edisto Research and Education Center of Clemson University in 2018–2021. The soils varied in pH, inorganic N, management practices (irrigate, dryland, cover, and no-cover crop), and climatic conditions (temperature and precipitation). Six nitrogen fertilizer rate treatments were used during 2018 and 2019, eight in 2020, and five in 2021 trials. Test sites were different in each year except in 2021 in which trials were conducted on the same site but were differentiated by multi-species and no-cover crop treatments. Nitrogen fertilizer was applied as a complete one-time dose in all trials in 2018 and 2021 and in several splits in 2019 and 2020. The corn yields generally increased with N fertilization rates; however, a strong variation of up to 4–11 Mg ha−1 existed at each N level among different fields. Based on yield responses, the eight field trials were divided into two groups: low-yielding and high-yielding sites. The low-yielding sites on average produced 3.8 Mg ha−1 of corn, which was 137% less than the corn harvested from high-yielding sites (9.0 Mg ha−1). The agronomic N use efficiency (AgNUE) in high-yielding sites was almost double (32.68 kg kg−1 N) compared to low-yielding sites (17.16 kg kg−1 N), and the differences were even wider for partial factor productivity of N (PFPN). The economic optimum N rate (EONR) remained below 300 kg N ha−1 for all sites with no marked difference among sites. However, relative yield (ratio of corn yield in unfertilized control to the N level giving the highest yield) was comparatively lower in low-yielding sites (29.45%). Baseline inorganic N, pH, and precipitation were identified as the major factors controlling corn yield responses to N fertilization. The yield variations driven by several factors make N management challenging, indicating the need for site-specific N management for corn in the southeast United States.
Evolution of the odorant-binding protein gene family in Drosophila
Authors: Rondón Johnma J. , Moreyra Nicolás Nahuel , Pisarenco Vadim A. , Rozas Julio , Hurtado Juan , Hasson Esteban
Published: August 2022

Odorant-binding proteins (OBPs) are encoded by a gene family involved in the perception of olfactory signals in insects. This chemosensory gene family has been advocated as a candidate to mediate host preference and host shifts in insects, although it also participates in other physiological processes. Remarkable differences in the OBP gene repertoire have been described across insect groups, suggesting an accelerated gene turnover rate. The genus Drosophila, is a valuable resource for ecological genomics studies since it comprises groups of ecologically diverse species and there are genome data for many of them. Here, we investigate the molecular evolution of this chemosensory gene family across 19 Drosophila genomes, including the melanogaster and repleta species groups, which are mostly associated with rotting fruit and cacti, respectively. We also compared the OBP repertoire among the closely related species of the repleta group, associated with different subfamilies of Cactaceae that represent disparate chemical challenges for the flies. We found that the gene family size varies widely between species, ranging from 39 to 54 candidate OBPs. Indeed, more than 54% of these genes are organized in clusters and located on chromosomes X, 2, and 5, with a distribution conserved throughout the genus. The family sizes in the repleta group and D. virilis (virilis-repleta radiation) were smaller than in the melanogaster group. We tested alternative evolutionary models for OBP family size and turnover rates based on different ecological scenarios. We found heterogeneous gene turnover rates (GR) in comparisons involving columnar cactus specialists, prickly pear specialists, and fruit dwellers lineages, and signals of rapid molecular evolution compatible with positive selection in specific OBP genes. Taking ours and previous results together, we propose that this chemosensory gene family is involved in host adaptation and hypothesize that the adoption of the cactophilic lifestyle in the repleta group accelerated the evolution of members of the family.
Food web functional responses
Authors: Abrams Peter A.
Published: August 2022

This article reviews the nature of functional responses that have commonly been used to represent feeding relationships in the ecological literature. It compares these with the range of functional response forms that are likely to characterize species in natural communities. The latter set of responses involves many more variables. The article reviews the history of functional response models, and examines previous work that has allowed the functional response of a predator to a single type of prey to depend on additional variables beyond the abundance of that prey type. While a number of more complex functional responses have been discussed over the years, many variables affecting feeding rates are still typically omitted from models of food webs. The influences on functional responses from trophic levels above that of the predator or below that of the prey are particularly likely to be ignored, although models and data have suggested that they can have large effects on the functional response. The influences of adaptive behavior and of the time-scale of response measurement are also too often ignored. Some of the known and unknown consequences of these omissions are discussed.
Conservation status and historical relatedness of South African communal indigenous goat populations using a genome-wide single-nucleotide polymorphism marker
Authors: Chokoe T. C. , Hadebe K. , Muchadeyi F. C. , Nephawe K. A. , Dzomba E. F. , Mphahlele T. D. , Matelele T. C. , Mtileni B. J.
Published: August 2022

Indigenous goats form the majority of populations in smallholder, low input, low output production systems and are considered an important genetic resource due to their adaptability to different production environments and support of communal farming. Effective population size (Ne), inbreeding levels, and the runs of homozygosity (ROHs) are effective tools for exploring the genetic diversity and understanding the demographic history in efforts to support breeding strategies to use and conserve genetic resources. Across populations, the current Ne of Gauteng was the lowest at 371 animals, while the historical Ne across populations suggests that the ancestor Ne has decreased by 53.86%, 44.58%, 42.16%, and 41.16% in Free State (FS), North West (NW), Limpopo (LP), and Gauteng (GP), respectively, over the last 971 generations. Genomic inbreeding levels related to ancient kinship (FROH > 5 Mb) were highest in FS (0.08 ± 0.09) and lowest in the Eastern Cape (EC) (0.02 ± 0.02). A total of 871 ROH island regions which include important environmental adaptation and hermo-tolerance genes such as IL10RB, IL23A, FGF9, IGF1, EGR1, MTOR, and MAPK3 were identified (occurring in over 20% of the samples) in FS (n = 37), GP (n = 42), and NW (n = 2) populations only. The mean length of ROH across populations was 7.76 Mb and ranged from 1.61 Mb in KwaZulu-Natal (KZN) to 98.05 Mb (GP and NW). The distribution of ROH according to their size showed that the majority (n = 1949) of the detected ROH were > 5 Mb in length compared to the other categories. Assuming two hypothetical ancestral populations, the populations from KZN and LP are revealed, supporting PC 1. The genomes of KZN and LP share a common origin but have substantial admixture from the EC and NW populations. The findings revealed that the occurrence of high Ne and autozygosity varied largely across breeds in communal indigenous goat populations at recent and ancient events when a genome-wide single-nucleotide polymorphism (SNP) marker was used. The use of Illumina goat SNP50K BeadChip shows that there was a migration route of communal indigenous goat populations from the northern part (LP) of South Africa to the eastern areas of the KZN that confirmed their historical relatedness and coincides with the migration periods of the Bantu nation.
Identification and Analysis of Long Non-Coding RNAs Related to UV-B-Induced Anthocyanin Biosynthesis During Blood-Fleshed Peach (Prunus persica) Ripening
Authors: Zhang Man , Zhang Xiuqi , Wang Haijing , Ye Mao , Liu Yating , Song Zhihua , Du Tingting , Cao Hongyan , Song Liqin , Xiao Xiao , Liu Jianzhen , Zhang Libin , Song Yangbo , Yang Qing , Meng Dong , Wu Junkai
Published: August 2022

Blood flesh is a key fruit trait in peaches (Prunus persica) and can be attributed to the accumulation of anthocyanins. The roles of long non-coding RNAs (lncRNAs) have been highlighted by multiple studies in regulating fruit ripening, anthocyanin accumulation, and abiotic stress responses in many flowering plants. Such regulatory functions of lncRNAs in Prunus persica, nonetheless, have not been reported. In this research, we sequenced and analyzed the complete transcriptome of C3-20 (a blood-fleshed peach) fruit at four developmental stages. Analyses of the correlated genes and differentially expressed lncRNA target genes helped to forecast lncRNAs’ possible functions. The RNA-seq data were generated using high-throughput sequencing. In total, 17,456 putative lncRNAs, including 4,800 intergenic lncRNAs, 2,199 antisense lncRNAs, and 10,439 intronic lncRNAs were discovered, of which 4,871 differentially expressed lncRNAs (DE-lncRNAs) were annotated in the fruit developmental processes. The target genes of these DE-lncRNAs and their regulatory relationship identifying 21,795 cis-regulated and 18,271 trans-regulated targets of the DE-lncRNAs were in a similar way predicted by us. The enriched GO terms for the target genes included anthocyanin biosynthesis. Flavonoid biosynthesis and plant hormone signal transduction were also included in the enriched KEGG pathways. Co-expression network construction demonstrated that the highly expressed genes might co-regulate multiple other genes associated with auxin signal transduction and take effect in equal pathways. We discovered that lncRNAs, including LNC_000987, LNC_000693, LNC_001323, LNC_003610, LNC_001263, and LNC_003380, correlated with fruit that ripened and could take part in ethylene biosynthesis and metabolism and the ABA signaling pathway. Several essential transcription factors, such as ERFs, WRKY70, NAC56, and NAC72, may in a similar way regulate fruit ripening. Three DE-lncRNAs, XLOC_011933, XLOC_001865, and XLOC_042291, are involved in UV-B-induced anthocyanin biosynthesis and positively regulating UVR8 and COP10, were identified and characterized. Our discovery and characterization of XLOC_011933, XLOC_001865, and XLOC_042291 provide a more precise understanding and preliminarily establishes a theoretical framework for UV-B-induced flesh anthocyanin biosynthesis. This phenomenon might encourage more in-depth investigations to study the molecular mechanisms underlying peach flesh coloring.
B-cell response in solid organ transplantation
Authors: Yi Stephanie G. , Gaber Ahmed Osama , Chen Wenhao
Published: August 2022

The transcriptional regulation of B-cell response to antigen stimulation is complex and involves an intricate network of dynamic signals from cytokines and transcription factors propagated from T-cell interaction. Long-term alloimmunity, in the setting of organ transplantation, is dependent on this B-cell response, which does not appear to be halted by current immunosuppressive regimens which are targeted at T cells. There is emerging evidence that shows that B cells have a diverse response to solid organ transplantation that extends beyond plasma cell antibody production. In this review, we discuss the mechanistic pathways of B-cell activation and differentiation as they relate to the transcriptional regulation of germinal center B cells, plasma cells, and memory B cells in the setting of solid organ transplantation.
FYN regulates cell adhesion at the blood-testis barrier and the apical ectoplasmic specialization via its effect on Arp3 in the mouse testis
Authors: Yang Yue , Yao Mingxia , Zeng Jie , Zheng Dongwang , Li Qin , Ni Ya , Xiao Xiang
Published: August 2022

FYN is a non-receptor tyrosine kinase of the SRC family that facilitates virus entry across epithelial tight junctions. However, the role of FYN in mammalian testes in maintaining the blood-testis barrier (BTB) integrity and the adhesion of germ cells to Sertoli cells are not well defined. Here, we show that FYN is a component of the BTB and the apical ectoplasmic specialization (ES) at Sertoli-Sertoli and Sertoli-spermatid interfaces, respectively, and is expressed extensively in mouse testes during postnatal development. FYN was shown to be structurally linked to the actin and microtubule-based cytoskeletons. An in vivo model was used to explore the modulatory effect of FYN on BTB and apical ES dynamics within the testes when adult mice were treated intraperitoneally with CdCl2 (3 mg/kg body weight). The CdCl2-induced epithelial restructuring was associated with a transient increase in the interaction between FYN and the actin branching/nucleation protein Arp3, as well as an induction of Arp3 phosphorylation, which possibly lead to actin cytoskeleton remodeling, resulting in BTB damage and germ cell loss in the seminiferous epithelium. Based on the results, we propose a model in which FYN and Arp3 form a protein complex that is responsible for junction reorganization events at the apical ES and the BTB. It is also possible for viruses to break through the BTB and enter the immunoprivileged testicular microenvironment via this mechanism.
“Iron triangle” of regulating the uterine microecology: Endometrial microbiota, immunity and endometrium
Authors: Zhu Na , Yang Xuyan , Liu Qiao , Chen Yahui , Wang Xiaolan , Li Huanhuan , Gao Hong
Published: August 2022

The uterus is the core place for breeding new life. The balance and imbalance of uterine microecology can directly affect or even dominate the female reproductive health. Emerging data demonstrate that endometrial microbiota, endometrium and immunity play an irreplaceable role in regulating uterine microecology, forming a dynamic iron triangle relationship. Up to nowadays, it remains unclear how the three factors affect and interact with each other, which is also a frontier topic in the emerging field of reproductive tract microecology. From this new perspective, we aim to clarify the relationship and mechanism of the interaction of these three factors, especially their pairwise interactions. Finally, the limitations and future perspectives of the current studies are summarized. In general, these three factors have a dynamic relationship of mutual dependence, promotion and restriction under the physiological or pathological conditions of uterus, among which the regulatory mechanism of microbiota and immunity plays a role of bridge. These findings can provide new insights and measures for the regulation of uterine microecology, the prevention and treatment of endometrial diseases, and the further multi-disciplinary integration between microbiology, immunology and reproductive medicine.
The dark side of Tregs during aging
Authors: Palatella Martina , Guillaume Stephane M. , Linterman Michelle A. , Huehn Jochen
Published: August 2022

In the last century, we have seen a dramatic rise in the number of older persons globally, a trend known as the grey (or silver) tsunami. People live markedly longer than their predecessors worldwide, due to remarkable changes in their lifestyle and in progresses made by modern medicine. However, the older we become, the more susceptible we are to a series of age-related pathologies, including infections, cancers, autoimmune diseases, and multi-morbidities. Therefore, a key challenge for our modern societies is how to cope with this fragile portion of the population, so that everybody could have the opportunity to live a long and healthy life. From a holistic point of view, aging results from the progressive decline of various systems. Among them, the distinctive age-dependent changes in the immune system contribute to the enhanced frailty of the elderly. One of these affects a population of lymphocytes, known as regulatory T cells (Tregs), as accumulating evidence suggest that there is a significant increase in the frequency of these cells in secondary lymphoid organs (SLOs) of aged animals. Although there are still discrepancies in the literature about modifications to their functional properties during aging, mounting evidence suggests a detrimental role for Tregs in the elderly in the context of bacterial and viral infections by suppressing immune responses against non-self-antigens. Interestingly, Tregs seem to also contribute to the reduced effectiveness of immunizations against many pathogens by limiting the production of vaccine-induced protective antibodies. In this review, we will analyze the current state of understandings about the role of Tregs in acute and chronic infections as well as in vaccination response in both humans and mice. Lastly, we provide an overview of current strategies for Treg modulation with potential future applications to improve the effectiveness of vaccines in older individuals.
Substratum interactions determine immune response to allogeneic transplants of endothelial cells
Authors: Wilcox Elise C. , Edelman Elazer R.
Published: August 2022

Endothelial cells (ECs) are central to vascular health but also interact with and regulate the immune system. Changes in endothelial state enable immune cells to migrate into the tissue to facilitate repair and fight infection. ECs modulate the function of immune cells through the expression of adhesion molecules, chemokines, major histocompatibility complex (MHC), and an array of co-stimulatory and inhibitor molecules. These interactions allow ECs to act as antigen presenting cells (APCs) and influence the outcome of immune recognition. This study elucidates how EC microenvironment, vascular cell biology, and immune response are not only connected but interdependent. More specifically, we explored how cell-substratum interactions influence EC antigen presentation and co-stimulation, and how these differences affect allorecognition in animal models of cell transplantation. Investigation of EC state was carried out using RNA sequencing while assessment of the allogeneic response includes measurements of immune cell cytotoxic ability, T cell proliferation, cytokine release, serum antibodies, and histological staining. Differences in substratum led to divergent EC phenotypes which in turn influenced immune response to transplanted cells, both due to the physical barrier of matrix-adhesion and differences in expression of surface markers. ECs grown in 2D on tissue culture plastic or in 3D on collagen scaffolds had significantly different basal levels of MHC expression, co-stimulatory and adhesion molecules. When treated with cytokines to mimic an inflammatory state, ECs did not converge to a single phenotype but rather responded differently based on their substratum. Generally, 3D ECs were more responsive to inflammatory stimuli than 2D ECs. These unique expression patterns measuredin vitroalso influence immune recognitionin vivo. ECs grown in 2D were more likely to provoke a cytotoxic response while 3D ECs induced T cell proliferation. ECs are uniquely configured to sense not only local flow and mechanical forces but a range of markers related to systemic state, including immune function. ECs interact with immune cells with differing results depending on the environment in which the EC-lymphocyte interaction occurs. Therefore, understanding this relationship is essential to predicting and modifying the outcome of EC-immune interacts. We specifically examined the relationship between EC substratum and allorecognition.
IL-10 family cytokines in chronic rhinosinusitis with nasal polyps: From experiments to the clinic
Authors: Xuan Lijia , Zhang Nan , Wang Xiangdong , Zhang Luo , Bachert Claus
Published: August 2022

Chronic rhinosinusitis with nasal polyps (CRSwNP) is considered a nasal sinus inflammatory disease that can be dominated by immune cells and cytokines. IL-10 family cytokines exert essential functions in immune responses during infection and inflammation. Recently, the understanding of the roles of the IL-10 family in CRSwNP is being reconsidered. IL-10 family members are now considered complex cytokines that are capable of affecting epithelial function and involved in allergies and infections. Furthermore, the IL-10 family responds to glucocorticoid treatment, and there have been clinical trials of therapies manipulating these cytokines to remedy airway inflammatory diseases. Here, we summarize the recent progress in the understanding of IL-10 family cytokines in CRSwNP and suggest more specific strategies to exploit these cytokines for the effective treatment of CRSwNP.
Refining the DC-targeting vaccination for preventing emerging infectious diseases
Authors: Pastor Yadira , Ghazzaui Nour , Hammoudi Adele , Centlivre Mireille , Cardinaud Sylvain , Levy Yves
Published: August 2022

The development of safe, long-term, effective vaccines is still a challenge for many infectious diseases. Thus, the search of new vaccine strategies and production platforms that allow rapidly and effectively responding against emerging or reemerging pathogens has become a priority in the last years. Targeting the antigens directly to dendritic cells (DCs) has emerged as a new approach to enhance the immune response after vaccination. This strategy is based on the fusion of the antigens of choice to monoclonal antibodies directed against specific DC surface receptors such as CD40. Since time is essential,in silicoapproaches are of high interest to select the most immunogenic and conserved epitopes to improve the T- and B-cells responses. The purpose of this review is to present the advances in DC vaccination, with special focus on DC targeting vaccines and epitope mapping strategies and provide a new framework for improving vaccine responses against infectious diseases.
Road testing new CAR design strategies in multiple myeloma
Authors: Rana Priyanka S. , Murphy Elena V. , Kort Jeries , Driscoll James J.
Published: August 2022

A deeper understanding of basic immunology principles and advances in bioengineering have accelerated the mass production of genetically-reprogrammed T-cells as living drugs to treat human diseases. Autologous and allogeneic cytotoxic T-cells have been weaponized to brandish MHC-independent chimeric antigen receptors (CAR) that specifically engage antigenic regions on tumor cells. Two distinct CAR-based therapeutics designed to target BCMA are now FDA-approved based upon robust, sustained responses in heavily-pretreated multiple myeloma (MM) patients enrolled on the KarMMa and CARTITUDE-1 studies. While promising, CAR T-cells present unique challenges such as antigen escape and T-cell exhaustion. Here, we review novel strategies to design CARs that overcome current limitations. Co-stimulatory signaling regions were added to second-generation CARs to promote IL-2 synthesis, activate T-cells and preclude apoptosis. Third-generation CARs are composed of multiple co-stimulatory signaling units, e.g., CD28, OX40, 4-1BB, to reduce exhaustion. Typically, CAR T-cells incorporate a potent constitutive promoter that maximizes long-term CAR expression but extended CAR activation may also promote T-cell exhaustion. Hypoxia-inducible elements can be incorporated to conditionally drive CAR expression and selectively target MM cells within bone marrow. CAR T-cell survival and activity is further realized by blocking intrinsic regulators of T-cell inactivation. T-Cells Redirected for Universal Cytokine Killing (TRUCKs) bind a specific tumor antigen and produce cytokines to recruit endogenous immune cells. Suicide genes have been engineered into CAR T-cells given the potential for long-term on-target, off-tumor effects. Universal allo-CAR T-cells represent an off-the-shelf source, while logic-gated CAR T-cells are designed to recognize tumor-specific features coupled with Boolean-generated binary gates that then dictate cell-fate decisions. Future generations of CARs should further revitalize immune responses, enhance tumor specificity and reimagine strategies to treat myeloma and other cancers.
Insights into the role of neutrophils in neuropsychiatric systemic lupus erythematosus: Current understanding and future directions
Authors: Sim Tao Ming , Mak Anselm , Tay Sen Hee
Published: August 2022

Central nervous system (CNS) involvement of systemic lupus erythematosus (SLE), termed neuropsychiatric SLE (NPSLE), is a major and debilitating manifestation of the disease. While patients with SLE mostly complain of common neuropsychological symptoms such headache and mild mood disorders that may not even be technically attributed to SLE, many SLE patients present with life-threatening NPSLE syndromes such as cerebrovascular disease, seizures and psychosis that are equally challenging in terms of early diagnosis and therapy. While we are just beginning to unravel some mysteries behind the immunologic basis of NPSLE, advancements in the mechanistic understanding of the complex pathogenic processes of NPSLE have been emerging through recent murine and human studies. The pathogenic pathways implicated in NPSLE are multifarious and various immune effectors such as cell-mediated inflammation, autoantibodies and cytokines including type I interferons have been found to act in concert with the disruption of the blood-brain barrier (BBB) and other neurovascular interfaces. Beyond antimicrobial functions, neutrophils are emerging as decision-shapers during innate and adaptive immune responses. Activated neutrophils have been recognized to be involved in ischemic and infective processes in the CNS by releasing neutrophil extracellular traps (NETs), matrix metalloproteinase-9 and proinflammatory cytokines. In the context of NPSLE, these mechanisms contribute to BBB disruption, neuroinflammation and externalization of modified proteins on NETs that serve as autoantigens. Neutrophils that sediment within the peripheral blood mononuclear cell fraction after density centrifugation of blood are generally defined as low-density neutrophils (LDNs) or low-density granulocytes. LDNs are a proinflammatory subset of neutrophils that are increased with SLE disease activity and are primed to undergo NETosis and release cytokines such as interferon-α and tumor necrosis factor. This review discusses the immunopathogenesis of NPSLE with a focus on neutrophils as a core mediator of the disease and potential target for translational research in NPSLE.
Enhanced immunogenicity of a positively supercharged archaeon thioredoxin scaffold as a cell-penetrating antigen carrier for peptide vaccines
Authors: Cavazzini Davide , Spagnoli Gloria , Mariz Filipe Colaco , Reggiani Filippo , Maggi Stefano , Franceschi Valentina , Donofrio Gaetano , Müller Martin , Bolchi Angelo , Ottonello Simone
Published: August 2022

Polycationic resurfaced proteins hold great promise as cell-penetrating bioreagents but their use as carriers for the intracellular delivery of peptide immuno-epitopes has not thus far been explored. Here, we report on the construction and functional characterization of a positively supercharged derivative of Pyrococcus furiosus thioredoxin (PfTrx), a thermally hyperstable protein we have previously validated as a peptide epitope display and immunogenicity enhancing scaffold. Genetic conversion of 13 selected amino acids to lysine residues conferred to PfTrx a net charge of +21 (starting from the -1 charge of the wild-type protein), along with the ability to bind nucleic acids. In its unfused form, +21 PfTrx was readily internalized by HeLa cells and displayed a predominantly cytosolic localization. A different intracellular distribution was observed for a +21 PfTrx-eGFP fusion protein, which although still capable of cell penetration was predominantly localized within endosomes. A mixed cytosolic/endosomal partitioning was observed for a +21 PfTrx derivative harboring three tandemly repeated copies of a previously validated HPV16-L2 (aa 20-38) B-cell epitope grafted to the display site of thioredoxin. Compared to its wild-type counterpart, the positively supercharged antigen induced a faster immune response and displayed an overall superior immunogenicity, including a substantial degree of self-adjuvancy. Altogether, the present data point to +21 PfTrx as a promising novel carrier for intracellular antigen delivery and the construction of potentiated recombinant subunit vaccines.
PD-L1+ neutrophils as novel biomarkers for stage IV melanoma patients treated with nivolumab
Authors: Cristinziano Leonardo , Modestino Luca , Capone Mariaelena , Madonna Gabriele , Mallardo Domenico , Giannarelli Diana , D’Angelo Grazia , Ferrara Anne Lise , Loffredo Stefania , Varricchi Gilda , Vanella Vito , Festino Lucia , Ascierto Paolo Antonio , Galdiero Maria Rosaria
Published: August 2022

Melanoma displays a rising incidence, and the mortality associated with metastatic form remains high. Monoclonal antibodies that block programmed death (PD-1) and PD Ligand 1 (PD-L1) network have revolutionized the history of metastatic disease. PD-L1 is expressed on several immune cells and can be also expressed on human neutrophils (PMNs). The role of peripheral blood PMNs as predictive biomarkers in anti-PD-1 therapy of melanoma is largely unknown. In this study, we aimed to determine activation status and PD-L1 expression on human neutrophils as possible novel biomarkers in stage IV melanoma patients (MPs). We found that PMNs from MPs displayed an activated phenotype and increased PD-L1 levels compared to healthy controls (HCs). Patients with lower PD-L1+ PMN frequencies displayed better progression-free survival (PFS) and overall survival (OS) compared to patients with high PD-L1+ PMN frequencies. Multivariate analysis showed that PD-L1+ PMNs predicted patient outcome in BRAF wild type MP subgroup but not in BRAF mutated MPs. PD-L1+ PMN frequency emerges as a novel biomarker in stage IV BRAF wild type MPs undergoing anti-PD-1 immunotherapy. Our findings suggest further evaluation of the role of neutrophil subsets and their mediators in melanoma patients undergoing immunotherapy.
Time-course transcriptome analyses of spleen in rainbow trout (Oncorhynchus mykiss) post-Flavobacterium psychrophilum infection
Authors: Deng Furong , Wang Di , Loch Thomas P. , Chen Fuguang , Lu Tongyan , Cao Yongsheng , Fan Dan , Li Shaowu
Published: August 2022

Flavobacterium psychrophilum, the etiological agent of bacterial coldwater disease and rainbow trout fry syndrome, causes considerable losses in salmonid aquaculture globally. Systemic F. psychrophilum infections in rainbow trout (Oncorhynchus mykiss) lead to a range of clinical signs, including ulcerative lesions in the skin and muscle and splenitis. Previous studies offered an integrative analysis of the skeletal muscle response to F. psychrophilum infection in rainbow trout. However, little is known about the molecular mechanism of immune response in the spleen, which is an important immune organ of rainbow trout. Here, we investigated the time-course splenic transcriptome profiles in uninfected rainbow trout (CK) and F. psychrophilum–infected rainbow trout at day 3 and day 7 (D3, D7) by RNA-seq analyses. Among the 7,170 differentially expressed genes (DEGs) in the three comparisons (D3 vs. CK, D7 vs. CK, D3 vs. D7), 1,286 DEGs showed consistent upregulation or downregulation at D3 and D7 and were associated with pattern recognition, acute-phase response, complement cascade, chemokine and cytokine signaling, and apoptosis. The Real time quantitative PCR (RT-qPCR) analysis of eight DEGs confirmed the accuracy of the RNA-Sequencing (RNA-seq) data. Our results reflected a general process from pathogen recognition to inflammatory cytokine generation and delineated a putative Toll-like receptor signaling pathway in rainbow trout spleen, following F. psychrophilum infection. Taken together, these results provide new insights into the molecular mechanism of the immune response to F. psychrophilum infection and are a valuable resource for future research on the prevention and control of bacterial coldwater disease during salmon culture.
Dietary inosine monophosphate improved liver health and flesh quality of gibel carp (Carassius auratus gibelio) via activating AMPK signalling pathway and enhancing the contents of muscle fat and flavour substance
Authors: Cai Wanjie , Liu Haokun , Fu Lele , Han Dong , Zhu Xiaoming , Jin Junyan , Yang Yunxia , Xie Shouqi
Published: August 2022

Inosine monophosphate (IMP) improves the flesh quality of livestock and poultry, but studies about its effect on fishes are limited. The aim of the present study was to investigate the effect of exogenous IMP on the liver health and flesh quality of gibel carp (Carassius auratus gibelio). The results showed that IMP supplementation alleviated hepatic lipid deposition by increasing the AMP/ATP ratio and activating the AMPK signaling pathway then decreased inflammatory cytokine expression levels. Additionally, IMP increased the intramuscular fat content. Metabolomic analysis of meat revealed that the IMP, adenosine monophosphate (AMP), guanosine, inosine and purine metabolism pathways were involved in the improvements in fish meat quality, composition and taste that resulted from IMP supplementation. Our study provides a relatively comprehensive assessment of the effect of exogenous IMP on the liver metabolism and flesh quality of farmed fish. The results provide an effective food supplementation strategy for cultured fish.
Development of a novel score model to predict hyperinflammation in COVID-19 as a forecast of optimal steroid administration timing
Authors: Takeshita Yuichiro , Terada Jiro , Hirasawa Yasutaka , Kinoshita Taku , Tajima Hiroshi , Koshikawa Ken , Kinouchi Toru , Isaka Yuri , Shionoya Yu , Fujikawa Atsushi , Kato Yasuyuki , To Yasuo , Tada Yuji , Tsushima Kenji
Published: August 2022

Objectives

This study aims to create and validate a useful score system predicting the hyper-inflammatory conditions of COVID-19, by comparing it with the modified H-score.

Methods

A total of 98 patients with pneumonia (without oxygen therapy) who received initial administration of casirivimab/imdevimab or remdesivir were included in the study. The enrolled patients were divided into two groups: patients who required corticosteroid due to deterioration of pneumonia, assessed by chest X-ray or CT or respiratory failure, and those who did not, and clinical parameters were compared.

Results

Significant differences were detected in respiratory rate, breaths/min, SpO2, body temperature, AST, LDH, ferritin, and IFN-λ3 between the two groups. Based on the data, we created a corticosteroid requirement score: (1) the duration of symptom onset to treatment initiation ≥ 7 d, (2) the respiratory rate ≥ 22 breaths/min, (3) the SpO2 ≤ 95%, (4) BT ≥ 38.5°C, (5) AST levels ≥ 40 U/L, (6) LDH levels ≥ 340 U/L, (7) ferritin levels ≥ 800 ng/mL, and (8) IFN-λ3 levels ≥ 20 pg/mL. These were set as parameters of the steroid predicting score. Results showed that the area under the curve (AUC) of the steroid predicting score (AUC: 0.792, 95%CI: 0.698–0.886) was significantly higher than that of the modified H-score (AUC: 0.633, 95%CI: 0.502–0.764).

Conclusion

The steroid predicting score may be useful to predict the requirement of corticosteroid therapy in patients with COVID-19. The data may provide important information to facilitate a prospective study on a larger scale in this field.
Temporal variations of human and animal Rotavirus A genotypes in surface water used for drinking water production
Authors: Miura Takayuki , Kadoya Syun-suke , Takino Hiroyuki , Sano Daisuke , Akiba Michihiro
Published: August 2022

Rotavirus is a major cause of gastroenteritis among infants and children. In this study, nested PCR assays were developed to amplify partial regions of the VP7, VP4, and VP6 genes of Rotavirus A (RVA) for amplicon-based Illumina MiSeq sequencing to investigate RVA genotypes in environmental water samples. Eight sets of inner primers were first designed and screened for use in the nested PCR assays, and four sets of them could produce amplicons. Six sets of outer primers were then designed and combined with the four sets of inner primers that worked. The assays were evaluated for sensitivity using raw water samples collected from one drinking water treatment plant between April 2019 and March 2020 (Sample Set 1; N = 12) and seven DWTPs between 2018 and 2020 (Sample Set 2; N = 18). In total, 43 amplicons from Set 1 were sequenced and diverse sequences from human, porcine, bovine, equine, and feline RVA were observed. Human G8, G3, and G2 genotypes were obtained, with G8 predominant (relative abundance, 36–87%) in samples taken during the rotavirus epidemic season between April and June. Porcine G5, G11, and G4, and bovine G10 and G6 genotypes were also detected. VP4 sequence analysis revealed that the human P[8] genotype was present throughout the year, whereas P[4] and P[9] were present only in the epidemic season. The vaccine strains P[5] and P[8] (RotaTeq®) were also detected. Our approach enables the identification of prevalent human and animal RVA genotypes and their host species that potentially caused fecal contamination in water sources.
Deletion of the novel gene mother cell lysis X results in Cry1Ac encapsulation in the Bacillus thuringiensis HD73
Authors: Wang Jiaojiao , Yu Qingyue , Peng Qi , Slamti Leyla , Zhang Ruibin , Hou Shuo , Lereclus Didier , Song Fuping
Published: August 2022

The novel protein MclX (mother cell lysis X) in Bacillus thuringiensis subsp. kurstaki strain HD73 (B. thuringiensis HD73) was characterized in this work. MclX has no known domain and its gene deletion in HD73 resulted in Cry1Ac encapsulation in the mother cell and did not influence Cry1Ac protein production or insecticidal activity. In vitro cell wall hydrolysis experiments showed that MclX cannot hydrolyze the cell wall. In mclX deletion mutants, the expression of cwlC (which encodes a key cell wall hydrolase) was significantly decreased, as shown by the β-galactosidase activity assay. MclX cannot directly bind to the cwlC promoter, based on the electrophoretic mobility shift assay (EMSA). The cwlC was reported to be regulated by σK and GerE. However, the transcriptional activities of sigK and gerE showed no difference between HD73 and the mclX deletion mutant. It is indicated that MclX influenced cwlC expression independently of σK or GerE, through a new pathway to regulate cwlC expression. mclX deletion could be a new approach for insecticidal protein encapsulation in Bacillus thuringiensis.
Alleviating effects of gut micro-ecologically regulatory treatments on mice with constipation
Authors: Zhao Yueming , Liu Qingjing , Hou Yanmei , Zhao Yiqing
Published: August 2022

Treatments targeted for gut microbial regulation are newly developed strategies in constipation management. In this study, the alleviating effects of gut micro-ecologically regulatory treatments on constipation in mice were investigated. Male BALB/c mice were treated with loperamide to induce constipation, and then the corresponding intervention was administered in each group, respectively. The results showed that administration of mixed probiotics (MP), a 5-fold dose of postbiotics (P5), both synbiotics (S and S2), as well as mixed probiotics and postbiotics (MPP) blend for 8 days shortened the time to the first black stool, raised fecal water content, promoted intestinal motility, and increased serum motilin level in loperamide-treated mice. Furthermore, these treatments altered gut microbial composition and metabolism of short-chain fatty acids (SCFA). Based on linear regression analysis, SCFA was positively correlated with serum motilin except for isobutyrate. It suggested gut microbial metabolites affected secretion of motilin to increase gastrointestinal movement and transportation function and thus improved pathological symptoms of mice with constipation. In conclusion, the alteration of gut micro-ecology is closely associated with gastrointestinal function, and it is an effective way to improve constipation via probiotic, prebiotic, and postbiotic treatment.
The characteristics of protein-glutaminase from an isolated Chryseobacterium cucumeris strain and its deamidation application
Authors: Qu Ruidan , Dai Tian , Wu Jiajing , Tian Aitian , Zhang Yanfang , Kang Li , Ouyang Wei , Jin Congli , Niu Jinjin , Li Zhen , Chang Zhongyi , Jiang Deming , Huang Jing , Gao Hongliang
Published: August 2022

Protein-glutaminase (PG), a deamidation enzyme commercially derived from Chryseobacterium proteolyticum, is used to improve the solubility and other functional properties of food proteins. In this study, a new PG-producing strain, Chryseobacterium cucumeris ZYF120413-7, was isolated from soil, and it had a high PG yield and a short culture time. It gave the maximum PG activity with 0.557 U/ml on Cbz-Gln-Gly after 12 h of culture, indicating that it was more suitable for PG production. The enzyme activity recovery and purification fold were 32.95% and 161.95-fold, respectively, with a specific activity of 27.37 U/mg. The PG was a pre-pro-protein with a 16 amino acids putative signal peptide, a pro-PG of 118 amino acids, and a mature PG of 185 amino acids. The amino acid sequence identity of PG from strain ZYF120413-7 was 74 and 45%, respectively, to that of PG from C. proteolyticum 9670T and BH-PG. The optimum reaction pH and temperature of PG was 6 and 60°C, respectively. Enzyme activity was inhibited by Cu2+. The optimum PG substrate was Cbz-Gln-Gly, and the Km and Vmax values were 1.68 mM and 1.41 μM mg protein−1 min−1, respectively. Degree of deamidation (DD) of soy protein isolate (SPI) treated by purified PG was 40.75% within the first 2 h and 52.35% after 18 h. These results demonstrated that the PG from C. cucumeris ZYF120413-7 was a promising protein-deamidating enzyme for improving the functionality of food proteins.