bluuebunny/crossref_metadata_2025 huggingface dataset:
Establishment of Novel Genotoxicity Assay System Using Murine Normal Epithelial Tissue-Derived Organoids
Authors: Komiya Masami , Ishigamori Rikako , Naruse Mie , Ochiai Masako , Miyoshi Noriyuki , Imai Toshio , Totsuka Yukari
Published: November 2021

Short-/middle-term and simple prediction studies for carcinogenesis are needed for the safety assessment of chemical substances. To establish a novel genotoxicity assay with an in vivo mimicking system, we prepared murine colonic/pulmonary organoids from gpt delta mice according to the general procedure using collagenase/dispase and cultured them in a 3D environment. When the organoids were exposed to foodborne carcinogens—2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine (PhIP) and acrylamide (AA)—in the presence of metabolic activation systems, mutation frequencies (MFs) occurring in the gpt gene dose-dependently increased. Moreover, the mutation spectrum analysis indicated predominant G:C to T:A transversion with PhIP, and A:T to C:G and A:T to T:A transversion with AA. These data correspond to those of a previous study describing in vivo mutagenicity in gpt delta mice. However, organoids derived from the liver, a non-target tissue of PhIP-carcinogenesis, also demonstrated genotoxicity with a potency comparable to colonic organoids. Organoids and PhIP were directly incubated in the presence of metabolic activation systems; therefore, there was a lack of organ specificity, as observed in vivo. Additionally, PhIP-DNA adduct levels were comparable in hepatic and colonic organoids after PhIP exposure. Taken together, the organoids prepared in the present study may be helpful to predict chemical carcinogenesis.
Transcriptome Profiling of the Salt Stress Response in the Leaves and Roots of Halophytic Eutrema salsugineum
Authors: Li Chuanshun , Qi Yuting , Zhao Chuanzhi , Wang Xingjun , Zhang Quan
Published: November 2021

Eutrema salsugineum can grow in natural harsh environments; however, the underlying mechanisms for salt tolerance of Eutrema need to be further understood. Herein, the transcriptome profiling of Eutrema leaves and roots exposed to 300 mM NaCl is investigated, and the result emphasized the role of genes involved in lignin biosynthesis, autophagy, peroxisome, and sugar metabolism upon salt stress. Furthermore, the expression of the lignin biosynthesis and autophagy-related genes, as well as 16 random selected genes, was validated by qRT-PCR. Notably, the transcript abundance of a large number of lignin biosynthesis genes such as CCoAOMT, C4H, CCR, CAD, POD, and C3′H in leaves was markedly elevated by salt shock. And the examined lignin content in leaves and roots demonstrated salt stress led to lignin accumulation, which indicated the enhanced lignin level could be an important mechanism for Eutrema responding to salt stress. Additionally, the differentially expressed genes (DEGs) assigned in the autophagy pathway including Vac8, Atg8, and Atg4, as well as DEGs enriched in the peroxisome pathway such as EsPEX7, EsCAT, and EsSOD2, were markedly induced in leaves and/or roots. In sugar metabolism pathways, the transcript levels of most DEGs associated with the synthesis of sucrose, trehalose, raffinose, and xylose were significantly enhanced. Furthermore, the expression of various stress-related transcription factor genes including WRKY, AP2/ERF-ERF, NAC, bZIP, MYB, C2H2, and HSF was strikingly improved. Collectively, the increased expression of biosynthesis genes of lignin and soluble sugars, as well as the genes in the autophagy and peroxisome pathways, suggested that Eutrema encountering salt shock possibly possess a higher capacity to adjust osmotically and facilitate water transport and scavenge reactive oxidative species and oxidative proteins to cope with the salt environment. Thus, this study provides a new insight for exploring the salt tolerance mechanism of halophytic Eutrema and discovering new gene targets for the genetic improvement of crops.
LIMD2 is a Prognostic and Predictive Marker in Patients With Esophageal Cancer Based on a ceRNA Network Analysis
Authors: Chen Yuanmei , Huang Xinyi , Zhu Kunshou , Li Changkun , Peng Haiyan , Chen Lin , Huang Zhengrong , Zhang Yangfan , Weng Guibin , Xiao Tianya , Chen Junqiang , Xu Yuanji
Published: November 2021

Globally, esophageal cancer (ECA) is the seventh most common cancer and sixth most common cause of cancer-associated mortality. However, there are no reliable prognostic and predictive molecular markers for ECA; in addition, the pathogenesis of ECA is not fully elucidated. The expressions of circular RNAs (circRNAs), microRNAs (miRNAs), and messenger RNAs (mRNAs) of ECA and control groups were obtained from the RNA-sequencing (RNA-seq) data of our hospital, the Gene Expression Omnibus (GEO), and The Cancer Genome Atlas (TCGA) datasets. Analyses of differentially expressed genes, the circRNA–miRNA–mRNA–competing endogenous RNA (ceRNA) network, and functional/pathway enrichment were conducted. The key targets in the ceRNA network that showed significant results in survival Cox regression analyses were selected. Furthermore, analyses of immune infiltration and autophagy genes related to the key targets were performed. Seven circRNAs, 22 miRNAs, and 34 mRNAs were identified as vital genes in ECA; the nuclear factor-κ-gene binding (NF-κB) and phosphatidylinositol-3 kinase/protein kinase B (PI3K-Akt) signaling were identified as the most enriched pathways. In addition, the LIM domain containing 2 (LIMD2) was an independent predictor of prognosis in ECA patients and closely associated with immunity and autophagy. Moreover, quantitative reverse-transcription polymerase chain reaction (qRT-PCR) revealed significant upregulation of LIMD2 expression in ECA tissues. ECA may be closely correlated with NF-κB and PI3K/Akt signaling. In addition, LIMD2 could be a potential prognostic and predictive marker of ECA.
Modulation of Cystatin C in Human Macrophages Improves Anti-Mycobacterial Immune Responses to Mycobacterium tuberculosis Infection and Coinfection With HIV
Authors: Pires David , Calado Marta , Velez Tomás , Mandal Manoj , Catalão Maria João , Neyrolles Olivier , Lugo-Villarino Geanncarlo , Vérollet Christel , Azevedo-Pereira José Miguel , Anes Elsa
Published: November 2021

Tuberculosis owes its resurgence as a major global health threat mostly to the emergence of drug resistance and coinfection with HIV. The synergy between HIV andMycobacterium tuberculosis(Mtb) modifies the host immune environment to enhance both viral and bacterial replication and spread. In the lung immune context, both pathogens infect macrophages, establishing favorable intracellular niches. Both manipulate the endocytic pathway in order to avoid destruction. Relevant players of the endocytic pathway to control pathogens include endolysosomal proteases, cathepsins, and their natural inhibitors, cystatins. Here, a mapping of the human macrophage transcriptome for type I and II cystatins during Mtb, HIV, or Mtb-HIV infection displayed different profiles of gene expression, revealing cystatin C as a potential target to control mycobacterial infection as well as HIV coinfection. We found that cystatin C silencing in macrophages significantly improves the intracellular killing of Mtb, which was concomitant with an increased general proteolytic activity of cathepsins. In addition, downmodulation of cystatin C led to an improved expression of the human leukocyte antigen (HLA) class II in macrophages and an increased CD4+T-lymphocyte proliferation along with enhanced IFN-γ secretion. Overall, our results suggest that the targeting of cystatin C in human macrophages represents a promising approach to improve the control of mycobacterial infections including multidrug-resistant (MDR) TB.
The Bacterial Mucosal Immunotherapy MV130 Protects Against SARS-CoV-2 Infection and Improves COVID-19 Vaccines Immunogenicity
Authors: del Fresno Carlos , García-Arriaza Juan , Martínez-Cano Sarai , Heras-Murillo Ignacio , Jarit-Cabanillas Aitor , Amores-Iniesta Joaquín , Brandi Paola , Dunphy Gillian , Suay-Corredera Carmen , Pricolo Maria Rosaria , Vicente Natalia , López-Perrote Andrés , Cabezudo Sofía , González-Corpas Ana , Llorca Oscar , Alegre-Cebollada Jorge , Garaigorta Urtzi , Gastaminza Pablo , Esteban Mariano , Sancho David
Published: November 2021

COVID-19-specific vaccines are efficient prophylactic weapons against SARS-CoV-2 virus. However, boosting innate responses may represent an innovative way to immediately fight future emerging viral infections or boost vaccines. MV130 is a mucosal immunotherapy, based on a mixture of whole heat-inactivated bacteria, that has shown clinical efficacy against recurrent viral respiratory infections. Herein, we show that the prophylactic intranasal administration of this immunotherapy confers heterologous protection against SARS-CoV-2 infection in susceptible K18-hACE2 mice. Furthermore, in C57BL/6 mice, prophylactic administration of MV130 improves the immunogenicity of two different COVID-19 vaccine formulations targeting the SARS-CoV-2 spike (S) protein, inoculated either intramuscularly or intranasally. Independently of the vaccine candidate and vaccination route used, intranasal prophylaxis with MV130 boosted S-specific responses, including CD8+-T cell activation and the production of S-specific mucosal IgA antibodies. Therefore, the bacterial mucosal immunotherapy MV130 protects against SARS-CoV-2 infection and improves COVID-19 vaccines immunogenicity.
The Alternatively Spliced Isoforms of Key Molecules in the cGAS-STING Signaling Pathway
Authors: Liang Jiaqian , Hong Ze , Sun Boyue , Guo Zhaoxi , Wang Chen , Zhu Juanjuan
Published: November 2021

Alternative splicing of pre-mRNA increases transcriptome and proteome diversity by generating distinct isoforms that encode functionally diverse proteins, thus affecting many biological processes, including innate immunity. cGAS-STING signaling pathway, whose key molecules also undergo alternative splicing, plays a crucial role in regulating innate immunity. Protein isoforms of key components in the cGAS-STING-TBK1-IRF3 axis have been detected in a variety of species. A chain of evidence showed that these protein isoforms exhibit distinct functions compared to their normal counterparts. The mentioned isoforms act as positive or negative modulators in interferon response via distinct mechanisms. Particularly, we highlight that alternative splicing serves a vital function for the host to avoid the overactivation of the cGAS-STING signaling pathway and that viruses can utilize alternative splicing to resist antiviral response by the host. These findings could provide insights for potential alternative splicing-targeting therapeutic applications.
O-GlcNAcylation in Chronic Lymphocytic Leukemia and Other Blood Cancers
Authors: Spaner David E.
Published: November 2021

In the past decade, aberrant O-GlcNAcylation has emerged as a new hallmark of cancer. O-GlcNAcylation is a post-translational modification that results when the amino-sugar β-D-N-acetylglucosamine (GlcNAc) is made in the hexosamine biosynthesis pathway (HBP) and covalently attached to serine and threonine residues in intracellular proteins by the glycosyltransferase O-GlcNAc transferase (OGT). O-GlcNAc moieties reflect the metabolic state of a cell and are removed by O-GlcNAcase (OGA). O-GlcNAcylation affects signaling pathways and protein expression by cross-talk with kinases and proteasomes and changes gene expression by altering protein interactions, localization, and complex formation. The HBP and O-GlcNAcylation are also recognized to mediate survival of cells in harsh conditions. Consequently, O-GlcNAcylation can affect many of the cellular processes that are relevant for cancer and is generally thought to promote tumor growth, disease progression, and immune escape. However, recent studies suggest a more nuanced view with O-GlcNAcylation acting as a tumor promoter or suppressor depending on the stage of disease or the genetic abnormalities, proliferative status, and state of the p53 axis in the cancer cell. Clinically relevant HBP and OGA inhibitors are already available and OGT inhibitors are in development to modulate O-GlcNAcylation as a potentially novel cancer treatment. Here recent studies that implicate O-GlcNAcylation in oncogenic properties of blood cancers are reviewed, focusing on chronic lymphocytic leukemia and effects on signal transduction and stress resistance in the cancer microenvironment. Therapeutic strategies for targeting the HBP and O-GlcNAcylation are also discussed.
Conserved Induction of Distinct Antiviral Signalling Kinetics by Primate Interferon Lambda 4 Proteins
Authors: Guo Cuncai , Reuss Dorothee , Coey Jonathon D. , Sukumar Swathi , Lang Benjamin , McLauchlan John , Boulant Steeve , Stanifer Megan L. , Bamford Connor G. G.
Published: November 2021

Interferon lambdas (IFNλ) (also known as type III IFNs) are critical cytokines that combat infection predominantly at barrier tissues, such as the lung, liver, and gastrointestinal tract. Humans have four IFNλs (1–4), where IFNλ1–3 show ~80%–95% homology, and IFNλ4 is the most divergent displaying only ~30% sequence identity. Variants in IFNλ4 in humans are associated with the outcome of infection, such as with hepatitis C virus. However, how IFNλ4 variants impact cytokine signalling in other tissues and how well this is conserved is largely unknown. In this study, we address whether differences in antiviral signalling exist between IFNλ4 variants in human hepatocyte and intestinal cells, comparing them to IFNλ3. We demonstrate that compared to IFNλ3, wild-type human IFNλ4 induces a signalling response with distinct magnitudes and kinetics, which is modified by naturally occurring variants P70S and K154E in both cell types. IFNλ4’s distinct antiviral response was more rapid yet transient compared to IFNλ1 and 3. Additionally, divergent antiviral kinetics were also observed using non-human primate IFNλs and cell lines. Furthermore, an IFNλ4-like receptor-interacting interface failed to alter IFNλ1’s kinetics. Together, our data provide further evidence that major functional differences exist within the IFNλ gene family. These results highlight the possible tissue specialisation of IFNλs and encourage further investigation of the divergent, non-redundant activities of IFNλ4 and other IFNλs.
Case Report: Infantile Urticaria as a Herald of Neonatal Onset Multisystem Inflammatory Disease With a Novel Mutation in NLRP3
Authors: Patrick Anna E. , Lyons Eden M. , Ishii Lisa , Boyd Alan S. , Choi Joseph M. , Dewan Anna K. , Markle Janet G.
Published: November 2021

Neonatal multisystem onset inflammatory disorder (NOMID) is a severe autoinflammatory syndrome that can have an initial presentation as infantile urticaria. Thus, an immediate recognition of the clinical symptoms is essential for obtaining a genetic diagnosis and initiation of early therapies to prevent morbidity and mortality. Herein, we describe a neonate presenting with urticaria and systemic inflammation within hours after birth who developed arthropathy and neurologic findings. Pathologic evaluation of the skin revealed an infiltration of lymphocytes, eosinophils, and scattered neutrophils. Genetic analysis identified a novel heterozygous germline variant of unknown significance in the NLRP3 gene, causing the missense mutation M408T. Variants of unknown significance are common in genetic sequencing studies and are diagnostically challenging. Functional studies of the M408T variant demonstrated enhanced formation and activity of the NLRP3 inflammasome, with increased cleavage of the inflammatory cytokine IL-1β. Upon initiation of IL-1 pathway blockade, the infant had a robust response and improvement in clinical and laboratory findings. Our experimental data support that this novel variant in NLRP3 is causal for this infant’s diagnosis of NOMID. Rapid assessment of infantile urticaria with biopsy and genetic diagnosis led to early recognition and targeted anti-cytokine therapy. This observation expands the NOMID-causing variants in NLRP3 and underscores the role of genetic sequencing in rapidly identifying and treating autoinflammatory disease in infants. In addition, these findings highlight the importance of establishing the functional impact of variants of unknown significance, and the impact this knowledge may have on therapeutic decision making.
Adoptive Transfer of Immune Cells Into RAG2IL-2Rγ-Deficient Mice During Litomosoides sigmodontis Infection: A Novel Approach to Investigate Filarial-Specific Immune Responses
Authors: Wiszniewsky Anna , Layland Laura E. , Arndts Kathrin , Wadephul Lisa M. , Tamadaho Ruth S. E. , Borrero-Wolff Dennis , Chunda Valerine C. , Kien Chi Anizette , Hoerauf Achim , Wanji Samuel , Ritter Manuel
Published: November 2021

Despite long-term mass drug administration programmes, approximately 220 million people are still infected with filariae in endemic regions. Several research studies have characterized host immune responses but a major obstacle for research on human filariae has been the inability to obtain adult worms which in turn has hindered analysis on infection kinetics and immune signalling. Although theLitomosoides sigmodontisfilarial mouse model is well-established, the complex immunological mechanisms associated with filarial control and disease progression remain unclear and translation to human infections is difficult, especially since human filarial infections in rodents are limited. To overcome these obstacles, we performed adoptive immune cell transfer experiments into RAG2IL-2Rγ-deficient C57BL/6 mice. These mice lack T, B and natural killer cells and are susceptible to infection with the human filariaLoa loa. In this study, we revealed a long-term release ofL. sigmodontisoffspring (microfilariae) in RAG2IL-2Rγ-deficient C57BL/6 mice, which contrasts to C57BL/6 mice which normally eliminate the parasites before patency. We further showed that CD4+T cells isolated from acuteL. sigmodontis-infected C57BL/6 donor mice or mice that already cleared the infection were able to eliminate the parasite and prevent inflammation at the site of infection. In addition, the clearance of the parasites was associated with Th17 polarization of the CD4+T cells. Consequently, adoptive transfer of immune cell subsets into RAG2IL-2Rγ-deficient C57BL/6 mice will provide an optimal platform to decipher characteristics of distinct immune cells that are crucial for the immunity against rodent and human filarial infections and moreover, might be useful for preclinical research, especially about the efficacy of macrofilaricidal drugs.
MG1141A as a Highly Potent Monoclonal Neutralizing Antibody Against SARS-CoV-2 Variants
Authors: Lee Sua , Jang Shina , Kang Jihoon , Park Soo Bin , Han Young Woo , Nam Hyemi , Kim Munkyung , Lee Jeewon , Cho Ki Joon , Kim Jeonghun , Oh Miyoung , Ryu Jihye , Seok Jong Hyeon , Kim Yunhwa , Lee Jee-Boong , Park Man-Seong , Kim Yong-Sung , Park Hosun , Kim Dong-Sik
Published: November 2021

Since the coronavirus disease outbreak in 2019, several antibody therapeutics have been developed to treat severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections. Antibody therapeutics are effective in neutralizing the virus and reducing hospitalization in patients with mild and moderate infections. These therapeutics target the spike protein of SARS-CoV-2; however, emerging mutations in this protein reduce their efficiency. In this study, we developed a universal SARS-CoV-2 neutralizing antibody. We generated a humanized monoclonal antibody, MG1141A, against the receptor-binding domain of the spike protein through traditional mouse immunization. We confirmed that MG1141A could effectively neutralize live viruses, with an EC50 of 92 pM, and that it exhibited effective Fc-mediated functions. Additionally, it retained its neutralizing activity against the alpha (UK), beta (South Africa), and gamma (Brazil) variants of SARS-CoV-2. Taken together, our study contributes to the development of a novel antibody therapeutic approach, which can effectively combat emerging SARS-CoV-2 mutations.
A Model of Minor Histocompatibility Antigens in Allogeneic Hematopoietic Cell Transplantation
Authors: Martin Paul J. , Levine David M. , Storer Barry E. , Zheng Xiuwen , Jain Deepti , Heavner Ben , Norris Brandon M. , Geraghty Daniel E. , Spellman Stephen R. , Sather Cassie L. , Wu Feinan , Hansen John A.
Published: November 2021

Minor histocompatibility antigens (mHAg) composed of peptides presented by HLA molecules can cause immune responses involved in graft-versus-host disease (GVHD) and graft-versus-leukemia effects after allogeneic hematopoietic cell transplantation (HCT). The current study was designed to identify individual graft-versus-host genomic mismatches associated with altered risks of acute or chronic GVHD or relapse after HCT between HLA-genotypically identical siblings. Our results demonstrate that in allogeneic HCT between a pair of HLA-identical siblings, a mHAg manifests as a set of peptides originating from annotated proteins and non-annotated open reading frames, which i) are encoded by a group of highly associated recipient genomic mismatches, ii) bind to HLA allotypes in the recipient, and iii) evoke a donor immune response. Attribution of the immune response and consequent clinical outcomes to individual peptide components within this set will likely differ from patient to patient according to their HLA types.
T-Cell Epitopes Shared Between Immunizing HLA and Donor HLA Associate With Graft Failure After Kidney Transplantation
Authors: Peereboom Emma T. M. , Matern Benedict M. , Tomosugi Toshihide , Niemann Matthias , Drylewicz Julia , Joosten Irma , Allebes Wil A. , van der Meer Arnold , Hilbrands Luuk B. , Baas Marije C. , van Reekum Franka E. , Verhaar Marianne C. , Kamburova Elena G. , Seelen Marc A. J. , Sanders Jan Stephan , Hepkema Bouke G. , Lambeck Annechien J. , Bungener Laura B. , Roozendaal Caroline , Tilanus Marcel G. J. , Voorter Christien E. , Wieten Lotte , van Duijnhoven Elly M. , Gelens Mariëlle A. C. J. , Christiaans Maarten H. L. , van Ittersum Frans J. , Nurmohamed Azam , Lardy Neubury M. , Swelsen Wendy , van der Pant Karlijn A. , van der Weerd Neelke C. , ten Berge Ineke J. M. , Bemelman Fréderike J. , de Vries Aiko P. J. , de Fijter Johan W. , Betjes Michiel G. H. , Roelen Dave L. , Claas Frans H. , Otten Henny G. , Heidt Sebastiaan , van Zuilen Arjan D. , Kobayashi Takaaki , Geneugelijk Kirsten , Spierings Eric
Published: November 2021

CD4+T-helper cells play an important role in alloimmune reactions following transplantation by stimulating humoral as well as cellular responses, which might lead to failure of the allograft. CD4+memory T-helper cells from a previous immunizing event can potentially be reactivated by exposure to HLA mismatches that share T-cell epitopes with the initial immunizing HLA. Consequently, reactivity of CD4+memory T-helper cells toward T-cell epitopes that are shared between immunizing HLA and donor HLA could increase the risk of alloimmunity following transplantation, thus affecting transplant outcome. In this study, the amount of T-cell epitopes shared between immunizing and donor HLA was used as a surrogate marker to evaluate the effect of donor-reactive CD4+memory T-helper cells on the 10-year risk of death-censored kidney graft failure in 190 donor/recipient combinations using the PIRCHE-II algorithm. The T-cell epitopes of the initial theoretical immunizing HLA and the donor HLA were estimated and the number of shared PIRCHE-II epitopes was calculated. We show that the natural logarithm-transformed PIRCHE-II overlap score, or Shared T-cell EPitopes (STEP) score, significantly associates with the 10-year risk of death-censored kidney graft failure, suggesting that the presence of pre-transplant donor-reactive CD4+memory T-helper cells might be a strong indicator for the risk of graft failure following kidney transplantation.
Shifts in Estuarine Macroinvertebrate Communities Associated With Water Quality and Climate Change
Authors: Goulding Thomas , Sousa Pedro M. , Silva Gilda , Medeiros João Paulo , Carvalho Frederico , Metelo Inês , Freitas Catarina , Lopes Nuno , Chainho Paula , Costa José Lino
Published: November 2021

The present work aims to identify changes in the macroinvertebrate community of the Tagus estuary (Portugal) due to improvements in water quality and to climate change. Data was collected over a period of 16 years (1998–2014) from different sites located along the estuarine gradient. The AZTI Marine Biotic Index (AMBI) was used to assess the ecological quality status based on benthic invertebrate communities and identify possible variations associated with changes in water quality. The overall distribution of each species was examined to detect possible changes associated with climate, based on species’ affinity for more temperate or subtropical climates. Results demonstrate that there was an overall improvement of AMBI scores during the assessment period. The analysis of the geographical distribution of benthic species seems to indicate that there has been an increase of species which prefer subtropical climates in the shallower waters of the estuary, whereas in the deeper estuarine sections the propensity is for species that prefer temperate climates.
Genetically Engineering Coral for Conservation: Psychological Correlates of Public Acceptability
Authors: Mankad Aditi , Hobman Elizabeth V. , Carter Lucy
Published: November 2021

Coral bleaching contributes to widespread reef loss globally, including Australia’s World Heritage site, the Great Barrier Reef. Synthetic biology offers the potential to isolate and cultivate strains of coral that can naturally withstand higher sea surface temperatures associated with climate change. A national survey was conducted (N = 1,148 Australians) measuring psychological predictors of support for a synthetic biology conservation solution to coral loss. The analysis showed a partially mediated path model was useful in explaining a significant amount of variance in public support for the development of genetically engineered coral for conservation (R2 = 0.40) and in willingness to visit parts of the Great Barrier Reef where genetically engineered coral had (hypothetically) been introduced (R2 = 0.24). Participants were moderately strongly supportive of technology development and were most keen to implement genetically engineered coral with between 50 and 70% of reef remaining intact; recent estimates of coral cover across the Great Barrier Reef are well below that already. There was a negative association between perceived risks of genetically engineered coral and public support; however, perceived benefit of genetically engineered coral in protecting the reef and relative advantage of a synthetic biology solution over existing protection strategies were the most influential predictors of public support. The findings suggest that the general public are not averse to the development of a synthetic biology solution for restoring the reef, and they may be especially influenced by whether the synthetic biology solution is shown to be efficacious, particularly in comparison to other conservation solutions. However, support for a synthetic biology intervention is conditional and many participants expressed concerns about possible long-term impacts on humans, animals, and the environment as a result of deploying engineered coral.
Exact Zoning Optimization Model for Marine Spatial Planning (MSP)
Authors: Basirati Mohadese , Billot Romain , Meyer Patrick , Bocher Erwan
Published: November 2021

Marine spatial planning (MSP) has recently attracted more attention as an efficient decision support tool. MSP is a strategic and long-term process gathering multiple competing users of the ocean with the objective to simplify decisions regarding the sustainable use of marine resources. One of the challenges in MSP is to determine an optimal zone to locate a new activity while taking into account the locations of the other existing activities. Most approaches to spatial zoning are formulated as non-linear optimization models involving multiple objectives, which are usually solved using stochastic search algorithms, leading to sub-optimal solutions. In this paper, we propose to model the problem as a Multi-Objective Integer Linear Program. The model is developed for raster data and it aims at maximizing the interest of the area of the zone dedicated to the new activity while maximizing its spatial compactness. We study two resolution methods: first, a weighted-sum of the two objectives, and second, an interactive approach based on an improved augmented version of the ϵ-constraint method, AUGMECON2. To validate and study the model, we perform experiments on artificially generated data. Our experimental study shows that AUGMECON2 represents the most promising approach in terms of relevance and diversity of the solutions, compactness, and computation time.
Corrosion Initiation Behavior of Thermal Control Oxidation Film of LA103Z Alloy
Authors: Wen Chen , Bai Jingying , Zhao Kuo , Cui Qingxin , Jiao Baocheng , Wang Xuguang , Zhang Ligong
Published: November 2021

The magnesium lithium thermal control oxidation is a commonly used conductive anti-corrosion treatment method for aerospace. The corrosion behaviors of thermal control oxidation films for LA103Z alloys were studied in 3.5% NaCl solution. The corrosion characteristics with different immersion time were characterized by using scanning electron microscopy, energy spectroscopy, and electrochemical methods. The results showed that the corrosion of Mg-Li alloy with chemical oxidation film starts from pitting corrosion, gradually expands in depth in the early stage, forms corrosion holes, and then gradually develops into river-like morphology. In the last stage, the increasing corrosion products slow down the corrosion rate and gradually covers the entire sample surface. The corrosion product mainly consists of MgO, LiF, and MgCl2. Combined with the electrochemical characteristics, the corrosion mechanism was investigated.
Severe Adverse Toxic Effects of Low-Dose Methotrexate Treatment on an Ectopic Pregnancy Patient With Methylenetetrahydrofolate Reductase Mutations: A Case Report
Authors: Yu Huan , Wang Wenhui , Liang Haiyan , Wang Kun , Ling Bin
Published: November 2021

Background: Low-dose methylenetetrahydrofolate (LD-MTX) has been widely used for the treatment of the ectopic pregnancy (EP) for many decades, and related severe adverse toxic effects are rare. Current studies have shown that the polymorphisms of methylenetetrahydrofolate reductase (MTHFR) gene can decrease the MTX clearance, leading to the metabolite accumulation. However, there is a lack of literature report on an MTHFR gene polymorphism associated with adverse toxic effects resulting from the use of LD-MTX in an EP.

Case Presentation: We report a rare case of a 38-year-old female who developed persistent fever, grade IV myelosuppression, skin lesions, mucositis, and liver injury after single dose of LDMTX to treat EP. The personalized genetic testing showed that MTHFR TT (677C>T) and MTHFR AA (1298A>C) were detected. Gradually, the symptoms improved after calcium leucovorin (CF) rescue, continuous renal replacement therapy (CRRT), promoting blood system regeneration, and multiple supportive treatments.

Conclusion: This is the first report on the serious adverse toxic effects of LD-MTX on an EP patient with MTHFR mutations. We aim to alert obstetricians and gynecologists to this rare condition. The unexpected life-threatening toxicity with LD-MTX should be highly considered and recognized early. In particular, some easily overlooked gastrointestinal, skin, and mucosal symptoms occur earlier than severe myelosuppression. When toxic effects are suspected, detecting the polymorphisms of an MTHFR gene and monitoring MTX concentration in blood could assist us to formulate individualized and active treatments.
Passive Surveillance of Malaria in Pregnant Women, Non-pregnant Women and Children Under 5 Years of Age in Bannu District, Khyber Pakhtunkhwa Pakistan
Authors: Qureshi Humera , Khan Muhammad Imran , Ahmad Akhlaq , Awan Usman Ayub , Khattak Aamer Ali , Khan Ayesha Ali , Sun Yaqiong
Published: November 2021

Background: Malaria among pregnant women is one of the major causes of maternal and infant mortality and morbidity, especially in high-risk areas. Therefore, our study identified the burden of malaria for pregnant women, non-pregnant women, and children under 5 years of age, and malaria service health facilities in Bannu district, Khyber Pakhtunkhwa, Pakistan.

Methods: A cross-sectional study was conducted. In this survey, 15,650 individuals were surveyed, and 1,283 were malaria-positive detected. The data were collected from 80 different healthcare centers. SPSS version 23 was used for data analysis. ArcGIS version 10.8 was used for study area mapping.

Results: Malaria was detected in 23.3% of children under five, 4.4% of pregnant women, and 72.3% of non-pregnant women, respectively. Moreover, P. falciparum, P. vivax, and mixed infection had a prevalence of 2.1, 96.8, and 1.1%. The most often used and effective medications to treat malaria were chloroquine (29.7%) and primaquine (69.4%).

Conclusion: This study's findings depict that malaria's prevalence in the non-pregnant women's group was high. Additionally, P. vivax infection was found to be more prevalent than other types of malaria infection. Due to the scarcity of healthcare facilities in this endemic region, special attention should be directed to strengthening the malaria surveillance and eradication programs.
Increasing Burden of Nursing Care on the Treatment of COVID-19 Patients in the Aging Society: Analyses During the First to the Third Wave of Pandemic in Kyoto City, Japan
Authors: Fujita Kohei , Kashihara Eriko , Kanai Osamu , Hata Hiroaki , Yasoda Akihiro , Odagaki Takao , Mio Tadashi
Published: November 2021

Background: The coronavirus disease 2019 (COVID-19) pandemic is associated with a heavy burden on patient's mental and physical health, regional healthcare resources, and global economic activity. An aging society such as Japan has many retirement homes and long-term stay hospitals for the elderly and their inhabitants. During the COVID-19 pandemic, disease clusters are often identified in retirement homes and long-term stay hospitals. Although we hypothesize that additional burdens of nursing care for elderly patients will reinforce the anxiety and exhaustion of medical staff and healthcare resources in the aging society, the actual situation is not well understood. In this study, we aimed to evaluate the current situation and countermeasures of the COVID-19 pandemic in the aging society.

Methods: We reviewed COVID-19 patients who required hospitalization at the National Hospital Organization Kyoto Medical Center, a 600-bed capacity hospital located in Kyoto, Japan, between 1 April 2020 and 31 March 2021. We assessed the characteristics of the COVID-19 patients, disease severity, duration of hospitalization, outcome at discharge, degree of activities of daily living (ADLs), and complications unique to elderly patients.

Results: We enrolled 118 patients who required hospitalization during the study period. Approximately 40% of the patients were aged ≥ 80 years. Dementia (27.1%) was the most prevalent underlying disease, followed by diabetes mellitus (23.7%) and chronic kidney disease (23.7%). Approximately 60% of hospitalized COVID-19 patients had impaired ADL at admission. The COVID-19 patients aged 80 years or older required significantly more longer-term hospitalization than the COVID-19 patients aged under 80 years (15.5 ± 8.2 vs. 13.1 ± 7.7, P = 0.032). In elderly patients aged 80 years or older, approximately 50% of patients had geriatric mental disorders, and approximately 70% had bedridden status and feeding difficulty. Poor ADL at admission was significantly associated with COVID-19 mortality (Odds ratio, 5.6; 95% confidence interval, 1.04–45.2; p-value = 0.044).

Conclusions: The proportion of elderly patients aged 80 years or older was relatively high during the hospitalization for COVID-19. Poor ADL at admission in these elderly patients was significantly associated with poor prognosis of COVID-19. We should keep in mind that healthcare workers are forced to have an additional burden of nursing care in the aging society during the COVID-19 pandemic. Therefore, interventions to reduce the burden are urgently required.
Treatment of Dry Eye With Intracanalicular Injection of Hydroxybutyl Chitosan: A Prospective Randomized Clinical Trial
Authors: Lin Tong , Wang Wushuang , Lu Yang , Gong Lan
Published: November 2021

Background: Punctal/intracanalicular plugs on the market nowadays are all designed before clinical use in treating dry eye disease (DED). To provide an individualized lacrimal drainage system occlusion method and reduce the complications, we developed a “liquid plug” strategy by intracanalicular injection of hydroxybutyl chitosan (HBC) solution, a thermosensitive, phase-changing biomaterial. This study evaluated the efficacy and safety of the HBC plug in treating dry eye disease by comparing it with the VisiPlug absorbable intracanalicular plug.

Methods: A monocenter, randomized, controlled clinical trial was performed. Fifty patients with DED were randomized 1:1 to undergo either the HBC injection treatment or the VisiPlug treatment. Ocular Surface Disease Index (OSDI) questionnaire, tear break-up time (TBUT), corneal fluorescence staining (CFS), tear meniscus height (TMH), and phenol red thread test were evaluated at Day 0 (baseline, before treatment) and Weeks 1, 4, and 12.

Results: The two groups had a balanced baseline of age, gender, and DED-related characteristics. Both occlusion methods could relieve the symptoms and signs of DED. Significant improvement was found in OSDI, phenol red thread test, and tear meniscus height (P < 0.05 compared to baseline) but not in corneal fluorescence staining and tear break-up time (P > 0.05). There is no statistically significant difference between HBC injection and VisiPlug at Weeks 1 and 4 (P > 0.05). However, at week 12, the HBC injection was not as effective as the VisiPlug in maintaining phenol red thread test (HBC: 5.35 ± 3.22 mm, VisiPlug: 8.59 ± 4.35 mm, P = 0.009) and tear meniscus height (HBC: 206.9 ± 47.95 μm, VisiPlug: 242.59 ± 60.30 μm, P = 0.041). The numbers of ocular adverse events were relatively low in both groups.

Conclusions: The HBC injection showed similar efficacy and safety compared to VisiPlug. The intracanalicular injection of HBC solution proves to be a promising, individualizing method to treat DED.

Clinical Trial Registration: This study is registered with the Chinese Clinical Trial Registry (https://www.chictr.org.cn/enindex.aspx), Identifier: ChiCTR1800016603.
Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern
Authors: Alves Pedro A. , de Oliveira Ellen G. , Franco-Luiz Ana Paula M. , Almeida Letícia T. , Gonçalves Amanda B. , Borges Iara A. , Rocha Flávia de S. , Rocha Raissa P. , Bezerra Matheus F. , Miranda Pâmella , Capanema Flávio D. , Martins Henrique R. , Weber Gerald , Teixeira Santuza M. R. , Wallau Gabriel Luz , do Monte-Neto Rubens L.
Published: November 2021

The coronavirus disease 2019 (COVID-19) pandemic unfolded due to the widespread severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission reinforced the urgent need for affordable molecular diagnostic alternative methods for massive testing screening. We present the clinical validation of a pH-dependent colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) for SARS-CoV-2 detection. The method revealed a limit of detection of 19.3 ± 2.7 viral genomic copies/μL when using RNA extracted samples obtained from nasopharyngeal swabs collected in guanidine-containing viral transport medium. Typical RT-LAMP reactions were performed at 65°C for 30 min. When compared to reverse transcriptase–quantitative polymerase chain reaction (RT-qPCR), up to cycle-threshold (Ct) value 32, RT-LAMP presented 98% [95% confidence interval (CI) = 95.3–99.5%] sensitivity and 100% (95% CI = 94.5–100%) specificity for SARS-CoV-2 RNA detection targeting E and N genes. No cross-reactivity was detected when testing other non–SARS-CoV virus, confirming high specificity. The test is compatible with primary RNA extraction–free samples. We also demonstrated that colorimetric RT-LAMP can detect SARS-CoV-2 variants of concern and variants of interest, such as variants occurring in Brazil named gamma (P.1), zeta (P.2), delta (B.1.617.2), B.1.1.374, and B.1.1.371. The method meets point-of-care requirements and can be deployed in the field for high-throughput COVID-19 testing campaigns, especially in countries where COVID-19 testing efforts are far from ideal to tackle the pandemics. Although RT-qPCR is considered the gold standard for SARS-CoV-2 RNA detection, it requires expensive equipment, infrastructure, and highly trained personnel. In contrast, RT-LAMP emerges as an affordable, inexpensive, and simple alternative for SARS-CoV-2 molecular detection that can be applied to massive COVID-19 testing campaigns and save lives.
Secretion of Gluconic Acid From Nguyenibacter sp. L1 Is Responsible for Solubilization of Aluminum Phosphate
Authors: Li Xiao Li , Zhao Xue Qiang , Dong Xiao Ying , Ma Jian Feng , Shen Ren Fang
Published: November 2021

Phosphorus (P) deficiency is one of the major factors limiting plant growth in acid soils, where most P is fixed by toxic aluminum (Al). Phosphate-solubilizing bacteria (PSBs) are important for the solubilization of fixed P in soils. Many PSBs have been isolated from neutral and calcareous soils, where calcium phosphate is the main P form, whereas PSBs in acid soils have received relatively little attention. In this study, we isolated a PSB strain from the rhizosphere of Lespedeza bicolor, a plant well adapted to acid soils. On the basis of its 16S rRNA gene sequence, this strain was identified as a Nguyenibacter species and named L1. After incubation of Nguyenibacter sp. L1 for 48 h in a culture medium containing AlPO4 as the sole P source, the concentration of available P increased from 10 to 225 mg L–1, and the pH decreased from 5.5 to 2.5. Nguyenibacter sp. L1 exhibited poor FePO4 solubilization ability. When the pH of non-PSB-inoculated medium was manually adjusted from 5.5 to 2.5, the concentration of available P only increased from 6 to 65 mg L–1, which indicates that growth medium acidification was not the main contributor to the solubilization of AlPO4 by Nguyenibacter sp. L1. In the presence of glucose, but not fructose, Nguyenibacter sp. L1 released large amounts of gluconic acid to solubilize AlPO4. Furthermore, external addition of gluconic acid enhanced AlPO4 solubilization and reduced Al toxicity to plants. We conclude that secretion of gluconic acid by Nguyenibacter sp. L1, which is dependent on glucose supply, is responsible for AlPO4 solubilization as well as the alleviation of Al phytotoxicity by this bacterial strain.
Molecular Mechanisms of the RECQ4 Pathogenic Mutations
Authors: Xu Xiaohua , Chang Chou-Wei , Li Min , Liu Chao , Liu Yilun
Published: November 2021

The human RECQ4 gene encodes an ATP-dependent DNA helicase that contains a conserved superfamily II helicase domain located at the center of the polypeptide. RECQ4 is one of the five RECQ homologs in human cells, and its helicase domain is flanked by the unique amino and carboxyl termini with sequences distinct from other members of the RECQ helicases. Since the identification of the RECQ4 gene in 1998, multiple RECQ4 mutations have been linked to the pathogenesis of three clinical diseases, which are Rothmund-Thomson syndrome, Baller-Gerold syndrome, and RAPADILINO. Patients with these diseases show various developmental abnormalities. In addition, a subset of RECQ4 mutations are associated with high cancer risks, especially for osteosarcoma and/or lymphoma at early ages. The discovery of clinically relevant RECQ4 mutations leads to intriguing questions: how is the RECQ4 helicase responsible for preventing multiple clinical syndromes? What are the mechanisms by which the RECQ4 disease mutations cause tissue abnormalities and drive cancer formation? Furthermore, RECQ4 is highly overexpressed in many cancer types, raising the question whether RECQ4 acts not only as a tumor suppressor but also an oncogene that can be a potential new therapeutic target. Defining the molecular dysfunctions of different RECQ4 disease mutations is imperative to improving our understanding of the complexity of RECQ4 clinical phenotypes and the dynamic roles of RECQ4 in cancer development and prevention. We will review recent progress in examining the molecular and biochemical properties of the different domains of the RECQ4 protein. We will shed light on how the dynamic roles of RECQ4 in human cells may contribute to the complexity of RECQ4 clinical phenotypes.
Cytokine and Complement Response in the Glaucomatous βB1-CTGF Mouse Model
Authors: Reinehr Sabrina , Doerner Johanna D. , Mueller-Buehl Ana M. , Koch Dennis , Fuchshofer Rudolf , Dick H. Burkhard , Joachim Stephanie C.
Published: November 2021

Glaucoma is a complex neurodegenerative disease leading to a loss of retinal ganglion cells (RGCs) and optic nerve axons. An activation of the complement system seems to contribute to cell loss in this disease. Hence, we investigated a possible initiation of the complement system and the cytokine response in the βB1-CTGF glaucoma model. In these mice, intraocular pressure is elevated, which is the main glaucoma risk factor in patients, and RGC loss occurs at 15 weeks of age. Therefore, quantitative real-time PCR and immunohistological experiments were performed in 5-, 10-, and 15-week-old βB1-CTGF animals and their corresponding wildtypes (WT) to analyze the expression of several complement system factors. We could show that mRNA levels of the terminal complement pathway components C3 and C5 (Hc) were upregulated at 10 weeks. In accordance, more C3+ and membrane attack complex+ cells were observed in transgenic retinae. Further, the C5a receptor anaphylatoxin receptor (C5ar) and the complement component C5a receptor 1 (C5ar1; CD88) mRNA levels were upregulated in 10- and 15-week-old βB1-CTGF mice. Interestingly, all three activation routes of the complement system were elevated in βB1-CTGF mice at some age. Especially C1q, as a marker of the classical pathway, was significantly increased at all investigated ages. Furthermore, mRNA expression levels of interferon-γ (Infg) were upregulated at 5 weeks, while Cxcl1 and Cxcl2 mRNA levels were upregulated at 10 and 15 weeks. The mRNA levels of the chemokines Cxcl10 were increased at all ages in βB1-CTGF mice. These results lead to the assumption that in these transgenic mice, a complement activation mainly through the classical pathway as well as a cytokine response plays a major role in cell death.
Asymptomatic Carotid Disease and Cognitive Impairment: What Is the Evidence?
Authors: Baradaran Hediyeh , Sarrami Amir Hossein , Gupta Ajay
Published: November 2021

The development of cognitive dysfunction and dementia is a complex, multifactorial process. One of the contributors to various types of cognitive dysfunction is carotid atherosclerosis which can frequently be seen in asymptomatic individuals. There are a number of different manifestations of asymptomatic carotid atherosclerosis including arterial stiffness, carotid intima-media thickening, flow-limiting stenosis, and complex, atherosclerotic plaque. Each of these forms of atherosclerosis may contribute to cerebral parenchymal damage, contributing to cognitive dysfunction. In this review article, we will discuss each of these forms of carotid atherosclerosis, present the potential mechanistic underpinnings behind an association, and then review the scientific evidence supporting potential associations to cognitive dysfunction and dementia.