bluuebunny/crossref_metadata_2025 huggingface dataset:
Viral metagenomics revealed diverse CRESS-DNA virus genomes in faeces of forest musk deer
Authors: Liu Qi , Wang Hao , Ling Yu , Yang Shi-Xing , Wang Xiao-Chun , Zhou Rui , Xiao Yu-Qing , Chen Xu , Yang Jie , Fu Wei-Guo , Zhang Wen , Qi Gui-Lan
Published: April 2020

Background

Musk deer can produce musk which has high medicinal value and is closely related to human health. Viruses in forest musk deer both threaten the health of forest musk deer and human beings.

Methods

Using viral metagenomics we investigated the virome in 85 faeces samples collected from forest musk deer.

Results

In this article, eight novel CRESS-DNA viruses were characterized, whole genomes were 2148 nt–3852 nt in length. Phylogenetic analysis indicated that some viral genomes were part of four different groups of CRESS-DNA virus belonging in the unclassified CRESS-DNA virus, Smacoviridae, pCPa-like virus and pPAPh2-like virus. UJSL001 (MN621482), UJSL003 (MN621469) and UJSL017 (MN621476) fall into the branch of unclassified CRESS-DNA virus (CRESSV1–2), UJSL002 (MN621468), UJSL004 (MN621481) and UJSL007 (MN621470) belong to the cluster of Smacoviridae, UJSL005 (MN604398) showing close relationship with pCPa-like (pCRESS4–8) clusters and UJSL006 (MN621480) clustered into the branch of pPAPh2-like (pCRESS9) virus, respectively.

Conclusion

The virome in faeces samples of forest musk deer from Chengdu, Sichuan province, China was revealed, which further characterized the diversity of viruses in forest musk deer intestinal tract.
“The plants have axé”: investigating the use of plants in Afro-Brazilian religions of Santa Catarina Island
Authors: Pagnocca Tiago Santos , Zank Sofia , Hanazaki Natalia
Published: April 2020

Background

Cultural and religious practices of African origin have decisively influenced traditional health practices in the Americas since the African diaspora. Plants are core elements in the religions of African origin. Compared with other parts of Brazil where the Afro-Brazilian presence is widely recognized, in Southern Brazil, these cultural practices are often socially invisible. Yet, there are several terreiros of three Afro-Brazilian religions: Candomblé, Umbanda, and Ritual deAlmas e Angola. We hypothesize that the importance of plants in Afro-Brazilian religions is linked not only to spiritual and magical issues but also to the medicinal properties of these plants. We seek to answer the following questions: (a) Which plants are used in the terreiros and what are their indications for use?; (b) Are there plants that stand out culturally in these religious groups?; and (c) What is the importance of the adaptive maintenance and replacement process in the use of plants in these religions, considering the Neotropical and African plants?

Methods

We performed a census of the existing terreiros on the Island of Santa Catarina to collect information on the knowledge and use of plants. In all terreiros that consented to participate in the research, we collected data through semi-structured interviews, guided tours for plant collection, and participant observation. We identified the botanical species through expert consultations and botanical literature.

Results

We interviewed 27 spiritual leaders, who cited 93 plants belonging to 86 botanical species. We identified 14 categories of use, with emphasis on liturgical ritual use (59%), general and unspecified diseases (32%), and digestive diseases (27%). In most liturgics uses, direct contact between plant and patient occurs, as in the case of bathing and the cleansing use of smoke. Sixteen plants were cited in all terreiros, configuring a set of species that can be considered as culturally important plants for these religious groups.

Conclusions

These groups have extensive knowledge about a highly consensual set of therapeutic plants that should be further investigated pharmacologically to understand the effect of their external use. Also, we emphasize the importance of recognizing and valuing this ancestral Afro-Brazilian knowledge and learning also from these people about their broader vision of health which also adds more spirituality in health care.
Fauna and conservation in the context of formal education: a study of urban and rural students in the semi-arid region of Brazil
Authors: de Oliveira José Valberto , da Silva Moacyr Xavier Gomes , Borges Anna Karolina Martins , Souto Wedson Medeiros Silva , de Faria Lopes Sérgio , de Brito Melo Trovão Dilma Maria , Barboza Raynner Rilke Duarte , Alves Rômulo Romeu Nóbrega
Published: April 2020

Background

In addition to playing a key role in the dynamics of ecosystems, animal diversity, especially that of wild vertebrates, is intimately linked with human evolutionary history, which has resulted in diverse interactions that must be emphasized in formal education processes. We analyzed several methods of approaches used for biological education in order to teach about wild vertebrates and their conservation in urban and rural schools in the semi-arid region of Brazil.

Methods

Data were obtained via questionnaires applied to 990 students, of which 528 were urban and 462 rural, distributed among the seven grades/years that comprise the last two cycles of basic education in Brazil. The richness and diversity of the animals cited by the students were calculated, being the diversity using an adaptation of the equation for the Shannon-Weaver Index (H′). Data were analyzed using non-parametric descriptive statistics.

Results

Mammals and birds had the greatest richness and diversity of animals cited as most-studied in science/biology classes, and also the most indicated as occurring in the studied region. Among mammals, large carnivores with a showy appearance and utilitarian value had the highest citation frequencies, while there was a tendency for limited recognition of faunistic diversity in the other groups mentioned. Almost 70% of the students stated that their schooling processes dealt with the conservation of wild animals; however, about 50% of the students in both urban and rural contexts did not express conceptual understanding about the conservation of nature.

Conclusions

The recognition of animal diversity, especially vertebrates, beyond just mammals and birds, as well as conceptual clarity about the conservation of nature, are fundamental factors for the development of critical awareness of fauna and its conservation, and where the processes of schooling have a preponderant role. Finally, the study contributes to the legitimization of Ethnobiology as an interdisciplinary field of knowledge, especially in its interface with education, in addition to pointing out the importance of optimizing efforts in approaches to biodiversity conservation in formal educational processes.
Identification of stable reference genes for qPCR studies in common wheat (Triticum aestivum L.) seedlings under short-term drought stress
Authors: Dudziak Karolina , Sozoniuk Magdalena , Szczerba Hubert , Kuzdraliński Adam , Kowalczyk Krzysztof , Börner Andreas , Nowak Michał
Published: April 2020

Background

Quantitative PCR (qPCR) is one of the most common and accurate methods of gene expression analysis. However, the biggest challenge for this kind of examinations is normalization of the results, which requires the application of dependable internal controls. The selection of appropriate reference genes (RGs) is one of the most crucial points in qPCR data analysis and for correct assessment of gene expression. Because of the fact that many reports indicate that the expression profiles of typically used RGs can be unstable in certain experimental conditions, species or tissues, reference genes with stable expression levels should be selected individually for each experiment. In this study, we analysed a set of ten candidate RGs for wheat seedlings under short-term drought stress. Our tests included five ‘traditional’ RGs (GAPDH, ACT, UBI, TUB, and TEF1) and five novel genes developed by the RefGenes tool from the Genevestigator database.

Results

Expression stability was assessed using five different algorithms: geNorm, NormFinder, BestKeeper, RefFinder and the delta Ct method. In the final ranking, we identified three genes: CJ705892, ACT, and UBI, as the best candidates for housekeeping genes. However, our data indicated a slight variation between the different algorithms that were used. We revealed that the novel gene CJ705892, obtained by means of in silico analysis, showed the most stable expression in the experimental tissue and condition.

Conclusions

Our results support the statement, that novel genes selected for certain experimental conditions have a more stable level of expression in comparison to routinely applied RGs, like genes encoding actin, tubulin or GAPDH. Selected CJ705892 gene can be used as a housekeeping gene in the expression analysis in wheat seedlings under short-term drought. The results of our study will be useful for subsequent analyses of gene expression in wheat tissues subjected to drought.
The combinations of multiple factors to improve the diagnostic sensitivity and specificity after artificial joint infection
Authors: Chu Lei , Ren You-Liang , Yang Jun-Song , Yang Jin , Zhou Hang , Jiang Hai-Tao , Shi Lei , Hao Ding-Jun , Deng Zhong-Liang
Published: April 2020

Objective

To discuss the sensitivity and specificity of the combinations of multiple factors that work on bone infection after artificial joint, and provide evidence-based medical basis for the early diagnosis of infection after artificial joint.

Methods

A retrospective review was conducted on 35 patients diagnosed with periprosthetic joint infections (PJI) or aseptic loosening (AL) who both received revision operation from January 2011 to January 2015. Analyzing and comparing their epidemiology indexes and expounded a series of auxiliary examinations corresponding positive diagnosis ratio.

Results

Thirty-five patients were divided into two groups. One is called group PJI which includes 16 patients, and the other is called group AL which contains 19 patients. There was no statistical difference between in age (p= 0.536), gender ratio (p= 0.094), and the time of catching infection or getting loose (p= 0.055). Swelling was statistical significant (p= 0.0435 < 0.05). AUC of CRP = 0.947, ESR = 0.893, IL-6 = 0.893, PCT = 0.781, WBC = 0.839, and PMN = 0.755, respectively, CRP has a high diagnostic value to PJI, ESR, IL-6, PCT, WBC, and PMN% possess a moderate diagnostic value. There were 3 cases of PJI whose pathological paraffin section showed infectious inflammatory cells (100%). three PJI patients and one AL patient whose 99mTc-MDP examination presented 100% infection or looseness rate.

Conclusion

CRP has a high diagnostic value to PJI. Histopathology HE staining, Gram staining, and 99mTc-MDP provide a highly accurate diagnosis for PJI. Therefore, the results suggest combining the unique clinical symptoms of PJI patients with relevant laboratory indexes, histopathologic characteristics, and imageological examinations that can improve diagnostic sensitivity and specificity of PJI in its early stage.
PENK inhibits osteosarcoma cell migration by activating the PI3K/Akt signaling pathway
Authors: Zhang Hai-ping , Yu Zi-liang , Wu Bing-bing , Sun Fa-rui
Published: April 2020

Background

This article reports the effects of proenkephalin (PENK) on osteosarcoma (OS) cell migration.

Methods

A Gene Expression Omnibus (GEO) dataset was used to identify differentially expressed genes (DEGs) in OS tumor samples and normal human osteoblasts. Tumor tissue and adjacent normal tissue were collected from 40 OS patients. MG63 cells were transfected with si-PENK. Transwell migration assays and wound healing assays were performed to compare the effect of PENK on migration. Moreover, LY294002 was used to identify the potential mechanism. Gene expression was examined via qRT-PCR and Western blotting.

Results

Bioinformatic analysis revealed that PENK was downregulated in OS tumor samples compared with normal human osteoblasts. Moreover, PENK was identified as the hub gene of the DEGs. The PI3K/Akt signaling pathway was significantly enriched in the DEGs. Moreover, PENK was downregulated in OS and MG63 cells compared with the corresponding control cells. Silencing PENK promoted MG63 cell migration; however, treatment with LY294002 partially attenuated PENK silencing-induced OS cell migration.

Conclusion

PENK inhibits OS cell migration by activating the PI3K/Akt signaling pathway.
Effects of iron chelation therapy on the clinical course of aceruloplasminemia: an analysis of aggregated case reports
Authors: Vroegindeweij Lena H. P. , Boon Agnita J. W. , Wilson J. H. Paul , Langendonk Janneke G.
Published: April 2020

Background

Aceruloplasminemia is a rare genetic iron overload disorder, characterized by progressive neurological manifestations. The effects of iron chelation on neurological outcomes have only been described in case studies, and are inconsistent. Aggregated case reports were analyzed to help delineate the disease-modifying potential of treatment.

Methods

Data on clinical manifestations, treatment and neurological outcomes of treatment were collected from three neurologically symptomatic Dutch patients, who received deferiprone with phlebotomy as a new therapeutic approach, and combined with other published cases. Neurological outcomes of treatment were compared between patients starting treatment when neurologically symptomatic and patients without neurological manifestations.

Results

Therapeutic approaches for aceruloplasminemia have been described in 48 patients worldwide, including our three patients. Initiation of treatment in a presymptomatic stage of the disease delayed the estimated onset of neurological manifestations by 10 years (median age 61 years, SE 5.0 vs. median age 51 years, SE 0.6, p = 0.001). Although in 11/20 neurologically symptomatic patients neurological manifestations remained stable or improved during treatment, these patients were treated significantly shorter than patients who deteriorated neurologically (median 6 months vs. median 43 months, p = 0.016). Combined iron chelation therapy with deferiprone and phlebotomy for up to 34 months could be safely performed in our patients without symptomatic anemia (2/3), but did not prevent further neurological deterioration.

Conclusions

Early initiation of iron chelation therapy seems to postpone the onset of neurological manifestations in aceruloplasminemia. Publication bias and significant differences in duration of treatment should be considered when interpreting reported treatment outcomes in neurologically symptomatic patients. Based on theoretical grounds and the observed long-term safety and tolerability in our study, we recommend iron chelation therapy with deferiprone in combination with phlebotomy for aceruloplasminemia patients without symptomatic anemia.
Telemedicine strategy of the European Reference Network ITHACA for the diagnosis and management of patients with rare developmental disorders
Authors:   on behalf of ERN ITHACA, Smith Michael , Alexander Elizabeth , Marcinkute Ruta , Dan Dorica , Rawson Myfanwy , Banka Siddharth , Gavin Jason , Mina Hany , Hennessy Con , Riccardi Florence , Radio Francesca Clementina , Havlovicova Marketa , Cassina Matteo , Emandi Adela Chirita , Fradin Melanie , Gompertz Lianne , Nordgren Ann , Traberg Rasa , Rossi Massimiliano , Trimouille Aurelién , Sowmyalakshmi Rasika , Dallapiccola Bruno , Renieri Alessandra , Faivre Laurence , Kerr Bronwyn , Verloes Alain , Clayton-Smith Jill , Douzgou Sofia
Published: April 2020

Background

The European Reference Networks, ERNs, are virtual networks for healthcare providers across Europe to collaborate and share expertise on complex or rare diseases and conditions. As part of the ERNs, the Clinical Patient Management System, CPMS, a secure digital platform, was developed to allow and facilitate web-based, clinical consultations between submitting clinicians and relevant international experts. The European Reference Network on Intellectual Disability, TeleHealth and Congenital Anomalies, ERN ITHACA, was formed to harness the clinical and diagnostic expertise in the sector of rare, multiple anomaly and/or intellectual disability syndromes, chromosome disorders and undiagnosed syndromic disorders. We present the first year results of CPMS use by ERN ITHACA as an example of a telemedicine strategy for the diagnosis and management of patients with rare developmental disorders.

Results

ERN ITHACA ranked third in telemedicine activity amongst 24 European networks after 12 months of using the CPMS. Information about 28 very rare cases from 13 different centres across 7 countries was shared on the platform, with diagnostic or other management queries. Early interaction with patient support groups identified data protection as of primary importance in adopting digital platforms for patient diagnosis and care. The first launch of the CPMS was built to accommodate the needs of all ERNs. The ERN ITHACA telemedicine process highlighted a need to customise the CPMS with network-specific requirements. The results of this effort should enhance the CPMS utility for telemedicine services and ERN-specific care outcomes.

Conclusions

We present the results of a long and fruitful process of interaction between the ERN ITHACA network lead team and EU officials, software developers and members of 38 EU clinical genetics centres to organise and coordinate direct e-healthcare through a secure, digital platform. The variability of the queries in just the first 28 cases submitted to the ERN ITHACA CPMS is a fair representation of the complexity and rarity of the patients referred, but also proof of the sophisticated and variable service that could be provided through a structured telemedicine approach for patients and families with rare developmental disorders. Web-based approaches are likely to result in increased accessibility to clinical genomic services.
Low occurrence of extended-spectrum cephalosporinase producing Enterobacteriaceae and no detection of methicillin-resistant coagulase-positive staphylococci in healthy dogs in Sweden
Authors: Börjesson Stefan , Gunnarsson Lotta , Landén Annica , Grönlund Ulrika
Published: April 2020

Sweden has a long tradition of monitoring occurrence of antibiotic resistant bacteria in both animals and humans, but there currently is no organised and harmonized monitoring on carriage of Enterobacteriaceae producing extended-spectrum beta-lactamase (ESBL), plasmid-mediated AmpC beta-lactamase (pAmpC), or methicillin-resistant coagulase positive staphylococci e.g. methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus pseudintermedius (MRSP) in dogs. The aim of the current study was therefore to determine the prevalence of ESBL/pAmpC producing Enterobacteriaceae and methicillin-resistant coagulase positive staphylococci in healthy dogs in Sweden, and to phenotypically and genotypically characterize any identified isolates. It was shown that 0.9% (95% confident interval 0.3–2.7%) of the dogs (n = 325) carried multi-resistant ESBL-producing Escherichia coli, but that no methicillin-resistant coagulase positive staphylococci could be detected. In conclusion, the occurrence of multi-drug resistant bacteria remains rare among healthy dogs in Sweden. In addition, the ESBL-producing E. coli identified showed genetic characteristics related to those reported from humans.
Acute mechanical stress in primary porcine RPE cells induces angiogenic factor expression and in vitro angiogenesis
Authors: Farjood Farhad , Ahmadpour Amir , Ostvar Sassan , Vargis Elizabeth
Published: April 2020

Background

Choroidal neovascularization (CNV) is a major cause of blindness in patients with age-related macular degeneration. CNV is characterized by new blood vessel growth and subretinal fluid accumulation, which results in mechanical pressure on retinal pigment epithelial (RPE) cells. The overexpression of RPE-derived angiogenic factors plays an important role in inducing CNV. In this work, we investigated the effect of mechanical stress on the expression of angiogenic factors in porcine RPE cells and determined the impact of conditioned medium on in-vitro angiogenesis.

Results

The goal of this study was to determine whether low levels of acute mechanical stress during early CNV can induce the expression of angiogenic factors in RPE cells and accelerate angiogenesis. Using a novel device, acute mechanical stress was applied to primary porcine RPE cells and the resulting changes in the expression of major angiogenic factors, VEGF, ANG2, HIF-1α, IL6, IL8 and TNF-α, were examined using immunocytochemistry, qRT-PCR, and ELISA. An in vitro tube formation assay was used to determine the effect of secreted angiogenic proteins due to mechanical stress on endothelial tube formation by human umbilical vein endothelial cells (HUVECs). Our results showed an increase in the expression of VEGF, ANG2, IL-6 and IL-8 in response to mechanical stress, resulting in increased in vitro angiogenesis. Abnormal epithelial-mesenchymal transition (EMT) in RPE cells is also associated with CNV and further retinal degeneration. Our qRT-PCR results verified an increase in the expression of EMT genes, CDH2, VIM and FN1, in RPE cells.

Conclusions

In conclusion, we showed that acute mechanical stress induces the expression of major angiogenic and EMT factors and promotes in vitro angiogenesis, suggesting that mechanical stress plays a role in promoting aberrant angiogenesis in AMD.
A network pharmacology-based study on Alzheimer disease prevention and treatment of Qiong Yu Gao
Authors: You Jie-shu , Li Chen-yue , Chen Wei , Wu Xia-lin , Huang Li-jie , Li Ren-kai , Gao Fei , Zhang Ming-yue , Liu Huan-lan , Qu Wei-ling
Published: April 2020

Background and objective

As the pathological mechanisms of AD are complex, increasing evidence have demonstrated Chinese Medicine with multi-ingredients and multi-targets may be more suitable for the treatment of diseases with complex pathogenesis. Therefore, the study was to preliminarily decipher the bioactive compounds and potential mechanisms of Qiong Yu Gao (QYG) for AD prevention and treatment by an integrated network pharmacology approach.

Methods

Putative ingredients of QYG and significant genes of AD were retrieved from public database after screening. Then QYG ingredients target proteins/genes were obtained by target fishing. Compound-target-disease network was constructed using Cytoscape to decipher the mechanism of QYG for AD. KEGG pathway and GO enrichment analysis were performed to investigate the molecular mechanisms and pathways related to QYG for AD treatments.

Results

Finally, 70 compounds and 511 relative drug targets were collected. In which, 17 representative direct targets were found. Gene ontology enrichment analysis revealed that the adenylate cyclase-inhibiting G-protein coupled acetylcholine receptor signaling pathway was the key biological processes and were regulated simultaneously by the 17 direct targets. The KEGG pathway enrichment analysis found that three signaling pathways were closely related to AD prevention and treatment by QYG, including PI3K-Akt signaling pathway, regulation of actin cytoskeleton pathway and insulin resistance pathway.

Conclusion

This study demonstrated that QYG exerted the effect of preventing and treating AD by regulating multi-targets with multi-components. Furthermore, the study demonstrated that a network pharmacology-based approach was useful for elucidation of the interrelationship between complex diseases and interventions of Chinese herbal medicines.
Long non-coding RNA AOC4P suppresses epithelial ovarian cancer metastasis by regulating epithelial-mesenchymal transition
Authors: Lin Xiaojing , Tang Xiaoyan , Zheng Tingting , Qiu Junjun , Hua Keqin
Published: April 2020

Objective

Currently, the function and mechanisms of long non-coding RNAs (lncRNAs) involved in the metastasis of epithelial ovarian cancer (EOC), especially those of the lncRNAs participated in the epithelial-mesenchymal transition (EMT) process, remains largely unknown. Here, we focused on a lncRNA named AOC4P and analysed its role in EOC.

Materials and methods

The expression of AOC4P gene was examined with quantitative real-time quantitative PCR (qRT-PCR). The cell migration and invasion were detected by Transwell and scratch assays. The in vivo metastatic activity was evaluated by intraperitoneal metastasis model. The downstream genes were investigated by a tumour EMT real-time polymerase chain reaction (RT-PCR) array, and validated by qRT-PCR and Western blot.

Results

The results showed that AOC4P expression levels were decreased in EOC tissues and cell lines, and that the under-expression of AOC4P was positively correlated with FIGO stage and lymph node metastasis. Furthermore, the knockdown of AOC4P expression in poorly metastatic EOC cell lines remarkably facilitated cell migration/invasion while the overexpression of AOC4P in highly metastatic EOC cell lines reduced the metastatic ability of these cells in vitro. Consistently, the anti-metastatic role of AOC4P in vivo was also verified by bioluminescence imaging and tumour dissection. Mechanistically, the anti-metastatic effect of AOC4P in EOC was partially mediated by the EMT process accompanied by the alterations in MMP9 and COL1A2 expression.

Conclusion

These data highlight that AOC4P plays a critical role in EOC invasion/metastasis and could function as a novel and effective target for the lncRNA-based anti-metastatic clinical management of EOC.
A study on the antioxidant and antimicrobial activities in the chloroformic and methanolic extracts of 6 important medicinal plants collected from North of Iran
Authors: Hadadi Zahra , Nematzadeh Ghorban Ali , Ghahari Somayeh
Published: April 2020

Background

As possible sources of natural bioactive molecules, the plant essential oils and extracts have been used globally in new antimicrobial compounds, food preservatives, and alternatives to treat infectious disease.

Methods

In this research, the antimicrobial activities of chloroformic and methanolic extracts of Sophora flavescens, Rhaponticum repens, Alhagi maurorum, Melia azedarach, Peganum harmala, and Juncus conglomeratus were evaluated against 8 bacteria (S. aureus, B. subtilis, R. toxicus, P. aeruginosa, E. coli, P. syringae, X. campestris, P. viridiflava) and 3 fungi (Pyricularia oryzae, Fusarium oxysporum and Botrytis cinerea), through disc diffusion method. Furthermore, the essential oils of plants with the highest antibacterial activity were analyzed utilizing GC/MS. Moreover, the tested plants were exposed to screening for possible antioxidant effect utilizing DPPH test, guaiacol peroxidas, and catalase enzymes. Besides, the amount of total phenol and flavonoid of these plants was measured.

Results

Among the tested plants, methanolic and chloroformic extracts of P. harmala fruits showed the highest antibacterial activity against the tested bacteria. Besides, the investigation of free radical scavenging effects of the tested plants indicated the highest DPPH, protein, guaiacol peroxidase, and catalase in P. harmala, M. azedarach, J. conglomeratus fruits, and J. conglomeratus fruits, respectively. In addition, the phytochemical analysis demonstrated the greatest amounts of total phenolic and flavonoid compositions in J. conglomeratus and P. harmala, respectively.

Conclusion

The results indicated that these plants could act as a promising antimicrobial agent, due to their short killing time.
Primary validation of Charm II tests for the detection of antimicrobial residues in a range of aquaculture fish
Authors: Mukota Aziz Kimera , Gondam Melanie Flore Kamini , Tsafack Julie Judith Takadong , Sasanya James , Reybroeck Wim , Ntale Muhammad , Nyanzi Steven Allan , Tebandeke Emmanuel
Published: April 2020

The study carried out a primary validation of Charm II tests for the detection of antimicrobial residues in aquaculture fish. The validation was performed according to European Commission Decision 2002/657/EC and the parameters determined included: detection capability, repeatability, reproducibility, specificity and robustness for the detection of antimicrobial residues in fish. Fish materials from different species including cat fish, trout, salmon, sea bass, tilapia, lingue and pangasius, were spiked with varying concentrations of selected antimicrobials including sulfonamides, β-lactams, macrolides, tetracyclines and aminoglycosides to determine the detection capabilities and other validation parameters of the Charm II tests. Results of the validation showed that the detection capabilities for the tetracyclines ranged from 25 to 100 µg/kg, while the sulfonamides and aminoglycosides were detected at 25 µg/kg for all species under study. The detection capabilities for the beta-lactams ranged from 25 to 300 µg/kg; and was 100 µg/kg for the tested macrolides. Results of the study showed that there was no significant difference between counts for samples read immediately after addition of the scintillation liquid and those read 14 h after addition of the scintillation liquid, provided that there was good vortexing before analysis. There was also no significant difference between counts for the same samples analyzed in different runs under repeatability and reproducibility conditions at the same spiking concentrations for the different fish species analyzed. The relative standard deviation for both repeatability and reproducibility ranged from 1.2 to 15.1%. The Charm II tests were found to be 100% group specific, as none of the antimicrobials kits, gave false positive results when testing non-target antimicrobial drugs. Results of this study demonstrate the suitability of the Charm II technique as a rapid screening tool for detection of antimicrobial residues in a variety of fish species at maximum residue limits (MRL) established in the EU guidelines, with the exception of tilmicosin which was detected at 2 MRL. The results also prove the robustness, specificity, reliability and precision of the Charm II assay in the detection of various antimicrobial residuals in fish and its applicability for the rapid evaluation of the quality of aquaculture fish for safety and trade purposes.
Vacuolar ATPase depletion contributes to dysregulation of endocytosis in bloodstream forms of Trypanosoma brucei
Authors: Xu Zhi-Shen , Li Feng-Jun , Hide Geoff , Lun Zhao-Rong , Lai De-Hua
Published: April 2020

Background

Vacuolar H+-ATPase (V-ATPase) is a highly conserved protein complex which hydrolyzes ATP and pumps protons to acidify vacuolar vesicles. Beyond its role in pH maintenance, the involvement of V-ATPase in endocytosis is well documented in mammals and plants but is less clear inTrypanosoma brucei.

Methods

In this study, the subcellular localization of V-ATPase subunit B (TbVAB) ofT. bruceiwas assessedvia in situN-terminal YFP-tagging and immunofluorescence assays. Transgenic bloodstream forms (BSF) ofT. bruceiwere generated which comprised either a V-ATPase subunit B (TbVAB) conditional knockout or a V-ATPase subunit A (TbVAA) knockdown. Acridine orange and BCECF-AM were employed to assess the roles of V-ATPase in the pH regulation of BSFT. brucei. The endocytic activities of three markers were also characterized by flow cytometry analyses. Furthermore, trypanosomes were counted from trypanolysis treatment groups (either containing 1% or 5% NHS) and endocytosed trypanosome lytic factor (TLF) was also analyzed by an immunoblotting assay.

Results

TbVAB was found to localize to acidocalcisomes, lysosomes and probably also to endosomes of BSF ofT. bruceiand was demonstrated to be essential for cell growth.TbVABdepletion neutralized acidic organelles at 24 hours post-tetracycline depletion (hpd), meanwhile the steady state intracellular pH increased from 7.016 ± 0.013 to 7.422 ± 0.058. Trypanosomes withTbVABdepletion at 24 hpd were found to take up more transferrin (2.068 ± 0.277 fold) but less tomato lectin (49.31 ± 22.57%) by endocytosis, while no significant change was detected in dextran uptake. Similar endocytic dysregulated phenotypes were also observed inTbVAAknockdown cells. In addition,TbVABdepleted trypanosomes showed a low uptake of TLF and exhibited less sensitive to lysis in both 1% and 5% NHS treatments.

Conclusions

TbVAB is a key component of V-ATPase and was found to play a key function in endocytosis as well as exhibiting different effects in a receptor/cargo dependent manner in BSF ofT. brucei. Besides vacuolar alkalinization, the dysregulation of endocytosis inTbVABdepletedT. bruceiis considered to contribute to the reduced sensitivity to lysis by normal human serum.
Clonorchis sinensis secretory protein CsAg17 vaccine induces immune protection
Authors: Bai Xuelian , Song Jin-Ho , Dai Fuhong , Lee Ji-Yun , Hong Sung-Jong
Published: April 2020

Background

Clonorchiasis is endemic in East and Southeast Asian countries. For a preventive strategy against infectious diseases, vaccination is the most effective. Here, we evaluated the molecular characteristics and immune responses of CsAg17 protein fromClonorchis sinensis, and investigated its protective effects againstC. sinensischallenge.

Methods

A cDNA clone encoding CsAg17 protein and containing a secretory signal peptide at the N-terminus was retrieved from theC. sinensistranscriptome bank. Recombinant CsAg17 B-cell epitope protein and cDNA vaccines were produced and their immune responses were evaluated in FVB mice. The proportional changes of CD3+/CD4+and CD3+/CD8+T cells were detected by flow cytometry, and immune effectors were measured by ELISA.

Results

TheCsAg17mRNA was transcribed at a higher level inC. sinensisadults than in metacercariae. The CsAg17 protein was distributed in the sperms, oral and ventral suckers, and mesenchymal tissues ofC. sinensisadults. In mice challenged withC. sinensismetacercariae, vaccination with CsAg17 protein and cDNA resulted in a reduction to 64% and 69% in worm burden, respectively. Both CsAg17 protein and cDNA vaccines increased the proportion of CD3+/CD4+and CD3+/CD8+T cells and stimulated the production of Th1 type cytokines such as interleukin (IL)-2, IL-12, and interferon-γ, while maintaining minimum levels of Th2 cytokines. The levels of IgG specific to CsAg17 protein steeply increased in the two vaccinated groups from 2 weeks after immunization. The liver tissue retained good morphology in the mice vaccinated with CsAg17 protein or cDNA, whereas severe inflammation and large serous cysts were observed in the liver of the unvaccinated mice.

Conclusions

Vaccination with CsAg17 protein and cDNA reduced the pathological changes in the bile duct and liver, and ameliorated the worm burdenviacellular and humoral immune responses. Thus, they may serve as good vaccine candidates againstC. sinensisinfections.
Detection of Cercopithifilaria bainae infection in shelter dogs and ticks in Oklahoma, USA
Authors: Lineberry Megan W. , Sundstrom Kellee D. , Little Susan E. , Stayton Erin M. , Allen Kelly E.
Published: April 2020

Background

Cercopithifilaria bainae is a filarioid nematode of dogs. Infection with the parasite was not reported in the USA until 2017, when a dog with skin lesions in Florida was diagnosed. Brown dog ticks, Rhipicephalus sanguineus (sensu lato), are the purported tick vectors, and are widespread in the USA. Therefore, C. bainae is likely present in additional states. Here, we tested dogs and ticks in Oklahoma for evidence of C. bainae infection.

Methods

Dermal punch biopsies were opportunistically collected from municipal shelter and client-owned dogs. Multiple skin samples collected from interscapular and head regions were tested by saline sedimentation to recover live microfilariae for morphometric identification and by PCR to amplify a 330 bp region of the filarioid 12S rRNA gene. Also, ticks observed on surveyed dogs were collected, identified to species level, and tested for filarioid DNA.

Results

A total of 496 saline sedimentations were performed on 230 shelter and 20 client-owned dogs. Cercopithifilaria bainae infections were identified in 2.6% (6/230) of shelter dogs by morphometry of microfilariae in sedimentations and/or amplification of DNA from skin. DNA sequences amplified from PCR positive skin samples were 99–100% identical to C. bainae reported in Italy. All skin samples from client-owned dogs were negative for filarioid infection by saline sedimentation and PCR. A total of 112 ticks, comprised of four species, were collected. Two of 72 R. sanguineus (s.l.), both engorged females found attached to a C. bainae infected dog, harbored C. bainae DNA (99–100% identity). One attached R. sanguineus (s.l.) male on the same dog harbored filarioid DNA sequence which was difficult to interpret at numerous base-pair locations, but was closest in identity (~80%) to C. bainae.

Conclusions

The distribution of C. bainae is more widespread than previously known. To our knowledge, we document C. bainae infections in dogs and DNA in brown dog ticks in Oklahoma for the first time. As brown dog ticks are commonly found throughout the USA, veterinarians in this region should consider C. bainae infection as a differential diagnosis in canine patients with dermatitis or polyarthritis.
Higher interferon score and normal complement levels may identify a distinct clinical subset in children with systemic lupus erythematosus
Authors: Tesser Alessandra , de Carvalho Luciana Martins , Sandrin-Garcia Paula , Pin Alessia , Pastore Serena , Taddio Andrea , Roberti Luciana Rodrigues , de Paula Queiroz Rosane Gomes , Ferriani Virginia Paes Leme , Crovella Sergio , Tommasini Alberto
Published: April 2020

Background

Systemic lupus erythematosus (SLE) is a complex multi-system disease, characterized by both autoimmune and autoinflammatory clinical and laboratory features. The role of type I interferon (IFN) in SLE has been demonstrated from the 2000s, by gene expression analyses showing significant over-expression of genes related to type I IFN signalling pathway (IFN signature). However, several studies questioned the role of measuring the intensity of IFN signature (IFN score) to chase SLE activity.

We would assess if the IFN signature can help the clinical and therapeutic stratification of patients with pediatric SLE.

Methods

We measured the IFN score in peripheral whole blood from a series of subjects with childhood-onset SLE and correlated the results with clinical and laboratory parameters.

Results

Thirty-one subjects were included in the study, among which the 87% displayed a positive IFN score. The only significant relation was found for high IFN score in subjects with normocomplementemia. No correlation was observed between IFN score and SLEDAI-2K, BILAG-2004 and SLICC. Patients with high IFN score and normal complement levels also presented lower anti-dsDNA antibodies.

Conclusions

The integration between IFN signature analysis and complement levels may easily distinguish two groups of subjects, in which the autoimmune or autoinflammatory component of the disease seems to be prevalent.
Synergistic activation of NF-κB by TNFAIP3 (A20) reduction and UBE2L3 (UBCH7) augment that synergistically elevate lupus risk
Authors: Kim Taehyeung , Bae Sang-Cheol , Kang Changwon
Published: April 2020

Background

Systemic lupus erythematosus (SLE) is an autoimmune inflammatory rheumatic disease. SLE susceptibility is affected by multiple genetic elements, environmental factors, and their interactions. We aimed in this study to statistically and functionally characterize a gene-gene interaction (epistasis) recently documented to affect SLE risk.

Methods

Two single-nucleotide polymorphisms, rs2230926 in TNFAIP3 (A20) gene and rs131654 in UBE2L3 (UBCH7) gene, were genotyped in all 3525 Korean participants, and their SLE risk association and epistasis were statistically analyzed by calculating odds ratio (OR), 95% confidence interval (CI), and P values in genotype comparisons between 1318 SLE patients and 2207 healthy controls. Furthermore, their effects on gene functions were assessed by comparatively examining separate and combined effects of TNFAIP3 and UBE2L3 knockdowns on NF-κB transcription factor activity in human cells.

Results

SLE susceptibility is associated with TNFAIP3 rs2230926 (OR = 1.9, 95% CI 1.6–2.4, P = 8.6 × 10−11) and UBE2L3 rs131654 (OR = 1.2, 95% CI 1.1–1.4, P = 1.1 × 10−4) in a Korean population of this study. Their risk-associated alleles synergistically elevate SLE susceptibility in both multivariate logistic regression analysis (ORinteraction = 1.6, P = 0.0028) and genotype-stratified analysis (ORinteraction = 2.4), confirming the synergistic TNFAIP3-UBE2L3 interaction in SLE risk. Additionally, the SLE-susceptible alleles confer decreased TNFAIP3 expression (P = 1.1 × 10−6, n = 610) and increased UBE2L3 expression (P = 9.5 × 10−11, n = 475), respectively, in B cell analysis of the International HapMap Project individuals with adjustment for ethnicity. Furthermore, when compared with TNFAIP3 non-knockdown and UBE2L3 knockdown in human HeLa cells, TNFAIP3 knockdown and UBE2L3 non-knockdown synergistically increase three cytokines, CCL2, CXCL8 (IL8), and IL6, all regulated by NF-κB in the human TNFR signaling pathway.

Conclusions

A synergistic interaction between TNFAIP3 and UBE2L3 genes is observed in SLE risk, as being evident in comparison of genotype distributions between SLE patients and controls. Additionally, the synergistic gene-gene interaction is functionally validated, as TNFAIP3 reduction and UBE2L3 augment exert synergism in activation of NF-κB and subsequent induction of inflammatory cytokines. Accordingly, SLE inflammation and risk could be synergistically alleviated by TNFAIP3 upregulation and UBE2L3 downregulation.
Local eosinophils are associated with increased IgA subclass levels in the sinonasal mucosa of chronic rhinosinusitis with polyp patients
Authors: Aazami Hossein , Seif Farhad , Ghalehbaghi Babak , Babaheidarian Pegah , Mohebbi Alireza , Ahmadi Aslan , Khoshmirsafa Majid , Ghalehbaghi Sahand , Behnam Babak , Entezami Kobra Zinat , Madjd Zahra , Falak Reza
Published: April 2020

Background

Chronic rhinosinusitis (CRS) describes an inflammatory condition affecting the sinonasal mucosa. As the immune system players such as immunoglobulins play prominent roles in the development of CRS, we aimed to investigate the expression of IgA subclasses and factors involved in IgA class switching in the sinonasal mucosa of CRS patients.

Methods

Specimens were collected from the sinonasal mucosa of the healthy controls and CRS patients. Histological assessments were performed by H&E and immunohistochemistry. Real-time PCR and ELISA methods were applied to measure gene expression and protein levels extracted from tissue samples, respectively.

Results

We observed that total IgA and subclass-positive cells were higher in the patient groups than controls. There was a significant correlation between the number of eosinophils and total IgA and subclasses-positive cells (Pv < 0.0001). The expression of CXCL13, BAFF, AID, and germline transcripts were increased in CRSwNP patients. In contrast to IgA2 levels, IgA1 levels were significantly increased in the sinonasal tissue of CRSwNP patients (Pv < 0.01). TGF-β was significantly elevated in the sinonasal tissue of patients with CRSsNP.

Conclusions

Increased protein levels of IgA subclasses and related antibody-producing cells were associated with elevated eosinophils in CRSwNP patients which may result in eosinophil pathological functions. Several therapeutic approaches might be developed to modulate the IgA production to ameliorate the inflammatory mechanisms in CRSwNP patients.
Imaging diagnosis and staging of pancreatic ductal adenocarcinoma: a comprehensive review
Authors: Elbanna Khaled Y. , Jang Hyun-Jung , Kim Tae Kyoung
Published: April 2020

Pancreatic ductal adenocarcinoma (PDAC) has continued to have a poor prognosis for the last few decades in spite of recent advances in different imaging modalities mainly due to difficulty in early diagnosis and aggressive biological behavior. Early PDAC can be missed on CT due to similar attenuation relative to the normal pancreas, small size, or hidden location in the uncinate process. Tumor resectability and its contingency on the vascular invasion most commonly assessed with multi-phasic thin-slice CT is a continuously changing concept, particularly in the era of frequent neoadjuvant therapy. Coexistent celiac artery stenosis may affect the surgical plan in patients undergoing pancreaticoduodenectomy. In this review, we discuss the challenges related to the imaging of PDAC. These include radiological and clinical subtleties of the tumor, evolving imaging criteria for tumor resectability, preoperative diagnosis of accompanying celiac artery stenosis, and post-neoadjuvant therapy imaging. For each category, the key imaging features and potential pitfalls on cross-sectional imaging will be discussed. Also, we will describe the imaging discriminators of potential mimickers of PDAC.
Sex-specific clustering of metabolic risk factors and cancer risk: a longitudinal study in Iran
Authors: Ramezankhani Azra , Azizi Fereidoun , Hadaegh Farzad
Published: April 2020

Background

Cancer is a major cause of death in low- and middle-income countries. A large number of studies have shown that some of the metabolic risk factors (MRFs) tend to cluster in individuals. We examined the synergistic effects of multiple MRFs and cancer risk among Iranian adults.

Methods

Among 8593 (3929 men) participants aged ≥ 30 years, the self-organizing map (SOM) was applied to clustering of four MRFs including high fasting plasma glucose (HFPG), high total cholesterol (HTC), high systolic blood pressure (HSBP), and high body mass index (HBMI). The Cox proportional hazards model was used to investigate the association between clusters with cancer incidence during a median of 14.0 years of follow-up.

Results

During the study period, 265 new cases of cancer were identified among participants at risk. The incidence density rate was 2.5 per 1000 person years in total population. About 32 and 40% of men and women, respectively, had three or four MRFs. We identified seven clusters of MRFs in both men and women. In both genders, MRFs were clustered in those with older age. Further, inverse associations were found between current smoking in men, and education level and passive smoking in women and clustering of MRFs. In men, a cluster with 100% HSBP and HBMI had the highest risk for overall cancer. While, among women, a cluster with 100% HFPG and 93% HBMI yielded the highest risk for cancer. The risk was decreased when HBMI accompanied by HTC.

Conclusions

Clustering patterns may reflect underlying link between MRFs and cancer and could potentially facilitate tailored health promotion interventions.