scholarly journals Whole Transcriptome Analysis of Aedes albopictus Mosquito Head and Thorax Post-Chikungunya Virus Infection

Pathogens ◽  
2019 ◽  
Vol 8 (3) ◽  
pp. 132 ◽  
Author(s):  
Ravi kiran Vedururu ◽  
Matthew J. Neave ◽  
Vinod Sundaramoorthy ◽  
Diane Green ◽  
Jennifer A. Harper ◽  
...  
Keyword(s):  
Viral Infection ◽  
Aedes Albopictus ◽  
Post Infection ◽  
Chikungunya Virus ◽  
Quantitative Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Inhibitor ◽  
Significant Difference ◽  
Whole Transcriptome Analysis ◽  
Whole Transcriptome ◽  
Infectivity Titer

Chikungunya virus (CHIKV) is transmitted by Aedes mosquitoes and causes prolonged arthralgia in patients. After crossing the mosquito midgut barrier, the virus disseminates to tissues including the head and salivary glands. To better understand the interaction between Aedes albopictus and CHIKV, we performed RNASeq analysis on pools of mosquito heads and parts of the thorax 8 days post infection, which identified 159 differentially expressed transcripts in infected mosquitos compared to uninfected controls. After validation using RT-qPCR (reverse transcriptase-quantitative polymerase chain reaction), inhibitor of Bruton’s tyrosine kinase (BTKi), which has previously been shown to be anti-inflammatory in mammals after viral infection, was further evaluated for its functional significance. Knockdown of BTKi using double-stranded RNA in a mosquito cell line showed no significant difference in viral RNA or infectivity titer. However, BTKi gene knocked-down cells showed increased apoptosis 24 hours post-infection compared with control cells, suggesting involvement of BTKi in the mosquito response to viral infection. Since BTK in mammals promotes an inflammatory response and has been shown to be involved in osteoclastogenesis, a hallmark of CHIKV pathogenesis, our results suggest a possible conserved mechanism at play between mosquitoes and mammals. Taken together, these results will add to our understanding of Aedes Albopictus interactions with CHIKV.

scholarly journals RNASeq analysis ofAedes albopictusmosquitoes during chikungunya virus infection

2018 ◽  
Author(s):  
Ravikiran Vedururu ◽  
Matthew J. Neave ◽  
Mary Tachedjian ◽  
Paul R. Gorry ◽  
Jean-Bernard Duchemin ◽  
...  
Keyword(s):  
Differentially Expressed Genes ◽  
Aedes Albopictus ◽  
Post Infection ◽  
Viral Genome ◽  
Chikungunya Virus ◽  
Differentially Expressed ◽  
Single Amino Acid ◽  
The World ◽  
Whole Transcriptome ◽  
Mosquito Infection

AbstractChikungunya virus (CHIKV), preferentially transmitted byAedesmosquitoes, is an emerging pathogen around the world and causes significant morbidity in patients. A single amino acid mutation in the envelope protein of CHIKV has led to shift in vector preference towardsAedes albopictus, an invasive mosquito. Previous studies have shown that after infection, mosquitoes mount an antiviral immune response. However, molecular interactions during the course of infection at different tissues and time-points remain largely uncharacterised. Here we performed whole transcriptome analysis on dissected midguts and head/thorax of CHIKV (Indian Ocean strain) infectedAedes albopictusto identify differentially expressed genes compared with uninfected controls. For this, RNA was extracted at two days post-infection (D2) from pooled midguts and eight days post-infection (D8) from heads and the anterior 1/3rdof the thorax. We identified 25 and 96 differentially expressed genes from the D2 and D8 samples respectively (p-value <0.05). Customde novotranscriptomes were assembled for the reads that did not align with the reference genome and an additional 225 and 4771 differentially expressed genes from D2 and D8, respectively, were identified. Twenty-two of the identified transcripts, possibly involved in immunity, were validated by qRT-PCR. Interestingly, we also detected changes in viral diversity, as shown by number of mutations in the viral genome, with increase in number of mutations in the midgut compared with mammalian host (Vero cell culture), followed by reduction in the number of mutations in head and thorax at D8, indicating a possible genomic bottleneck. Taken together, these results will help in understanding AedesAlbopictusinteractions with CHIKV and can be utilised to reduce the impact of this viral infection.Author SummaryChikungunya virus has caused several outbreaks around the world in the last decade. Once a relatively unknown virus, it now causes seasonal infections in tropical and some temperate regions. This change in epidemiology is attributed to vector switch fromAedes aegyptitoAedes albopictus, an invasive pest leading to spread and causing infections in temperate regions. Although recent research has identified mosquito factors influencing infections, our understanding of interaction between chikungunya virus and its vector is limited. Using whole transcriptome sequencing of chikungunya infected mosquitoes, we identified differentially expressed genes in the midgut and head and thorax, over the course of mosquito infection. We also detected changes in the viral genome during mosquito infection and a possible genetic bottleneck event with reduction in viral variants at the head and thorax region of mosquito in the later stages of infection. These results will lead to improving our understanding of mosquito-virus interactions withAedes albopictusas a vector and in turn lead to development of novel disease control strategies.

scholarly journals Transcriptomic Profiling of Circulating Extrafollicular Helper T-like Cells in Patients with Active Chronic Graft-Versus-Host Disease Reveals Distinct Apoptosis Resistance

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 4882-4882
Author(s):  
Jiapei Liu ◽  
Kaibo Yang ◽  
Hua Jin ◽  
Qifa Liu
Keyword(s):  
Molecular Mechanism ◽  
Conflicts Of Interest ◽  
Human Peripheral Blood ◽  
Quantitative Polymerase Chain Reaction ◽  
Tumor Necrosis Factor Receptor ◽  
Cell Receptor ◽  
Apoptosis Resistance ◽  
Whole Transcriptome Analysis ◽  
Whole Transcriptome ◽  
Necrosis Factor

Abstract In our previous studies, we identified circulating extrafollicular helper T-like cells (CD44 hiCD62L loPSGL-1 loCD4 +, c-extrafollicular Th-like) in human peripheral blood. C-extrafollicular Th-like cells are associated with the development of cGVHD. However, the exact molecular mechanism of these cells in patient with active cGVHD is still unclear. We performed the whole transcriptome analysis of c-extrafollicular Th-like cells from patients with active cGVHD and without cGVHD to explore the molecular mechanism. We identified 4661 differentially expressed genes between two groups by RNA-sequencing. Upregulated expression of Ca2 + influx and protein kinase C signaling pathway induced genes that establish T cell receptor hyper-activation signature were observed in active cGVHD patients. Expression of several inflammation cytokines and receptors were also increased, including IL23, IL27, IL2RA, IL32, IL24 and IL7R. Furthermore, tumor necrosis factor receptor associated factor 1 (TRAF1) and Bcl-2 genes that linked to resist apoptosis were found upregulated. Consistently, we confirmed that the BCL-2 expression of c-extrafollicular helper Th-like cells was significant higher in active cGVHD patients than no cGVHD patients(P=0.02) by quantitative polymerase chain reaction (qPCR). Our study sheds new light on molecular mechanism of the c-extrafollicular Th-like in patient with active cGVHD. Targeting these signaling pathways might blunt the development of cGVHD. Disclosures No relevant conflicts of interest to declare.

scholarly journals Chikungunya Virus Transmission at Low Temperature by Aedes albopictus Mosquitoes

Pathogens ◽  
2019 ◽  
Vol 8 (3) ◽  
pp. 149 ◽  
Author(s):  
B. M. C. Randika Wimalasiri-Yapa ◽  
Liesel Stassen ◽  
Wenbiao Hu ◽  
Laith Yakob ◽  
Elizabeth A. McGraw ◽  
...  
Keyword(s):  
South African ◽  
Aedes Albopictus ◽  
Post Infection ◽  
Chikungunya Virus ◽  
Virus Transmission ◽  
Early Time ◽  
Vero Cells ◽  
Ambient Temperatures ◽  
Time Points ◽  
Central South

Aedes albopictus is an important vector of chikungunya virus (CHIKV). In Australia, Ae. albopictus is currently only known to be present on the islands of the Torres Strait but, should it invade the mainland, it is projected to spread to temperate regions. The ability of Australian Ae. albopictus to transmit CHIKV at the lower temperatures typical of temperate areas has not been assessed. Ae. albopictus mosquitoes were orally challenged with a CHIKV strain from either Asian or East/Central/South African (ECSA) genotypes (107 pfu/mL), and maintained at a constant temperature of either 18 °C or 28 °C. At 3- and 7-days post-infection (dpi), CHIKV RNA copies were quantified in mosquito bodies, and wings and legs using real time polymerase chain reaction (qRT-PCR), while the detection of virus in saliva (a proxy for transmission) was performed by amplification in cell culture followed by observation of cytopathic effect in Vero cells. Of the ≥95% of Ae. albopictus that survived to 7 dpi, all mosquitoes became infected and showed body dissemination of CHIKV at both temperatures and time points. Both the Asian and ECSA CHIKV genotypes were potentially transmissible by Australian Ae. albopictus at 28 °C within 3 days of oral challenge. In contrast, at 18 °C none of the mosquitoes showed evidence of ability to transmit either genotype of CHIKV at 3 dpi. Further, at 18 °C only Ae. albopictus infected with the ECSA genotype showed evidence of virus in saliva at 7 dpi. Overall, infection with the ECSA CHIKV genotype produced higher virus loads in mosquitoes compared to infection with the Asian CHIKV genotype. Our results suggest that lower ambient temperatures may impede transmission of some CHIKV strains by Ae. albopictus at early time points post infection.

scholarly journals RNASeq Analysis of Aedes albopictus Mosquito Midguts after Chikungunya Virus Infection

Viruses ◽  
2019 ◽  
Vol 11 (6) ◽  
pp. 513 ◽  
Author(s):  
Ravi kiran Vedururu ◽  
Matthew J. Neave ◽  
Mary Tachedjian ◽  
Melissa J. Klein ◽  
Paul R. Gorry ◽  
...  
Keyword(s):  
Differentially Expressed Genes ◽  
Aedes Albopictus ◽  
Post Infection ◽  
Chikungunya Virus ◽  
De Novo ◽  
Differentially Expressed ◽  
Single Amino Acid ◽  
P Value ◽  
Antiviral Immune Response ◽  
The Impact

Chikungunya virus (CHIKV) is an emerging pathogen around the world and causes significant morbidity in patients. A single amino acid mutation in the envelope protein of CHIKV has led to a shift in vector preference towards Aedes albopictus. While mosquitoes are known to mount an antiviral immune response post-infection, molecular interactions during the course of infection at the tissue level remain largely uncharacterised. We performed whole transcriptome analysis on dissected midguts of Aedes albopictus infected with CHIKV to identify differentially expressed genes. For this, RNA was extracted at two days post-infection (2-dpi) from pooled midguts. We initially identified 25 differentially expressed genes (p-value < 0.05) when mapped to a reference transcriptome. Further, multiple differentially expressed genes were identified from a custom de novo transcriptome, which was assembled using the reads that did not align with the reference genome. Thirteen of the identified transcripts, possibly involved in immunity, were validated by qRT-PCR. Homologues of seven of these genes were also found to be significantly upregulated in Aedes aegypti midguts 2 dpi, indicating a conserved mechanism at play. These results will help us to characterise the molecular interaction between Aedes albopictus and CHIKV and can be utilised to reduce the impact of this viral infection.

scholarly journals Whole Transcriptome Analysis Revealed a Stress Response to Deep Underground Environment Conditions in Chinese Hamster V79 Lung Fibroblast Cells

2021 ◽  
Vol 12 ◽  
Author(s):  
Liju Duan ◽  
Hongying Jiang ◽  
Jifeng Liu ◽  
Yilin Liu ◽  
Tengfei Ma ◽  
...  
Keyword(s):  
Transcriptome Analysis ◽  
Background Radiation ◽  
Lung Fibroblast ◽  
Fibroblast Cells ◽  
V79 Cells ◽  
Significant Difference ◽  
Deep Underground ◽  
Whole Transcriptome Analysis ◽  
Whole Transcriptome ◽  
Cells Cultured

Background: Prior studies have shown that the proliferation of V79 lung fibroblast cells could be inhibited by low background radiation (LBR) in deep underground laboratory (DUGL). In the current study, we revealed further molecular changes by performing whole transcriptome analysis on the expression profiles of long non-coding RNA (lncRNA), messenger RNA (mRNA), circular RNA (circRNA) and microRNA (miRNA) in V79 cells cultured for two days in a DUGL.Methods: Whole transcriptome analysis including lncRNA, mRNAs, circ RNA and miRNA was performed in V79 cells cultured for two days in DUGL and above ground laboratory (AGL), respectively. The differentially expressed (DE) lncRNA, mRNA, circRNA, and miRNA in V79 cells were identified by the comparison between DUGL and AGL groups. Quantitative real-time polymerase chain reaction(qRT-PCR)was conducted to verify the selected RNA sequencings. Then, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway was analyzed for the DE mRNAs which enabled to predict target genes of lncRNA and host genes of circRNA.Results: With |log2(Fold-change)| ≥ 1.0 and p &lt; 0.05, a total of 1257 mRNAs (353 mRNAs up-regulated, 904 mRNAs down-regulated), 866 lncRNAs (145 lncRNAs up-regulated, 721 lncRNAs down-regulated), and 474 circRNAs (247 circRNAs up-regulated, 227 circRNAs down-regulated) were significantly altered between the two groups. There was no significant difference in miRNA between the two groups. The altered RNA profiles were mainly discovered in lncRNAs, mRNAs and circRNAs. DE RNAs were involved in many pathways including ECM-RI, PI3K-Akt signaling, RNA transport and the cell cycle under the LBR stress of the deep underground environment.Conclusion: Taken together, these results suggest that the LBR in the DUGL could induce transcriptional repression, thus reducing metabolic process and reprogramming the overall gene expression profile in V79 cells.

The Fine Structure of Replicating Simian Adenoviruses in LLC-MK2

1968 ◽  
Vol 26 ◽  
pp. 220-221
Author(s):  
Matias Pardo ◽  
Malcolm Slifkin ◽  
Leonard Merkow ◽  
Marie Sanchez
Keyword(s):  
Tissue Culture ◽  
Post Infection ◽  
Cesium Chloride ◽  
Longitudinal Section ◽  
Tubular Structures ◽  
Virus Particles ◽  
Ultrastructural Studies ◽  
Culture Cells ◽  
Simian Adenovirus ◽  
Infectivity Titer

The simian adenoviruses SV20, SV30 and SA7 have been found to be oncogenic in the Syrian hamster. The growth characteristics and replicative cycle of these viruses in tissue culture therefore appeared appropriate to investigate. Cesium chloride purified simian adenovirus with an infectivity titer of 100 TCID50, was inoculated into monolayers of LLC-MK2 cells. Cells were fixed in osmium tetroxide and embedded for ultrastructural studies at 1, 3, 6, 9, 18, 24, 48, 72, 120 and 192 hours post-infection.At the first hour post-infection, virus particles were adsorbed to the plasmalemma and found within the peripheral cytoplasm of many LLC-MK2 cells (Fig. 1). Although the first detection of infectious virus occurred at 14 hours and infectivity titers did not reach a maximum until 30 hours, intranuclear virus particles were observed by 3 hours in typical adenovirus crystalline array (Fig. 2) by means of electron microscopy. These typical honeycomb arrayed virus particles at 3 hours provided evidence of significant replication in approximately 5 percent of tissue culture cells examined. Simultaneously, a classical nuclear inclusion manifested by peripheral condensation of nuclear chromatin was evident by light microscopy. As early at 6 to 9 hours, unusual intranuclear concentric membranes formed “tubes” which contained linear arranged virus particles (Fig. 3). In transverse or tangential sections, these “tubes” appeared cochlear-like in shape. In longitudinal section, these intranuclear tubular structures contained individual virus particles at various stages of maturation in a linear arranged order. This arrangement resembled “peas in a pod”.

Gene expression analysis in modeling Parkinson’s disease

2020 ◽  
pp. 103-104
Author(s):  
М.М. Руденок ◽  
А.Х. Алиева ◽  
А.А. Колачева ◽  
М.В. Угрюмов ◽  
П.А. Сломинский ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Early Stage ◽  
Systemic Disease ◽  
Molecular Genetic ◽  
Presymptomatic Stage ◽  
Whole Transcriptome Analysis ◽  
Whole Transcriptome ◽  
The Brain ◽  
Transcriptome Level

Несмотря на очевидный прогресс, достигнутый в изучении молекулярно-генетических факторов и механизмов патогенеза болезни Паркинсона (БП), в настоящее время стало ясно, что нарушения в структуре ДНК не описывают весь спектр патологических изменений, наблюдаемых при развитии заболевания. В настоящее время показано, что существенное влияние на патогенез БП могут оказывать изменения на уровне транскриптома. В работе были использованы мышиные модели досимптомной стадии БП, поздней досимптомной и ранней симптомной (РСС) стадиями БП. Для полнотранскриптомного анализа пулов РНК тканей черной субстанции и стриатума мозга мышей использовались микрочипы MouseRef-8 v2.0 Expression BeadChip Kit («Illumina», США). Полученные данные указывают на последовательное вовлечение транскриптома в патогенез БП, а также на то, что изменения на транскриптомном уровне процессов транспорта и митохондриального биогенеза могут играть важную роль в нейродегенерации при БП уже на самых ранних этапах. Parkinson’s disease (PD) is a complex systemic disease, mainly associated with the death of dopaminergic neurons. Despite the obvious progress made in the study of molecular genetic factors and mechanisms of PD pathogenesis, it has now become clear that violations in the DNA structure do not describe the entire spectrum of pathological changes observed during the development of the disease. It has now been shown that changes at the transcriptome level can have a significant effect on the pathogenesis of PD. The authors used models of the presymptomatic stage of PD with mice decapitation after 6 hours (6 h-PSS), presymptomatic stage with decapitation after 24 hours (24 h-PSS), advanced presymptomatic (Adv-PSS) and early symptomatic (ESS) stages of PD. For whole transcriptome analysis of RNA pools of the substantia nigra and mouse striatum, the MouseRef-8 v2.0 Expression BeadChip Kit microchips (Illumina, USA) were used. As a result of the analysis of whole transcriptome data, it was shown that, there are a greater number of statistically significant changes in the tissues of the brain and peripheral blood of mice with Adv-PSS and ESS models of PD compared to 6 h-PSS and 24 h-PSS models. In general, the obtained data indicate the sequential involvement of the transcriptome in the pathogenesis of PD, as well as the fact that changes at the transcriptome level of the processes of transport and mitochondrial biogenesis can play an important role in neurodegeneration in PD at an early stage.

scholarly journals Epidemiological characteristics of four common respiratory viral infections in children

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Guohong Zhu ◽  
Dan Xu ◽  
Yuanyuan Zhang ◽  
Tianlin Wang ◽  
Lingyan Zhang ◽  
...  
Keyword(s):  
Respiratory Tract ◽  
Viral Infection ◽  
Influenza A ◽  
Multiple Infection ◽  
Influenza B ◽  
Preschool Period ◽  
Significant Difference ◽  
Different Seasons ◽  
Tract Infections ◽  
Epidemiological Characteristics

Abstract Background Viruses are the main infectious agents of acute respiratory infections in children. We aim to describe the epidemiological characteristics of viral pathogens of acute respiratory tract infections in outpatient children. Methods From April 2018 to March 2019, the results of viral detection using oral pharyngeal swabs from 103,210 children with acute respiratory tract infection in the outpatient department of the Children’s Hospital, Zhejiang University School of Medicine, were retrospectively analyzed. Viral antigens, including adenovirus (ADV), influenza A (FLUA), influenza B (FLUB) and respiratory syncytial virus (RSV), were detected by the colloidal gold method. Results At least one virus was detected in 38,355 cases; the positivity rate was 37.2%. A total of 1910 cases of mixed infection with two or more viruses were detected, and the positivity rate of multiple infection was 1.9%. The ADV positivity rate was highest in the 3–6-year-old group (18.7%), the FLUA positivity rate was highest in the > 6-year-old group (21.6%), the FLUB positivity rate was highest in the > 6-year-old group (6.6%), and the RSV positivity rate was highest in the < 1-year-old group (10.6%). There was a significant difference in the positivity rate of viral infection among different age groups (χ2 = 1280.7, P < 0.001). The rate of positive viral infection was highest in winter (47.1%). The ADV infection rate was highest in spring (18.2%). The rates of FLUA and FLUB positivity were highest in winter (28.8% and 3.6%, respectively). The rate of RSV positivity was highest in autumn (17.4%). The rate of positive viral infection in different seasons was significantly different (χ2 = 6459.1, P < 0.001). Conclusions Viral infection rates in children differ for different ages and seasons. The positivity rate of ADV is highest in the preschool period and that of RSV is highest in infants; that of FLU increases with age. The total positive rate of viral infection in different seasons is highest in winter, as is the rate of FLU positivity.

scholarly journals Implementation of a deep learning model for automated classification of Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse) in real time

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Song-Quan Ong ◽  
Hamdan Ahmad ◽  
Gomesh Nair ◽  
Pradeep Isawasan ◽  
Abdul Hafiz Ab Majid
Keyword(s):  
Deep Learning ◽  
Aedes Aegypti ◽  
Real Time ◽  
Aedes Albopictus ◽  
Automated Classification ◽  
Expert Performance ◽  
Deep Convolutional Neural Networks ◽  
Significant Difference ◽  
Set Up

AbstractClassification of Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse) by humans remains challenging. We proposed a highly accessible method to develop a deep learning (DL) model and implement the model for mosquito image classification by using hardware that could regulate the development process. In particular, we constructed a dataset with 4120 images of Aedes mosquitoes that were older than 12 days old and had common morphological features that disappeared, and we illustrated how to set up supervised deep convolutional neural networks (DCNNs) with hyperparameter adjustment. The model application was first conducted by deploying the model externally in real time on three different generations of mosquitoes, and the accuracy was compared with human expert performance. Our results showed that both the learning rate and epochs significantly affected the accuracy, and the best-performing hyperparameters achieved an accuracy of more than 98% at classifying mosquitoes, which showed no significant difference from human-level performance. We demonstrated the feasibility of the method to construct a model with the DCNN when deployed externally on mosquitoes in real time.

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