scholarly journals Symptom Severity, Infection Progression and Plant Responses in Solanum Plants Caused by Three Pospiviroids Vary with the Inoculation Procedure

2021 ◽  
Vol 22 (12) ◽  
pp. 6189
Author(s):  
Francisco Vázquez Prol ◽  
Joan Márquez-Molins ◽  
Ismael Rodrigo ◽  
María Pilar López-Gresa ◽  
José María Bellés ◽  
...  
Keyword(s):  
Ribosome Biogenesis ◽  
Solanum Melongena ◽  
Potato Spindle Tuber Viroid ◽  
Signal Molecules ◽  
Plant Responses ◽  
Host Interactions ◽  
Tomato Chlorotic Dwarf Viroid ◽  
Ribosomal Stress

Infectious viroid clones consist of dimeric cDNAs used to generate transcripts which mimic the longer-than-unit replication intermediates. These transcripts can be either generated in vitro or produced in vivo by agro-inoculation. We have designed a new plasmid, which allows both inoculation methods, and we have compared them by infecting Solanum lycopersicum and Solanum melongena with clones of Citrus exocortis virod (CEVd), Tomato chlorotic dwarf viroid (TCDVd), and Potato spindle tuber viroid (PSTVd). Our results showed more uniform and severe symptoms in agro-inoculated plants. Viroid accumulation and the proportion of circular and linear forms were different depending on the host and the inoculation method and did not correlate with the symptoms, which correlated with an increase in PR1 induction, accumulation of the defensive signal molecules salicylic (SA) and gentisic (GA) acids, and ribosomal stress in tomato plants. The alteration in ribosome biogenesis was evidenced by both the upregulation of the tomato ribosomal stress marker SlNAC082 and the impairment in 18S rRNA processing, pointing out ribosomal stress as a novel signature of the pathogenesis of nuclear-replicating viroids. In conclusion, this updated binary vector has turned out to be an efficient and reproducible method that will facilitate the studies of viroid–host interactions.

scholarly journals Formation of a Complex between Nucleolin and Replication Protein a after Cell Stress Prevents Initiation of DNA Replication

2000 ◽  
Vol 149 (4) ◽  
pp. 799-810 ◽  
Author(s):  
Yaron Daniely ◽  
James A. Borowiec
Keyword(s):  
Dna Replication ◽  
Ribosome Biogenesis ◽  
Protein A ◽  
Simian Virus 40 ◽  
Replication Protein A ◽  
Cell Stress ◽  
Replication Protein ◽  
Binding Studies

We used a biochemical screen to identify nucleolin, a key factor in ribosome biogenesis, as a high-affinity binding partner for the heterotrimeric human replication protein A (hRPA). Binding studies in vitro demonstrated that the two proteins physically interact, with nucleolin using an unusual contact with the small hRPA subunit. Nucleolin significantly inhibited both simian virus 40 (SV-40) origin unwinding and SV-40 DNA replication in vitro, likely by nucleolin preventing hRPA from productive interaction with the SV-40 initiation complex. In vivo, use of epifluorescence and confocal microscopy showed that heat shock caused a dramatic redistribution of nucleolin from the nucleolus to the nucleoplasm. Nucleolin relocalization was concomitant with a tenfold increase in nucleolin–hRPA complex formation. The relocalized nucleolin significantly overlapped with the position of hRPA, but only poorly with sites of ongoing DNA synthesis. We suggest that the induced nucleolin–hRPA interaction signifies a novel mechanism that represses chromosomal replication after cell stress.

scholarly journals Osteoclast-Derived Extracellular miR-106a-5p Promotes Osteogenic Differentiation and Facilitates Bone Defect Healing

2021 ◽  
Author(s):  
Yutong Wu ◽  
Hongbo Ai ◽  
Yuchi Zou ◽  
Jianzhong Xu
Keyword(s):  
Demineralized Bone Matrix ◽  
Bone Matrix ◽  
Signal Molecules ◽  
Calvarial Defect ◽  
Communication Mode ◽  
Defect Healing ◽  
Bone Defect Healing ◽  
Intercellular Communications

Abstract Small extracellular vesicles (sEVs) are considered to play critical roles in intercellular communications during normal and pathological processes since they are enriched with miRNAs and other signal molecules. In bone remodeling, osteoclasts generate large amounts of sEVs. However, there is very little research about whether and how osteoclast-derived sEVs (OC-sEVs) affect surrounding cells. In our study, microarray analysis identified miR-106a-5p highly enriched in OC-sEV. Further experiments confirmed that OC-sEVs inhibited Fam134a through miR-106a-5p and significantly promoted bone mesenchymal stem cell (BMSC) osteogenic mineralization in vitro. Next, we prepared sEV-modified demineralized bone matrix (DBM) as a repair scaffold, and used a calvarial defect mouse model to evaluate the pro-osteogenic activities of the scaffold. In vivo result indicated DBM modified with miR-106a-5p-sEVs showed an enhanced capacity of bone regeneration. This important finding further emphasizes that sEV-mediated miR-106a-5p transfer play critical roles in osteogenesis and indicate a novel communication mode between osteoclasts and BMSCs.

scholarly journals Good and Bad Neighbourhoods in Viral Sequence Space: Predicting, Altering, Targeting Virus Populations

Proceedings ◽  
2020 ◽  
Vol 50 (1) ◽  
pp. 61
Author(s):  
Marco Vignuzzi
Keyword(s):  
Sequence Space ◽  
Experimental Evolution ◽  
Point Mutations ◽  
Amino Acid Sequences ◽  
Viral Population ◽  
Genome Composition ◽  
In Vivo Models ◽  
Host Interactions

All viruses, but especially RNA viruses, generate tremendous diversity in genome composition, including point mutations, duplications, deletions, and insertions. We used in vitro and in vivo models to perform natural and directed experimental evolution. We then combined the resulting data with mathematical modelling to determine how virus populations occupy sequence space—a multidimensional hypercube that describes all combinations of nucleotide, codon, or amino acid sequences. In this study, we demonstrate how these experimental and computational approaches can help monitor, predict, alter, and even target virus evolution and population dynamics, creating new ways to study virus–host interactions and to innovate antiviral approaches. Using arboviruses, enteroviruses, and influenza, we recreate and predict host jumps and emergence events in the lab, redirect evolution towards the ‘bad’ neighbourhoods of sequence space that represent attenuation, and poison the viral population by disturbing the balance between good and bad genomes.

Corilagin ameliorates schistosomiasis hepatic fibrosis through regulating IL-13 associated signal pathway in vitro and in vivo

Parasitology ◽  
2016 ◽  
Vol 143 (12) ◽  
pp. 1629-1638 ◽  
Author(s):  
HUA-RONG LI ◽  
GANG LI ◽  
MAN LI ◽  
SHU-LING ZHANG ◽  
HENG WANG ◽  
...  
Keyword(s):  
Hepatic Fibrosis ◽  
Quantitative Polymerase Chain Reaction ◽  
Signal Pathway ◽  
Signal Molecules ◽  
Mrna Levels ◽  
Protein Levels ◽  
Set Up ◽  
Inflammatory Changes

SUMMARYInterleukin (IL)-13-associated signal pathway plays an important role in schistosomiasis hepatic fibrosis. In this study we tried to investigate the effects of corilagin to ameliorate schistosomiasis hepatic fibrosis through regulating IL-13-associated signal pathway in vitro and in vivo. Cellular model was set up with hepatic stellate cells-T6 cells stimulated by rIL-13 and male Balb/c mice were infected with Schistosoma japonicum cercariaeas as animal model. Liver histological changes were observed with haematoxylin and eosin staining. Masson staining was employed to observe the change of egg granulomas. Expression of Col (collagen) and Col III were examined with Immunohistochemistry. Western bolt was employed to detect the JAK-1 and IL13Rα1 proteins. The mRNA expression of Col I, Col III, IL-13, JAK-1 and IL13Rα1 were tested by quantitative polymerase chain reaction. As a result, less inflammatory changes were found in all corilagin groups compared with model group and praziquantel group. The mRNA levels of Col I, Col III, IL-13, JAK-1 and IL13Rα1 were significantly decreased after corilagin intervention (P < 0·01). JAK-1 and IL-13Rα1 protein levels were also greatly decreased in the corilagin groups (P < 0·01). In conclusion, corilagin could ameliorate schistosomiasis hepatic fibrosis by down-regulating the expression of IL-13 and signal molecules in IL-13 pathway.

scholarly journals Adaptive Strategies and Pathogenesis of Clostridium difficile fromIn VivoTranscriptomics

2013 ◽  
Vol 81 (10) ◽  
pp. 3757-3769 ◽  
Author(s):  
Claire Janoir ◽  
Cécile Denève ◽  
Sylvie Bouttier ◽  
Frédéric Barbut ◽  
Sandra Hoys ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Stress Responses ◽  
Differentially Expressed ◽  
Content Type ◽  
Host Interactions ◽  
Colonization Factor ◽  
Genes Encoding ◽  
Colonization Process

ABSTRACTClostridium difficileis currently the major cause of nosocomial intestinal diseases associated with antibiotic therapy in adults. In order to improve our knowledge ofC. difficile-host interactions, we analyzed the genome-wide temporal expression ofC. difficile630 genes during the first 38 h of mouse colonization to identify genes whose expression is modulatedin vivo, suggesting that they may play a role in facilitating the colonization process. In the ceca of theC. difficile-monoassociated mice, 549 genes of theC. difficilegenome were differentially expressed compared to their expression duringin vitrogrowth, and they were distributed in several functional categories. Overall, our results emphasize the roles of genes involved in host adaptation. Colonization results in a metabolic shift, with genes responsible for the fermentation as well as several other metabolic pathways being regulated inversely to those involved in carbon metabolism. In addition, several genes involved in stress responses, such as ferrous iron uptake or the response to oxidative stress, were regulatedin vivo. Interestingly, many genes encoding conserved hypothetical proteins (CHP) were highly and specifically upregulatedin vivo. Moreover, genes for all stages of sporulation were quickly inducedin vivo, highlighting the observation that sporulation is central to the persistence ofC. difficilein the gut and to its ability to spread in the environment. Finally, we inactivated two genes that were differentially expressedin vivoand evaluated the relative colonization fitness of the wild-type and mutant strains in coinfection experiments. We identified a CHP as a putative colonization factor, supporting the suggestion that thein vivotranscriptomic approach can unravel newC. difficilevirulence genes.

scholarly journals Origanum dictamnusOil Vapour Suppresses the Development of Grey Mould in Eggplant FruitIn Vitro

2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
Andriana Stavropoulou ◽  
Kostas Loulakakis ◽  
Naresh Magan ◽  
Nikos Tzortzakis
Keyword(s):  
Fruit Quality ◽  
Solanum Melongena ◽  
Grey Mould ◽  
Colony Growth ◽  
Disease Spread ◽  
Continuous Exposure ◽  
Conidial Production ◽  
Lesion Growth

Grey mould rot (Botrytis cinerea) developmentin vitroor in eggplant (Solanum melongenaL.) fruit was evaluated after treatment with dittany (Origanum dictamnusL.) oil (DIT) and storage at 12°C and 95% relative humidity during or following exposure to the volatiles. DIT volatiles used in different concentration (0-50-100-250 μL/L) and times of exposure (up to 120 h) examined the effects on pathogen development as well as fruit quality parameters.In vitro,fungal colony growth was inhibited with the application of DIT oil (during or after exposure) and/or time of application. Continuous exposure to oils reduced conidial germination and production with fungistatic effects observed in 250 μL/L.In vivo,fungal lesion growth and conidial production reduced in DIT-treated fruits. Interesting, in fruits preexposed to volatiles before fungal inoculation, DIT application induced fruit resistance against the pathogen, by reduced lesion growth and conidial production. Conidial viability reduced in >100 μL/L DIT oil. Fruits exposed to essential oil did not affect fruit quality related attributes in general, while skin lightness (Lvalue) increased in 50 and 100 μL/L DIT oil. The results of the current study indicated that dittany volatiles may be considered as an alternative food preservative, eliminating disease spread in the storage/transit atmospheres.

scholarly journals Bioatividade de óleos essenciais e extratos vegetais no controle de doenças causadas por Phytophthora nicotianae em solanáceas

2020 ◽  
Vol 46 (3) ◽  
pp. 267-272
Author(s):  
Antônio Alves Pimenta Neto ◽  
Gláucio Dias Gonçalves ◽  
Carolina Santos Benjamin ◽  
Larissa Corrêa do Bomfim Costa ◽  
Rosilene Aparecida de Oliveira ◽  
...  
Keyword(s):  
Lycopersicon Esculentum ◽  
Solanum Melongena ◽  
Phytophthora Nicotianae ◽  
Syzygium Aromaticum ◽  
Cymbopogon Nardus

RESUMO Óleos essenciais (EO) e extratos brutos aquosos (EBA) de Syzygium aromaticum e Cymbopogon nardus, e seus principais componentes químicos foram investigados para controle in vitro e in vivo de doenças causadas por Phytophthora nicotianae em tomateiro (Lycopersicon esculentum) e berinjela (Solanum melongena). Experimentos in vitro foram conduzidos pelo método de diluição em ágar com diferentes concentrações de óleos essenciais (OE’s) (0,1; 0,5; 1,0 ?L/mL) e extratos aquosos brutos (EBA’s) (1,0; 10,0 20,0%) para avaliar o potencial inibitório sobre o crescimento micelial e germinação de zoósporos. Com as porcentagens de inibição e concentrações inibitórias mínimas encontradas, realizou-se testes em frutos e plântulas sob ambiente controlado. As variáveis avaliadas foram o diâmetro médio das lesões formadas na superfície dos frutos e a incidência e morte de plântulas ao longo de seis e 15 dias de avaliação, respectivamente. Os produtos que mais inibiram o crescimento micelial e a germinação de zoósporos foram obtidos de S. aromaticum, nas concentrações de 0,5 ?L/mL e 10% de OE e EBA, respectivamente. Os tratamentos que mais retardaram a progressão da doença em frutos e plântulas, em comparação com o controle, foram o OE e EBA de C. nardus a 1,0 ?L/mL e 20%, respectivamente. Portanto, os produtos obtidos de S. aromaticum e C. nardus, têm potencial para reduzir o ataque deste patógeno em tomate e berinjela.

scholarly journals Extracellular vesicles (exosomes) in prokaryotic organisms: role in their biology and realization of their pathogen potential

2020 ◽  
pp. 118-130
Author(s):  
B. A. Shenderov ◽  
A. B. Sinitsa ◽  
M. M. Zakharchenko ◽  
E. I. Tkachenko
Keyword(s):  
Recipient Cell ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Host Cells ◽  
Signal Molecules ◽  
Gram Positive Bacteria ◽  
Physiological Processes ◽  
Pathological Conditions

An increasing number of gram-negative and gram-positive bacteria have been observed to secrete outer- membrane vesicles (OMVs) during their growth both under physiological and pathological conditions in vitro and in vivo. These cell-derived particles are present in many — if not all — physiological fluids. They can convey the multiple various low weight effector and signal molecules (proteins, nucleic acids, lipids, and carbohydrates) into the bacterial and host cells that have important functions in their intercellular communication and regulation. Involvement of OMVS in the various biological functions of prokariotic and eukaryotic cells make them to be key players in both physiological processes and also in pathological conditions. Additionally, the ability of OMVs to deliver molecules to recipient cell opens the possibility of their use as novel disease biomarkers and as promising drug/therapy agents. In this Review, we describe the mechanisms through which bacterial OMVs can support the host homeostasis and health and induce host pathology or immune tolerance, and discuss the possibility of these OMVs participate in innovative nanobiotechnologies.

scholarly journals Divergent replication kinetics of two phenotypically different parvoviruses of rats

2001 ◽  
Vol 82 (3) ◽  
pp. 537-546 ◽  
Author(s):  
Lisa J. Ball-Goodrich ◽  
Elizabeth Johnson ◽  
Robert Jacoby
Keyword(s):  
Infectious Agent ◽  
Protein Accumulation ◽  
Sequence Comparisons ◽  
Minute Virus Of Mice ◽  
Phenotypic Differences ◽  
Host Interactions ◽  
Laboratory Rats ◽  
Kinetics Of

Rat virus (RV) is an important infectious agent of laboratory rats because of its high prevalence and capacity to disrupt research. Additionally, RV infection serves as a model for characterizing virus–host interactions during acute, persistent and prenatal infection. Our research has examined the pathogenesis of two RV strains, RV-UMass and RV-Y. RV-UMass is more pathogenic, causes a higher level of persistent infection and transmits to the foetus after oronasal inoculation of the pregnant dam. To determine in vitro distinctions between the strains that may account for these differences and to provide a benchmark for characterizing virus replication in vivo, synchronized in vitro replication of both RV strains was defined and compared. The results demonstrated that RV replication has replicative intermediates, virus transcripts and proteins similar to those reported for the prototype parvovirus, minute virus of mice. However, the replicative cycle of RV-UMass was 12 h compared with 24 h for RV-Y, and RV-UMass and RV-Y differed in kinetics of virus DNA replication, transcription and protein accumulation. Additionally, in situ analysis correlated well with kinetics data as determined by Southern and Northern blot analysis. Sequence comparisons between the strains also determined coding differences that may contribute to phenotypic differences.

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