scholarly journals Protein modification with ISG15 blocks coxsackievirus pathology by antiviral and metabolic reprogramming

2020 ◽  
Vol 6 (11) ◽  
pp. eaay1109 ◽  
Author(s):  
Meike Kespohl ◽  
Clara Bredow ◽  
Karin Klingel ◽  
Martin Voß ◽  
Anna Paeschke ◽  
...  
Keyword(s):  
Protein Modification ◽  
Shotgun Proteomics ◽  
Oxidative Capacity ◽  
Metabolic Reprogramming ◽  
Major Type ◽  
Infection Model ◽  
Type I ◽  
Synergistic Activity ◽  
Cardiac Damage ◽  
Species Specific

Protein modification with ISG15 (ISGylation) represents a major type I IFN–induced antimicrobial system. Common mechanisms of action and species-specific aspects of ISGylation, however, are still ill defined and controversial. We used a multiphasic coxsackievirus B3 (CV) infection model with a first wave resulting in hepatic injury of the liver, followed by a second wave culminating in cardiac damage. This study shows that ISGylation sets nonhematopoietic cells into a resistant state, being indispensable for CV control, which is accomplished by synergistic activity of ISG15 on antiviral IFIT1/3 proteins. Concurrent with altered energy demands, ISG15 also adapts liver metabolism during infection. Shotgun proteomics, in combination with metabolic network modeling, revealed that ISG15 increases the oxidative capacity and promotes gluconeogenesis in liver cells. Cells lacking the activity of the ISG15-specific protease USP18 exhibit increased resistance to clinically relevant CV strains, therefore suggesting that stabilizing ISGylation by inhibiting USP18 could be exploited for CV-associated human pathologies.

Repurposing of auranofin against bacterial infections: An In silico and In vitro study

2020 ◽  
Vol 16 ◽  
Author(s):  
Nidhi Sharma ◽  
Arti Singh ◽  
Ruchika Sharma ◽  
Anoop Kumar
Keyword(s):  
Broad Spectrum ◽  
In Silico ◽  
Antibacterial Agent ◽  
Bacterial Infections ◽  
In Vitro Assays ◽  
Infection Model ◽  
Synergistic Activity ◽  
Bacterial Strains ◽  
Molecular Docking Study

Aim: The aim of the study was to find out the role of auranofin as a promising broad spectrum antibacterial agent. Methods: In-vitro assays (Percentage growth retardation, Bacterial growth kinetics, Biofilm formation assay) and In-silico study (Molegro virtual docker (MVD) version 6.0 and Molecular operating environment (MOE) version 2008.10 software). Results: The in vitro assays have shown that auranofin has good antibacterial activity against Gram positive and Gram negative bacterial strains. Further, auranofin has shown synergistic activity in combination with ampicillin against S. aureus and B. subtilis whereas in combination with neomycin has just shown additive effect against E. coli, P. aeruginosa and B. pumilus. In vivo results have revealed that auranofin alone and in combination with standard drugs significantly decreased the bioburden in zebrafish infection model as compared to control. The molecular docking study have shown good interaction of auranofin with penicillin binding protein (2Y2M), topoisomerase (3TTZ), UDP-3-O-[3- hydroxymyristoyl] N-acetylglucosaminedeacetylase (3UHM), cell adhesion protein (4QRK), β-lactamase (5CTN) and arylsulphatase (1HDH) enzyme as that of reference ligand which indicate multimodal mechanism of action of auranofin. Finally, MTT assay has shown non-cytotoxic effect of auranofin. Conclusion: In conclusion, auranofin in combination with existing antibiotics could be developed as a broad spectrum antibacterial agent; however, further studies are required to confirm its safety and efficacy. This study provides possibility of use of auranofin apart from its established therapeutic indication in combination with existing antibiotics to tackle the problem of resistance.

Metabolic derangements in COPD muscle dysfunction

2013 ◽  
Vol 114 (9) ◽  
pp. 1282-1290 ◽  
Author(s):  
Luis Puente-Maestu ◽  
Alberto Lázaro ◽  
Blanca Humanes
Keyword(s):  
Lactate Production ◽  
Respiratory Muscles ◽  
Oxidative Capacity ◽  
Chronic Obstructive ◽  
Diabetic Patients ◽  
Peroxisome Proliferator ◽  
Type I ◽  
Oxidase Gene ◽  
Altered Expression ◽  
Copd Patients

Mitochondrial muscle alterations are common in patients with chronic obstructive pulmonary disease (COPD) and manifest mainly as decreased oxidative capacity and excessive production of reactive oxygen species (ROS). The significant loss of oxidative capacity observed in the quadriceps of COPD patients is mainly due to reduced mitochondrial content in the fibers, a finding consistent with the characteristic loss of type I fibers observed in that muscle. Decreased oxidative capacity does not directly limit maximum performance; however, it is associated with increased lactate production at lower exercise intensity and reduced endurance. Since type I fiber atrophy does not occur in respiratory muscles, the loss of such fibers in the quadriceps could be to the result of disuse. In contrast, excessive production of ROS and oxidative stress are observed in both the respiratory muscles and the quadriceps of COPD patients. The causes of increased ROS production are not clear, and a number of different mechanisms can play a role. Several mitochondrial alterations in the quadriceps of COPD patients are similar to those observed in diabetic patients, thus suggesting a role for muscle alterations in this comorbidity. Amino acid metabolism is also altered. Expression of peroxisome proliferator-activated receptor-γ coactivator-1α mRNA is low in the quadriceps of COPD patients, which could also be a consequence of type I fiber loss; nevertheless, its response to exercise is not altered. Patterns of muscle cytochrome oxidase gene activation after training differ between COPD patients and healthy subjects, and the profile is consistent with hypoxic stress, even in nonhypoxic patients.

Changes in the transferrin requirement of cultured chick embryo mesoderm cells during early differentiation

Development ◽  
1986 ◽  
Vol 95 (1) ◽  
pp. 81-93
Author(s):  
E. J. Sanders
Keyword(s):  
Chick Embryo ◽  
Type I Collagen ◽  
Specific Effect ◽  
Primitive Streak ◽  
Cellular Adhesion ◽  
Type I ◽  
Surface Binding ◽  
Surface Receptors ◽  
Early Differentiation ◽  
Species Specific

Mesodermal tissue from the chick embryo at various stages of early differentiation was cultured in hydrated gels of type I collagen in the presence and absence of transferrin. Primary mesoderm explants from primitive-streak-stage embryos responded to the presence of avian transferrin by significantly improved outgrowth which appeared to be related to the ability of the cells to attach to, and migrate in, the collagen. No evidence was obtained which suggested that this observation was dependent on increased cell proliferation. This outgrowth enhancement was not duplicated by transferrin of human origin. The avian transferrin did not produce this effect on cells cultured on plastic substrata, suggesting that the species-specific effect involves modulation by the extracellular matrix. Mesoderm explants from somite stages of development showed no increase in outgrowth in the presence of either avian or human transferrin as judged by counting the number of outwandering cells. Ultrastructural immunocytochemistry indicated surface binding of transferrin by cells in the gels, and the presence of endogenous transferrin on the surfaces of mesoderm cells in situ and in their extracellular environment. It is suggested that by binding to cell surface receptors, transferrin may be able to influence the strength of cellular adhesion to collagen and hence the capacity for cell locomotion.

scholarly journals The Sp1-Responsive microRNA-15b Negatively Regulates Rhabdovirus-Triggered Innate Immune Responses in Lower Vertebrates by Targeting TBK1

2021 ◽  
Vol 11 ◽  
Author(s):  
Renjie Chang ◽  
Qing Chu ◽  
Weiwei Zheng ◽  
Lei Zhang ◽  
Tianjun Xu
Keyword(s):  
Immune Response ◽  
Immune Responses ◽  
Innate Immune ◽  
Infection Model ◽  
Regulation Mechanism ◽  
Type I ◽  
Innate Immune Responses ◽  
Positive Role ◽  
Siniperca Chuatsi ◽  
Antiviral Immune Response

As is known to all, the production of type I interferon (IFN) plays pivotal roles in host innate antiviral immunity, and its moderate production play a positive role in promoting the activation of host innate antiviral immune response. However, the virus will establish a persistent infection model by interfering with the production of IFN, thereby evading the organism inherent antiviral immune response. Therefore, it is of great necessity to research the underlying regulatory mechanisms of type I IFN appropriate production under viral invasion. In this study, we report that a Sp1–responsive miR-15b plays a negative role in siniperca chuatsi rhabdovirus (SCRV)-triggered antiviral response in teleost fish. We found that SCRV could dramatically upregulate miiuy croaker miR-15b expression. Enhanced miR-15b could negatively regulate SCRV-triggered antiviral genes and inflammatory cytokines production by targeting TANK-binding kinase 1 (TBK1), thereby accelerating viral replication. Importantly, we found that miR-15b feedback regulates antiviral innate immune response through NF-κB and IRF3 signaling pathways. These findings highlight that miR-15b plays a crucial role in regulating virus–host interactions, which outlines a new regulation mechanism of fish’s innate immune responses.

Feeding experiments of the seep-associated foraminifer Nonionellina labradorica with a marine methanotroph from the Arctic

2021 ◽  
Author(s):  
Christiane Schmidt ◽  
Geslin Emmanuelle ◽  
Bernhard Joan M. ◽  
LeKieffre Charlotte ◽  
Roberge Helene ◽  
...  
Keyword(s):  
Barents Sea ◽  
Feeding Habits ◽  
The Arctic ◽  
Type I ◽  
Intracytoplasmic Membrane ◽  
Organic Detritus ◽  
Feeding Experiments ◽  
Food Vacuoles ◽  
Species Specific

<p>Foraminifera on the seafloor are known to have species-specific feeding habits. Among those are deposit feeders, eating organic detritus and bacteria. Little is known about the feeding habits of foraminifera from Arctic seep environments. That is, in particular, of interest as variable δ<sup>13</sup>C values in the tests of foraminifera have been suggested to be partly linked with a diet rich in bacteria, themselves lighter in δ<sup>13</sup>C values. As there is little information on the ecology of the foraminifer <em>Nonionellina labradorica</em> (Dawson, 1860), this study examined feeding habits on bacteria and compared them to in situ collected specimens, using Transmission Electron microscopy (TEM). As bacterial food, the marine methane-oxidizing bacterium <em>Methyloprofundus sedimenti</em> was chosen, which is an important representative of methanotrophs in the marine environment near methane seeps. Sediment samples containing living N. labradorica specimens collected in close vicinity(approx. 5 m) from an active methane seep in Storfjordrenna, Barents Sea (382-m water depth).  We performed a feeding experiment on <em>N. labradorica </em>(n=17 specimen), which were incubated in the dark at in situ temperature. Specimens were fed at the beginning of the experiment, except the un-fed controls, and incubations terminated after 4, 8 and 20 h. After fixation in epoxy resin the ultrastructure of all specimens and their food vacuoles was observed and compared using a TEM. All examined specimens were living at the time of fixation, based on observation of intact mitochondrial membranes. In all specimens, inorganic detritus was preserved inside food vacuoles. Closer observation of food vacuoles also revealed that in addition to inorganic debris, such as clay, occasionally bacteria were visible. This led us to conclude that our <em>N. labradorica </em>can  generally be classified as a deposit feeder, which is rather a generalist than a specialist. Regarding uptake of <em>M. sedimenti</em>, the timing of the experimentation seemed to be critical. We did not observe methanotrophs preserved in the resin at the 4 and 8 h incubations, but found two putative methanotrophs near the apertural region after the 20-h incubation. After closer observation, we could identify one of those two putative specimen as the menthanothroph <em>M. sedimenti</em> near the foraminiferal aperture, based on presence of a typical type I stacked intracytoplasmic membrane (ICM) and storage granules (SC). We concluded that <em>N. labradorica</em> may ingest <em>M. sedimenti</em> via “untargeted grazing” in seeps. Further studies must examine the exact relationship between diet and δ<sup>13</sup>C in foraminiferal test on several different paleo-oceanographically relevant species.</p>

Establishment of Full-length cDNA Clones and an Efficient Oral Infection Model for Feline Coronavirus in Cats

2021 ◽  
Author(s):  
Gang Wang ◽  
Guangli Hu ◽  
Rui Liang ◽  
Jiale Shi ◽  
Xiuxiu Qiu ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Full Length ◽  
Infection Model ◽  
Type I ◽  
Feline Infectious Peritonitis ◽  
Spike Gene ◽  
Full Length Cdna ◽  
Adult Cats

Feline infectious peritonitis virus (FIPV) is the etiologic agent of feline infectious peritonitis (FIP) and causes fatal disease in cats of almost all ages. Currently, there are no clinically approved drugs or effective vaccines for FIP. Furthermore, the pathogenesis of FIP is still not fully understood. There is an urgent need for an effective infection model of feline infectious peritonitis induced by FIPV. Here, we constructed a field type I FIPV full-length cDNA clone, pBAC-QS, corresponding to the isolated FIPV QS. By replacing the FIPV QS spike gene with the commercially available type II FIPV 79-1146 (79-1146_CA) spike gene, we established and rescued a recombinant virus, designated rQS-79. Moreover, we constructed 79-1146_CA infectious full-length cDNA pBAC-79-1146_CA, corresponding to recombinant FCoV 79-1146_CA (r79-1146_CA). In animal experiments with one- to two-year-old adult cats orally infected with the recombinant virus, rQS-79 induced typical FIP signs and 100% mortality. In contrast to cats infected with rQS-79, cats infected with 79-1146_CA did not show obvious signs. Furthermore, by rechallenging rQS-79 in surviving cats previously infected with 79-1146_CA, we found that there was no protection against rQS-79 with different titers of neutralizing antibodies. However, high titers of neutralizing antibodies may help prolong the cat survival time. Overall, we report the first reverse genetics of virulent recombinant FCoV (causing 100% mortality in adult cats) and attenuated FCoV (causing no mortality in adult cats), which will be powerful tools to study the pathogenesis, antiviral drugs and vaccines for FCoV. Importance Tissue- or cell culture-adapted feline infectious peritonitis virus (FIPV) usually loses pathogenicity. To develop a highly virulent FIPV, we constructed a field isolate type I FIPV full-length clone with the spike gene replaced by the 79-1146 spike gene, corresponding to a virus named rQS-79, which induces high mortality in adult cats. rQS-79 represents the first described reverse genetics system for highly pathogenic FCoV. By further constructing the cell culture-adapted FCoV 79-1146_CA, we obtained infectious clones of virulent and attenuated FCoV. By in vitro and in vivo experiments, we established a model that can serve to study the pathogenic mechanisms of FIPV. Importantly, the wild-type FIPV replicase skeleton of serotype I will greatly facilitate the screening of antiviral drugs, both in vivo and in vitro.

Exercise-induced cellular alterations in the diaphragm

1992 ◽  
Vol 263 (5) ◽  
pp. R1093-R1098 ◽  
Author(s):  
S. K. Powers ◽  
D. Criswell ◽  
F. K. Lieu ◽  
S. Dodd ◽  
H. Silverman
Keyword(s):  
Exercise Training ◽  
Endurance Training ◽  
Fiber Type ◽  
Capillary Density ◽  
Oxidative Capacity ◽  
Control Group ◽  
Type I ◽  
Fiber Types ◽  
Image Analysis System ◽  
Cross Sectional

Limited data exist concerning the effects of exercise training on cellular oxidative capacity in the diaphragm of senescent animals. In this study we examined the changes in cellular oxidative capacity, muscle cell cross-sectional area (CSA), and capillarity within the costal diaphragm of senescent animals after a 10-wk endurance-training program. Twelve 24-mo-old female Fischer 344 rats were divided into either a sedentary control group (n = 6) or exercise training group (n = 6). The trained animals exercised on a motor-driven treadmill (60 min/day, 5 days/wk) at a work rate equal to approximately 55-65% VO2max. Capillaries were identified histologically and fiber types determined using adenosinetriphosphatase (ATPase) histochemistry. Succinate dehydrogenase (SDH) activity and CSA in individual fibers were measured using a computerized image analysis system. Exercise training did not increase (P > 0.05) the capillary-to-fiber ratio for any fiber type. However, training significantly decreased CSA (P < 0.05) and increased capillary density (capillary number/CSA) (P < 0.05) in type I, type IIa, and type IIb fibers. Furthermore, exercise training resulted in small but significant increase in SDH activity (P < 0.05) in type I and IIa fibers, whereas training did not alter SDH activity (P > 0.05) in type IIb fibers. These data demonstrate that endurance training in senescent animals results in small relative improvements in both oxidative capacity and capillary density in costal diaphragmatic type I and IIa muscle fibers. The increase in both capillary density and fiber SDH activity was largely due to a reduction in fiber CSA.

scholarly journals Human coronaviruses 229E and OC43 replicate and induce distinct antiviral responses in differentiated primary human bronchial epithelial cells

2020 ◽  
Vol 319 (6) ◽  
pp. L926-L931
Author(s):  
Su-Ling Loo ◽  
Peter A. B. Wark ◽  
Camille Esneau ◽  
Kristy S. Nichol ◽  
Alan C-Y. Hsu ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Time Course ◽  
Bronchial Epithelial Cells ◽  
Innate Immune ◽  
Infection Model ◽  
Type I ◽  
Human Bronchial Epithelial Cells ◽  
Cell Infection ◽  
Bronchial Epithelial ◽  
Human Coronaviruses

The recurrent emergence of novel, pathogenic coronaviruses (CoVs) severe acute respiratory syndrome coronavirus 1 (SARS-CoV-1; 2002), Middle East respiratory syndrome (MERS)-CoV (2012), and most recently SARS-CoV-2 (2019) has highlighted the need for physiologically informative airway epithelial cell infection models for studying immunity to CoVs and development of antiviral therapies. To address this, we developed an in vitro infection model for two human coronaviruses; alphacoronavirus 229E-CoV (229E) and betacoronavirus OC43-CoV (OC43) in differentiated primary human bronchial epithelial cells (pBECs). Primary BECs from healthy subjects were grown at air-liquid interface (ALI) and infected with 229E or OC43, and replication kinetics and time-course expression of innate immune mediators were assessed. OC43 and 229E-CoVs replicated in differentiated pBECs but displayed distinct replication kinetics: 229E replicated rapidly with viral load peaking at 24 h postinfection, while OC43 replication was slower peaking at 96 h after infection. This was associated with diverse antiviral response profiles defined by increased expression of type I/III interferons and interferon-stimulated genes (ISGs) by 229E compared with no innate immune activation with OC43 infection. Understanding the host-virus interaction for previously established coronaviruses will give insight into pathogenic mechanisms underpinning SARS-CoV-2-induced respiratory disease and other future coronaviruses that may arise from zoonotic sources.

Rat diaphragm during postnatal development. I. Changes in distribution of muscle fibre type and in oxidative potential

1996 ◽  
Vol 8 (3) ◽  
pp. 391 ◽  
Author(s):  
MD Fratacci ◽  
M Levame ◽  
A Rauss ◽  
H Bousbaa ◽  
G Atlan
Keyword(s):  
Muscle Fibre ◽  
Fibre Type ◽  
Oxidative Capacity ◽  
Cross Sectional Area ◽  
Fibre Types ◽  
Type I ◽  
Sectional Area ◽  
Rat Diaphragm ◽  
Oxidative Potential ◽  
Cross Sectional

The changes occurring in the histochemical characteristics of the rat diaphragm during the postnatal period were examined. Fibre-type distribution, fibre oxidative capacity, i.e. succinate-dehydrogenase (SDH) activity, and cross-sectional area were compared in the costal (COS) and crural (CRU) regions, and across their abdominal and thoracic surfaces. The proportions of type I and IIb fibres in both COS and CRU increased with age, while the proportion of type IIa fibres progressively decreased. For COS, fibre distribution was homogeneous over the entire muscle and did not change after 4 weeks. For CRU, it was heterogeneous with a higher proportion of type I fibres on the thoracic surface as from the first week. All fibre types significantly increased in cross-sectional area between 1 and 8 weeks, with no significant differences in COS and CRU. Mean SDH activity did not differ between COS and CRU or across the muscles. Mean SDH activities-were low and identical in all fibre types at birth, and then increased, peaking at the 6th week in type I and IIa fibres. When total muscle fibre oxidative capacity was calculated from an index including fibre-type proportion, cross-sectional area and mean SDH activity, it was significantly higher at 1 than at 8 weeks after birth; this might have functional implications for the newborn.

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