Silk fibroin (SF) microparticles were administered in the diet of gilthead seabream with or without experimental skin wounds to study the effects on cellular immunity and liver. A commercially ...available diet was enriched with varying amount of SF: 0, 50 and 100 mg kg−1 (representing the control, SF50 and SF100 diets respectively). The animals were fed for 30 days and half of them were sampled. Similar experimental wounds were then performed on the rest of fish, and they continued to be fed the same diet. At 7 days post‐wounding, samples were taken from the wounded fish. Cellular immunity was studied on head kidney leucocytes (phagocytosis, respiratory and peroxidase content) and liver status (histological study and gene expression) were studied. Our results showed that experimental wounds affect both cellular immunity (by decreasing leucocyte respiratory burst and peroxidase activity) and altered liver histology (by inducing vascularisation and congestion of blood vessels). In addition, it influences the expression of genes that serve as markers of oxidative stress, endoplasmic reticulum stress and apoptosis. The highest dose of SF (SF100) increased the phagocytic capacity of leucocytes the most, as well as the expression of genes related to blood vessel formation in the liver. Furthermore, increased expression of antioxidant genes (cat and gsr) and decreased expression of genes related to reticulum endoplasmic stress (grp94 and grp170) and apoptosis (nos and jnk) were detected in these fish fed with SF100 and wounded. In conclusion, fed fish with SF100 had many beneficial effects as cellular immunostimulant and hepatoprotection in wounded fish. Its use could be of great interest for stress management in farmed fish conditions.
The effect of the dietary incorporation of drumstick,
Moringa oleifera
, leaf meal (MOL; 0, 5, 10 and 15%) on the growth, feed utilization, some skin mucus and systemic immune parameters and ...intestinal immune-related gene expression in gilthead seabream (
Sparus aurata
) specimens. The experiment lasted 4 weeks. The results revealed that MOL can be incorporated in
S. aurata
diet up to 10% with no significant negative effect on growth and feed utilization. However, there was a significant decrease with MOL at a level of 15% after 2 weeks of feeding. The systemic immune status of fish fed with the different levels of MOL showed an improvement in head kidney leucocyte phagocytosis, respiratory burst and peroxidase activities. Also, serum humoral components, including protease, ACH
50
and lysozyme activities and IgM level, increased with MOL inclusion especially at the 5% level. MOL at 5% improved skin-mucosal immunity such as protease, antiprotease, peroxidase and lysozyme activities. Moreover, the feeding of MOL revealed an upregulation of the intestinal mucosal immunity genes (
lyso
and
c3
), tight junction proteins (
occludin
and
zo-1
) and anti-inflammatory cytokines (
tgf-β
) with a downregulation of pro-inflammatory cytokine (
tnf-α
). Therefore, it is recommended to incorporate MOL in
S. aurata
diets at a level of 5% for the best immune status or 10% for the high growth performance and acceptable immune surveillance.
Graphical abstract
ᅟ
In aquatic animals, the mucosal barrier is the first line of innate immune defence against external chemicals and pathogens. In this study, the effects of dietary
Moringa oleifera
leaf (MOL) ...supplementation on skin and gill mucosal immunity, antioxidants and stress responses were evaluated in seabream (
Sparus aurata
) fingerlings exposed to hydrogen peroxide (H
2
O
2
)
. A total of 144 specimens (10.11 ± 0.41 g) were divided into four treatments (three replicates per treatment contained 12 specimens each) and fed a non-supplemented control diet or a 1, 2.5 or 5% MOL-supplemented diet. After three weeks of feeding, six specimens from each aquarium were sampled for blood, mucus and tissues. The other six fish in each aquarium were subjected to H
2
O
2
exposure. The results revealed that MOL did not negatively affect either cortisol or glucose levels. MOL supplementation significantly (
P
< 0.05) improved skin mucosal immunity-related characteristics, including phosphatase, peroxidase and lysozyme activity and IgM levels. Additionally, MOL upregulated the expression of antioxidant genes (
sod
and
cat
), an anti-inflammatory gene (
tgf-β
), tight junction protein genes (
occludin
and
zo-1
),
c3,
and
igm
in both the skin and gills. However, H
2
O
2
exposure significantly (
P
< 0.05) increased both cortisol and glucose levels and disrupted skin mucosal immune function by significantly (
P
< 0.05) decreasing phosphatase, peroxidase, protease, antiprotease and lysozyme activity and IgM levels. H
2
O
2
exposure severely decreased the mRNA levels of the studied genes. MOL dietary supplementation at the 5% level successfully attenuated the negative effects of H
2
O
2
on the mucosal immune response in both the skin and gills. In conclusion, dietary MOL supplementation at the 5% level is recommended to improve
S. aurata
mucosal immune function under both normal and stress conditions. Additionally, exposure to H
2
O
2
disrupts the mucosal immunity of fish. This contributes knowledge on the routes involved in mucosal innate immunity and could help to understand the fish resistance against chemicals exposure.
Graphical abstract
The impact of dietary supplementation with silk fibroin (SF) microparticles on the wound healing process in gilthead seabream (Sparus aurata) skin was studied. A control diet was enriched with ...different SF levels: 0 (control), 50 (SF50 diet), and 100 (SF100 diet) mg Kg−1 to form three experimental diets and was fed to seabream for 30 days. Experimental wounds were performed and after 7 days post-wounding (dpw) skin mucus immunity, macroscopic wound closure, and skin regeneration were studied at a microscopic and genetic level. Results indicated that fish fed SF100 did not suffer the decreases in protease and IgM levels observed in the skin mucus of wounded fish fed with the control diet. Macroscopic findings illustrated that dietary SF100 significantly improved the wound closure ratio compared to those reared in the control group. At a microscopic level, changes in the shape of keratocyte cells were evident in the wounded fish. In addition, the intercellular spaces present between epidermal cells and their proliferation in the epidermis, as well as the presence of blood vessels in the dermis were significantly statistically higher in the skin of fish fed the SF100 diet and sampled at 7 dpw compared to those observed in the skin of fish fed the control or SF50 diets. Moreover, regarding the RNA: DNA ratio, statistically significant increases and decreases were observed in fish fed the control and SF100 diet, respectively, in non-wounded and wounded fish. Interestingly, dietary SF100 supplementation improved skin cell proliferation, enhanced the inflammatory phase, and increased the expression of important genes involved in tissue repair and extracellular matrix formation. In conclusion, the SF100 diet can be considered as an appropriate feed additive to improve wound healing in gilthead seabream.
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•The impacts of dietary supplementation with silk fibroin microparticles (SF) on the skin wound healing process were studied.•The macroscopic wound closure and the skin regeneration at a microscopic and genetic level were studied.•At microscopic levels, changes in the keratocyte cells shape were evident in wounded fish.•Dietary SF100 supplementation improved the skin cell proliferation and enhanced the inflammatory phase.
Okra, Abelmoschus esculentus L., is a popular vegetable crop with many bioactive compounds. The in vitro immunostimulant, cytotoxic, bactericidal and antioxidant activities of ethanolic extracts ...obtained from different parts of okra (leaves, fruits, and seeds) were studied. Phytochemical screening of hydroalcoholic extracts of okra leaves, fruits and seeds revealed a significant content of total phenols and flavonoids. Notable effects on the activities of leukocytes in the head kidney of European sea bass (Dicentrarchus labrax) (viability, phagocytic ability and capacity, and respiratory burst), as well as on peroxidase leukocyte contents were detected after incubation for 24 h with different concentrations (0.01–1 mg mL−1) of the extracts. The mean concentrations (0.1 and 0.5 mg mL−1) of the different extracts increased the phagocytic ability and respiratory activity of head kidney leukocytes. However, the mean concentrations (0.1 mg mL−1) of leaf and fruit extracts significantly decreased the peroxidase activity of leukocytes. In addition, all ethanolic okra extracts at higher concentrations (1 mg mL−1) produced a marked reduction in the viability of the DLB-1 cell line compared to the viability recorded in the control samples. In addition, ethanolic extracts used at 0.5 and 1 mg mL−1 had a significant cytotoxic effect on the viability of PLHC-1 cells. Finally, all doses of seed and leaf extracts at higher concentrations (0.5 and 1 mg mL−1) showed significant bactericidal activity on two fish pathogenic bacteria, Vibrio anguillarum and V. harveyi strains. Finally, a remarkable antioxidant activity was detected on the ethanolic extracts. All these results point to their possible use as an alternative to chemical compounds in farmed fish.
Vibriosis is an infectious disease that generates large economic losses in Mediterranean aquaculture. Vibrio harveyi is one of the marine bacteria causing this disease, it is widespread in the ...Mediterranean Sea and causes ulcers on the skin of the fish it infects. In addition, the skin is a route of entry and colonization of this pathogen. In this study, one group of fish was injected intraperitoneally with phosphate buffered saline (control group) and another with V. harveyi (infected group). At 4 h after injection, samples of skin mucus, blood, skin, head kidney, liver, and spleen were collected to study the immune response generated. Liver histology showed notable alterations in hepatocyte morphology, such as increased vacuolization. Bactericidal activity was measured in skin mucus and serum against V. harveyi and V. anguillarum, different changes in this activity were recorded depending on the bacteria target and sample (skin mucus or serum) used. Gene expression of genes encoding hepcidins and piscidins (antimicrobial peptides) was performed in the mentioned organs. The results indicated a different expression according to the type of AMP and the tissue studied. Hepcidin appeared involved in all tissues studied while piscidins were in the spleen. In this study we have integrated hepcidin-piscidin modulation with the effects of infection on skin mucosa, serum and hepatocyte morphology. Knowing the changes produced in all these parameters improves the understanding of the infection in the first hours in sea bream and could have applications in the diagnosis or treatment of vibriosis in fish farms.
•Vibrio harveyi infection has been achieved and studied in gilthead seabream (Sparus aurata).•Increase vacuolization of hepatocytes was evident in infected seabream at 4 h post-injection.•Antibacterial activity of skin mucus and serum was decreased in infected fish.•Differential gene expression of AMPs genes reinforce the role of skin, spleen and head kidney in bacterial infection.•Expression patterns of hepcidins and piscidins were different in infected animals.
Objective: Systemic lupus erythematosus (SLE) is associated to boosted atherosclerosis development and a higher cardiovascular disease risk. This study aimed to delineate the role of anti-double ...stranded DNA (anti-dsDNA) antibodies on the molecular profile and the activity of immune and vascular cells, as well as on their enhanced cardiovascular risk. Approach and Results: Eighty SLE patients were included. Extensive clinical/analytical evaluation was performed, including cardiovascular disease parameters (endothelial function, proatherogenic dyslipidemia, and carotid intima-media thickness). Gene and protein expression profiles were evaluated in monocytes from patients diagnosed positive or negative for anti-dsDNA antibodies by using NanoString and cytokine arrays, respectively. NETosis and circulating inflammatory profile was assessed in both neutrophils and plasma. Positivity and persistence of anti-dsDNA antibodies in SLE patients were associated to endothelial dysfunction, proatherogenic dyslipidemia, and accelerated atherosclerosis. In parallel, anti-dsDNA antibodies were linked to the aberrant activation of innate immune cells, so that anti-dsDNA(+) SLE monocytes showed distinctive gene and protein expression/activity profiles, and neutrophils were more prone to suffer NETosis in comparison with anti-dsDNA(−) patients. Anti-dsDNA(+) patients further displayed altered levels of numerous circulating mediators related to inflammation, NETosis, and cardiovascular risk. In vitro, Ig-dsDNA promoted NETosis on neutrophils, apoptosis on monocytes, modulated the expression of inflammation and thrombosis-related molecules, and induced endothelial activation, at least partially, by FcR (Fc receptor)-binding mechanisms. Conclusions: Anti-dsDNA antibodies increase the cardiovascular risk of SLE patients by altering key molecular processes that drive a distinctive and coordinated immune and vascular activation, representing a potential tool in the management of this comorbidity.
Abstract
Aims
The interplay between aortic stenosis (AS), cardiovascular events, and mortality is poorly understood. In addition, how echocardiographic indices compare for predicting outcomes remains ...unexplored for the full range of AS severity.
Methods and results
We prospectively calculated peak jet velocity (Vmax) and aortic valve area (AVA) in 5994 adult subjects with and without AS. We linked ultrasound data to 5-year mortality and clinical events obtained from electronic medical records. Proportional-hazard and negative binomial regression models were adjusted for relevant covariables such as age, sex, comorbidities, stroke-volume, LV ejection fraction, left valve regurgitation, aortic valve sclerosis or calcification, and valve replacement. We observed a strong linear relationship between Vmax and all-cause mortality (hazard ratio: 1.26, 95% confidence interval: 1.19–1.33 per 100 cm/s), cardiovascular events, as well as incidental and recurrent heart failure (HF). Adjusted risks were highly significant even at Vmax values in the range of 150–200 cm/s, risk curves separating very early after the index exam. Vmax was not associated with coronary, arrhythmic, cerebrovascular, or non-cardiovascular events. Although risks were confirmed when AVA was entered in place of Vmax, the risks estimated for categories based on the two indices were mismatched, even in patients with normal flow. An external cohort comprising 112 690 patients confirmed augmented risks of all-cause and cardiovascular mortality starting at values of Vmax and AVA in the range of mild AS.
Conclusions
Aortic stenosis is strongly associated to all-cause mortality, cardiovascular mortality, and cardiac events, specifically HF. Risks increase in parallel to the degree of outflow obstruction but are apparent very early in patients with mild disease. Criteria for grading AS based on Vmax and AVA are mismatched in terms of outcomes.
Graphical Abstract
Mitochondria are a crucial cellular organelle that organizes a wide range of biological processes, from energy production and calcium homeostasis to cell proliferation, differentiation, and death as ...well as inflammation. Mitochondria also support immune cell phenotypes and function. The aim of this communication is to evaluate the mitochondrial status and plasticity of four fish cell lines: SAF-1 (derived from gilthead sea bream skin), DLB-1 (derived from European sea bass brain), FuB-1 (established from mummichog brain), and PLHC-1 (a topminnow hepatocellular carcinoma). A mitochondrial stress assay was performed to compare the bioenergetic capabilities of these four fish cell lines as well as the importance of choosing the correct cell line when performing different in vitro studies.
Patients with nonischemic dilated cardiomyopathy (NIDCM) are at higher risk of stroke than their age-matched population. However, the risk of bleeding neutralizes the benefit of preventive oral ...anticoagulation. In this work, we show that in patients in sinus rhythm, the burden of stroke is related to intraventricular stasis metrics derived from echocardiography. Therefore, stasis metrics may be useful to personalize primary prevention anticoagulation in these patients.
Cardioembolic stroke is one of the most devastating complications of nonischemic dilated cardiomyopathy (NIDCM). However, in clinical trials of primary prevention, the benefits of anticoagulation are hampered by the risk of bleeding. Indices of cardiac blood stasis may account for the risk of stroke and be useful to individualize primary prevention treatments. We performed a cross-sectional study in patients with NIDCM and no history of atrial fibrillation (AF) from two sources: 1) a prospective enrollment of unselected patients with left ventricular (LV) ejection fraction <45% and 2) a retrospective identification of patients with a history of previous cardioembolic neurological event. The primary end point integrated a history of ischemic stroke or the presence intraventricular thrombus, or a silent brain infarction (SBI) by imaging. From echocardiography, we calculated blood flow inside the LV, its residence time ( T R ) maps, and its derived stasis indices. Of the 89 recruited patients, 18 showed a positive end point, 9 had a history of stroke or transient ischemic attack (TIA) and 9 were diagnosed with SBIs in the brain imaging. Averaged T R , Formula: see text performed well to identify the primary end point AUC (95% CI) = 0.75 (0.61–0.89), P = 0.001. When accounting only for identifying a history of stroke or TIA, AUC for Formula: see text was 0.92 (0.85–1.00) with odds ratio = 7.2 (2.3–22.3) per cycle, P < 0.001. These results suggest that in patients with NIDCM in sinus rhythm, stasis imaging derived from echocardiography may account for the burden of stroke. NEW & NOTEWORTHY Patients with nonischemic dilated cardiomyopathy (NIDCM) are at higher risk of stroke than their age-matched population. However, the risk of bleeding neutralizes the benefit of preventive oral anticoagulation. In this work, we show that in patients in sinus rhythm, the burden of stroke is related to intraventricular stasis metrics derived from echocardiography. Therefore, stasis metrics may be useful to personalize primary prevention anticoagulation in these patients.
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