Up-regulation of inflammatory, oxidative stress, and apoptotic mediators via inflammatory, oxidative stress, and apoptosis-associated pathways in bovine endometritis.

Endometritis is the inflammation of the endothelial lining of the uterine lumen and is multifactorial in etiology. Escherichia (E.) coli is a Gram-negative bacteria, generally considered as a primary causative agent for bovine endometritis. Bovine endometritis is characterized by the activation of Toll-like receptors (TLRs) by E. coli, which in turn triggers inflammation, oxidative stress, and apoptosis. The objective of this study was to investigate the gene expression of inflammatory, oxidative stress, and apoptotic markers related to endometritis in the uteri of cows. Twenty uterine tissues were collected from the abattoir. Histologically, congestion, edema, hyperemia, and hemorrhagic lesions with massive infiltration of neutrophil and cell necrosis were detected markedly (P < 0.05) in infected uterine samples. Additionally, we identify E. coli using the ybbW gene (177 base pairs; E. coli-specific gene) from infected uterine samples. Moreover, qPCR and western blot results indicated that TLR2, TLR4, proinflammatory mediators, and apoptosis-mediated genes upregulated except Bcl-2, which is antiapoptotic, and there were downregulations of oxidative stress-related genes in the infected uterine tissue. The results of our study suggested that different gene expression regimes related to the immune system reflex were activated in infected uteri. This research gives a novel understanding of active immunological response in bovine endometritis.

Icariin Alleviates Escherichia coli Lipopolysaccharide-Mediated Endometritis in Mice by Inhibiting Inflammation and Oxidative Stress.

Icariin (ICA) is a naturally occurring phytochemical agent primarily extracted from Epimedium Brevicornum Maxim (Family Berberidaceae) with a broad spectrum of bioactivities. Endometritis is a uterine disease that causes enormous losses in the dairy industry worldwide. In this study, anti-inflammatory and anti-oxidant properties of ICA were investigated against lipopolysaccharide (LPS)-induced endometritis in mice to investigate possible underlying molecular mechanisms. Sixty heathy female Kunming mice were randomly assigned to four groups (n = 15), namely control, LPS, LPS + ICA, and ICA groups. The endometritis was induced by intrauterine infusion of 50 µL of LPS (1 mg/mL). After 24 h of onset of LPS-induced endometritis, ICA groups were injected thrice by ICA intraperitoneally six hours apart. Histopathological examination, enzyme linked immunosorbent assay (ELISA), real time quantitative polymerase chain reaction (RT-qPCR), western blotting, and immunohistochemistry were used in this study. Histological alterations revealed that ICA markedly mitigated uterine tissue injury caused by LPS. The results showed that the ICA inhibited the production of pro-inflammatory cytokines (IL-1ß, IL-6, and TNF-α) and boosted the production of anti-inflammatory cytokines (IL-10). Additionally, ICA modulated the expression of malondialdehyde (MDA), reactive oxygen species (ROS), superoxide dismutase 1 (SOD1), catalase (CAT), and glutathione peroxidase 1 (Gpx1) induced by LPS. The administration of ICA significantly (p < 0.05) improved the mRNA and protein expression of Toll-like receptor (TLR) 4. The western blotting and ELISA finding revealed that the ICA repressed LPS-triggered NF-κB pathway activation. Moreover, ICA improved the antioxidant defense system via activation of the Nrf2 pathway. The results revealed that ICA up-regulated the mRNA and protein expression of Nuclear erythroid-2-related factor (Nrf2), NAD(P)H: quinone oxidoreductase 1 (NQO1), heme oxygenase-1 (HO-1), and glutamate-cysteine ligase catalytic subunit (GCLC) under LPS exposure. Conclusively, our findings strongly suggested that ICA protects endometritis caused by LPS by suppressing TLR4-associated NF-κB and Nrf2 pathways. Altogether, these innovative findings may pave the way for future studies into the therapeutic application of ICA to protect humans and animals against endometritis.

Upregulated-gene expression of pro-inflammatory cytokines, oxidative stress and apoptotic markers through inflammatory, oxidative and apoptosis mediated signaling pathways in Bovine Pneumonia.

Pneumonia is the acute inflammation of lung tissue and is multi-factorial in etiology. Staphylococcus aureus (S. aureus) is a harmful pathogen present as a normal flora of skin and nares of dairy cattle. In bovine pneumonia, S. aureus triggers to activates Toll-Like Receptors (TLRs), that further elicits the activation of the inflammation via NF-κB pathway, oxidative stress and apoptotic pathways. In the current study, pathogen-associated gene expression of the pro-inflammatory cytokines, oxidative stress and apoptotic markers in the lung tissue of cattle was explored in bovine pneumonia. Fifty lung samples collected from abattoir located in Wuhan city, Hubei, China. Histopathologically, thickening of alveolar wall, accumulation of inflammatory cells and neutrophils in perivascular space, hyperemia, hemorrhages and edema were observed in infected lungs as compared to non-infected lung samples. Furthermore, molecular identification and characterization were carried by amplification of S. aureus-specific nuc gene (270 base pairs) from the infected and non-infected lung samples to identify the S. aureus. Moreover, qPCR results displayed that relative mRNA levels of TLR2, TLR4, pro-inflammatory gene (IL-1ß, IL-6 and TNF-α) and apoptosis-associated genes (Bax, caspase-3 and caspase-9) were up-regulated except Bcl-2, which is antiapoptotic in nature, and oxidative stress related genes (Nrf2, NQO1, HO-1 and GCLC) which was down-regulated in infected pulmonary group. The relative protein expression of NF-κB, mitochondria-mediated apoptosis gene was up-regulated while Bcl-2 and Nrf2 pathway genes were downregulated in infected cattle lungs. Our findings revealed that genes expression levels of inflammatory mediators, oxidative stress and apoptosis were associated with host immunogenic regulatory mechanisms in the lung tissue during infection. Conclusively, the present study provides insights of active immune response via TLRs-mediated inflammatory, oxidative damage, and apoptotic paradox.

Ginsenoside Rb1 protects from Staphylococcus aureus-induced oxidative damage and apoptosis through endoplasmic reticulum-stress and death receptor-mediated pathways.

Acute lung injury (ALI) is acute uncontrolled inflammation of lung tissue that leads to high fatality both in human and animals. Staphylococcus aureus (S. aureus) could be an opportunistic, versatile bacterial etiology of ALI. Ginsenoside Rb1 (Rb1) is extracted from the Panax ginseng, which displays a wide range of biological and pharmacological effects. However, protective effects of Rb1 in S. aureus-induced ALI though endoplasmic reticulum (ER) stress and death receptor-mediated pathways have not yet been reported. Therefore, present study was planned with the aims to investigate the antioxidant and anti-apoptotic properties of Rb1 through regulation of ER stress as well as death receptor-mediated pathways in ALI induced by S. aureus in mice. In this study, four groups of healthy Kunming mice (n = 48) were used. The S. aureus (80 µl; 1 ×107 CFU/10 µl) was administered intranasally to establish mice model of ALI. After 24 h of onset of S. aureus-induced ALI, the mice were injected thrice with Rb1 (40 mg/kg) intraperitoneally six hours apart. Histopathology, enzyme linked immunosorbent assay (ELISA), real time quantitative polymerase chain reaction (RT-qPCR), Immunohistochemistry and western blotting assay were employed in the current study. Our results suggested that Rb1 administration save lungs from pulmonary injury by reducing wet to dry (W/D) ratio, protein levels, total cells, neutrophilic count, reactive oxygen species (ROS), myeloperoxidase (MPO), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (Gpx)1 depletion. Meanwhile, Rb1 therapy ameliorated histopathology alteration of lung tissue and pro-inflammatory cytokines secretion. The gene expression of ER stress marker (PERK, AFT-6, IRE1 and CHOP) were upregulated markedly (P < .05) in S. aureus-instilled groups, which was reduced by Rb1 administration that is reveled from the result findings of the RT-qPCR and immunoblot assay. The results of immunohistochemistry for CHOP indicated the increased expression in S. aureus groups which in turn ameliorated by Rb1 treatment. The mRNA expression demonstrated that death receptor-associated genes (FasL, Fas, FADD and caspase-8) showed up-regulation in S. aureus group. The similar findings were observed for the protein expression of caspase-8, FADD and Fas. Rb1 treatment markedly (P < .05) reversed protein and mRNA expression levels of these death receptor-associated genes when compared to the S. aureus group. Taken together, Rb1 attenuated S. aureus-induced oxidative damage via the ER stress-mediated pathway and apoptosis through death receptor-mediated pathway. Conclusively, our findings provide an insight into preventive mechanism of Rb1 in ALI caused by S. aureus and hence proven a scientific baseline for the therapeutic application of Rb1.

Ginsenoside Rb1 Mitigates Escherichia coli Lipopolysaccharide-Induced Endometritis through TLR4-Mediated NF-κB Pathway.

Endometritis is the inflammatory response of the endometrial lining of the uterus and is associated with low conception rates, early embryonic mortality, and prolonged inter-calving intervals, and thus poses huge economic losses to the dairy industry worldwide. Ginsenoside Rb1 (GnRb1) is a natural compound obtained from the roots of Panax ginseng, having several pharmacological and biological properties. However, the anti-inflammatory properties of GnRb1 in lipopolysaccharide (LPS)-challenged endometritis through the TLR4-mediated NF-κB signaling pathway has not yet been researched. This study was planned to evaluate the mechanisms of how GnRb1 rescues LPS-induced endometritis. In the present research, histopathological findings revealed that GnRb1 ameliorated LPS-triggered uterine injury. The ELISA and RT-qPCR assay findings indicated that GnRb1 suppressed the expression level of pro-inflammatory molecules (TNF-α, IL-1ß and IL-6) and boosted the level of anti-inflammatory (IL-10) cytokine. Furthermore, the molecular study suggested that GnRb1 attenuated TLR4-mediated NF-κB signaling. The results demonstrated the therapeutic efficacy of GnRb1 in the mouse model of LPS-triggered endometritis via the inhibition of the TLR4-associated NF-κB pathway. Taken together, this study provides a baseline for the protective effect of GnRb1 to treat endometritis in both humans and animals.

Fabrication and characterization of gum arabic- and maltodextrin-based microcapsules containing polyunsaturated oils.

BACKGROUND: Polyunsaturated oils have various health-promoting effects, however, they are highly prone to oxidation. Encapsulation using biopolymers is one of the most effective strategies to enhance oil stability. This research examined the potential of gum arabic and maltodextrin for microencapsulation of omega-3 rich oils, aiming to enhance encapsulation efficiency and stability of encapsulated oil. RESULTS: We encapsulated fish and flaxseed oils by emulsification-spray drying. Spray-dried microcapsules were prepared by oil-in-water emulsions consisting of 10 wt% oil and 30 wt% biopolymer (gum arabic, maltodextrin, or their mixture). Results showed that both microcapsules were spherical in shape with surface shrinkage, and exhibited amorphous structures. Gum arabic-based microcapsules had higher encapsulation efficiency as well as better storage stability for both types of oil. Flaxseed oil microcapsules generally had higher oxidative stability regardless of the type of wall material. CONCLUSIONS: Through a comprehensive characterization of the physical and chemical properties of the emulsions and resulting microcapsules, we proved gum arabic to be a more effective wall material for polyunsaturated oil microencapsulation, especially flaxseed oil. This study provides a promising approach to stabilize oils which are susceptible to deterioration, and facilitates their wider uses as food and nutraceutical products. © 2021 Society of Chemical Industry.

Therapeutic administration of Luteolin protects against Escherichia coli-derived Lipopolysaccharide-triggered inflammatory response and oxidative injury.

Endometritis reduces reproductive effectiveness and leads to significant financial losses in the dairy sector. Luteolin is a natural phyto-flavonoid compound with many biological activities. However, the therapeutic effect of Luteolin against lipopolysaccharides (LPS)-induced endometritis has not yet been explored. A total of eighty female Kunming mice were randomly assigned into four treatment groups (n = 20). Following a successful initiation of the endometritis model by LPS, Luteolin was intraperitoneally administered three times, at six-hour intervals between each injection in the Luteolin groups. The histopathological findings revealed that Luteolin significantly alleviated uterine injury induced by LPS. Moreover, Luteolin suppressed the synthesis of pro-inflammatory mediators [interleukin (IL)-1ß, IL-6, and tumor necrosis factor (TNF)-α] while promoting the synthesis of an anti-inflammatory mediator (IL-10) altered by LPS. Furthermore, Luteolin significantly mitigated the LPS-induced oxidative stress by scavenging malondialdehyde (MDA) and reactive oxygen species (ROS), accumulation and boosting the capacity of antioxidant enzyme activities such as superoxide dismutase 1 (SOD1), catalase (CAT), and glutathione peroxidase 1 (Gpx1) in the uterine tissue of mice. Additionally, injection of Luteolin markedly increased the expression of Toll-like receptors (TLR) 4 both at mRNA and protein levels under LPS stimulation. Western blotting and ELISA findings demonstrated that Luteolin suppressed the activation of the NF-κB pathway in response to LPS exposure in the uterine tissue of mice. Notably, Luteolin enhanced the anti-oxidant defense system by activating the Nrf2 signaling pathway under LPS exposure in the uterine tissue of mice. Conclusively, our findings demonstrated that Luteolin effectively alleviated LPS-induced endometritis via modulation of TLR4-associated Nrf2 and NF-κB signaling pathways.

EXPLORING the prophylactic potential of Azadirachta indica leaf extract against dyslipidemia.

ETHNOPHARMACOLOGICAL RELEVANCE: Several studies revealed that different parts of Azadirachta indica A. Juss, has therapeutic potential against inflammatory issues and dyslipidemia which is a major contributing cause to cardiovascular diseases, oxidative stress and serum glucose levels, etc. AIM OF STUDY: Present study was conducted to evaluate anti-dyslipidemic capacity of Azadirachta indica leaf extract in dyslipidemic rabbits. MATERIALS AND METHODS: Ethanolic extract of Azadirachta indica leaves was obtained by using Soxhlet apparatus. This extract was used for efficacy study on rabbits. In this context, 25 healthy rabbits were selected for study, Efficacy trial involved five groups of rabbits, 5 rabbits in each group; NC (Negative Control); healthy rabbits received normal diet. In remaining 20 rabbits, dyslipidemia was induced by using high fat diet for 28 days followed by administration of Azadirachta indica leaf ethanolic extract for 60 days in a dose-dependent manner. PC (Positive Control) include dyslipidemic rabbits received normal diet while G1, G2, G3 groups included dyslipidemic rabbits receiving different concentrations of Azadirachta indica leaf extract (i.e. 300, 500 and 700 mg/kg of body weight, respectively). Blood samples were analyzed for serum lipid profile after every 15 days to determine the effect of treatments. RESULTS: Significant reduction in total cholesterol (60 ± 3.4 mg/dL), triglycerides (40.31 ± 2.5 mg/dL) and low-density lipoprotein (28.87 ± 2.1 mg/dL) was observed in G2 (P ≤ 0.05)while a significant increase was observed in high-density lipoprotein (60.47 ± 1.7 mg/dL) of G2 (P ≤ 0.05) as compared to other groups. CONCLUSION: Results revealed that ethanolic extract of Azadirachta indica leaves in G2 group (@ 500 mg/kg of body weight) normalized lipid profile in dyslipidemic rabbits after 60 days of extract administration which significantly lowered TC, TG, LDL levels (P ≤ 0.05) and improved HDL level.

Compositional profiling and sensory analysis of cauliflower by-products-enriched muffins.

Cauliflower (Brassica oleracea var. botrytis) by-products (leaves, stems, stalks) (CBP) were successfully utilized in muffins as a model system and their feasibility of incorporation was investigated. CBP powder-based muffin formulations were made by the progressive replacement of wheat flour (WF) with 10%, 20%, and 30% of CBP. The physicochemical, pasting properties, antioxidant potential, textural characteristics, and sensorial attributes were analyzed. Substitution of CBP significantly (p < .05) resulted in an upsurge in crude protein, crude fiber, minerals, total phenolics, and total flavonoid contents, as well as total antioxidant activity values of muffins. The pasting properties were influenced by monitoring an increase in peak, breakdown, final, and setback viscosities. Although the addition of an increasing amount of CBP improved the nutritional characteristics, however, the increased level of replacement (>10%) had significant adverse effects on baking and physical characteristics. The specific loaf volume of the developed muffins decreased the crumb color which became darker, and enriched muffins were hardened in texture. Furthermore, sensory evaluation confirmed the positive effects of CBP incorporation only up to 10%. Overall, present results highlighted that supplementation of wheat muffins with 10% CBP is a beneficial approach to enrich them with nutrients and intensify their antioxidant potential.

Impact of high-intensity thermosonication treatment on spinach juice: Bioactive compounds, rheological, microbial, and enzymatic activities.

To study the impacts of thermosonication (TS), the spinach juice treated with TS (200 W, 400 W, and 600 W, 30 kHz, at 60 ± 1 °C for 20 mint) were investigated for bioactive compounds, antioxidant activities, color properties, particle size, rheological behavior, suspension stability, enzymatic and microbial loads. As a result, TS processing significantly improved the bioactive compounds (total flavonols, total flavonoids, total phenolic, carotenoids, chlorophyll, and anthocyanins), antioxidant activities (DPPH and FRAP assay) in spinach juice. Also, TS treatments had higher b*,L*, hue angle (h0), and chroma (C) values, while minimuma* value as compared to untreated and pasteurized samples. TS processing significantly reduced the particle size, improved the suspension stability and rheological properties (shear stress, apparent viscosity, and shear rate) of spinach juice as compared to the untreated and pasteurized sample. TS plays a synergistic part in microbial reduction and gained maximum microbial safety. Moreover, TS treatments inactivated the polyphenol oxidase and peroxidase from 0.97 and 0.034 Abs min-1 (untreated) to 0.31 and 0.018 Abs min-1, respectively. The spinach juice sample treated at a high intensity (600 W, 30 kHz, at 60 ± 1 °C for 20 mint, TS3) exhibited complete inactivation of microbial loads (<1 log CFU/ml), the highest reduction in enzymatic activities, better suspension stability, color properties, and highest bioactive compounds. Collectively, the verdicts proposed that TS processing could be a worthwhile option to pasteurize the spinach juice to enhance the overall quality.

Effect of Dietary Supplementation of Bioactive Peptides on Antioxidant Potential of Broiler Breast Meat and Physicochemical Characteristics of Nuggets.

Poultry meat is generally exposed to quality deterioration due to lipid oxidation during storage. Oxidative stability of meat can be increased by feed supplementation. Aim of the current study was to investigate the effect of dietary supplementation of fish waste derived bioactive peptides on antioxidant potential of broiler breast meat and physico-chemical characteristics and quality parameters of nuggets prepared from breast meat. 180 broiler birds (six groups of 30 birds) were purchased. Each group was given different concentrations of bioactive peptides i.e. 0, 50, 100, 150, 200, and 250 mg/kg feed. After completion of six weeks birds were slaughtered and breast meat was stored at -18°C for six months. Nuggets were prepared and stored at -18°C for 45 days. Meat samples were analyzed for antioxidant activity [total phenolic contents (TPC), DPPH• scavenging activity, and ferric reducing antioxidant power] and lipid oxidation assay at regular intervals of 1, 2, 3, 4, 5, and 6 months while nuggets were analyzed for quality (pH, color, texture and water holding capacity) parameters after regular interval of 15 days. A significant (p<0.05) effect of feed supplementation was observed on antioxidant status such as TPC, DPPH• scavenging activity, and FRAP of broiler breast meat. Dietary interventions of bioactive peptides significantly (p<0.05) delayed lipid oxidation of breast meat than control. All the quality parameters were also significantly affected due to dietary bioactive peptides and storage duration. Thus, dietary interventions of bioactive peptides can increase the antioxidant and shelf stability of broiler breast meat and nuggets.

Food protein aggregates as vitamin-matrix carriers: impact of processing conditions.

We studied the ability of protein aggregates for loading and protection of α-tocopherol, a model of heat- and light-sensitive bioactive compounds. Aqueous dispersions of whey proteins (4.5 wt.%, pH 6.7) in the absence and presence of α-tocopherol (4 wt.%) were prepared using an ultradisperser (10,000 rpm for 10 min and 65 °C), and then submitted to further high-pressure homogenisation (HPH) at 300 or 1200 bar for 12 cycles. Relative to free-vitamin dispersions, increasing HPH conditions in the presence of vitamin led to higher protein denaturation, more tryptophan quenching and wavelength blue-shift (by 10nm), in parallel with increased zeta potential values (by -10 mV), particle sizes (by 50%), and newly formed protein dimers, trimers and high molecular weight aggregates. As a result, the degree of vitamin degradation under increasing HPH and long-term storage was shown to decrease from 66% (ultradisper) to 50%, or to 30% (subject to further treatments at 300 or 1200 bar, respectively).

Lipid nanoparticles as vitamin matrix carriers in liquid food systems: On the role of high-pressure homogenisation, droplet size and adsorbed materials.

Twelve oil-in-water nano-emulsions were prepared using a melt high-pressure homogenisation process (HPH) at 300, 800 or 1200 bar. The resulting emulsions containing 20 wt% palm oil in the absence or presence of α-tocopherol were stabilised by whey proteins alone or in mixture with lecithin. Lipid nanoparticles in these emulsions were characterized for their particle size, surface charge and protein surface concentration (PSC) in relation to their stability against aggregation and coalescence, and to their ability for encapsulation and protection of α-tocopherol against chemical degradation. Increasing HPH values were accompanied by the formation of lipid nanoparticles with decreasing size and PSC, but increasing long-term stability against aggregation and coalescence in parallel with an increase in α-tocopherol degradation (up to 15 wt% for 1200 bar). Presence of α-tocopherol, led to increasing (or decreasing) PSC values with increasing (or decreasing) HPH values for lipid nanoparticles stabilised by proteins alone (or in mixture with lecithins). In addition to these structural properties, the ability for α-tocopherol long-term stability of nanoparticles in emulsions was shown to differ more depending on their adsorbed materials (protein alone, or in mixture with lecithin) than on their particle size values. After 2 months storage, α-tocopherol in emulsions prepared at 300, 800 or 1200 bar was protected against chemical degradation at 79, 77, 67 wt%, respectively, when whey proteins were used alone, instead of 66, 63, 48 wt% when proteins were used in mixture with lecithins. These results indicated the dominant role of adsorbed proteins on the protection of vitamin models by nanoemulsions. They are of a great technological importance for production of lipid nanoparticles presenting a high volume-to-diameter ratio values and consequently high exchange surfaces between the matrix carrier and water and oxygen environmental factors.