scholarly journals Biopolymer Extracted from Anadenanthera colubrina (Red Angico Gum) Exerts Therapeutic Potential in Mice: Antidiarrheal Activity and Safety Assessment

2020 ◽  
Vol 13 (1) ◽  
pp. 17 ◽  
Author(s):  
Thiago S. L. Araújo ◽  
Taiane M. de Oliveira ◽  
Nayara A. de Sousa ◽  
Luan K.M. Souza ◽  
Francisca B. M. Sousa ◽  
...  
Keyword(s):  
Therapeutic Potential ◽  
Gastrointestinal Transit ◽  
High Yield ◽  
Enzyme Linked Immunosorbent Assay ◽  
E Coli ◽  
Ft Ir ◽  
Antidiarrheal Activity ◽  
Pre Treatment ◽  
Smooth Muscle Contractions

Anadenanthera colubrina var. cebil (Griseb.) Altschul (Fabaceae family), commonly known as the red angico tree, is a medicinal plant found throughout Brazil’s semi-arid area. In this study, a chemical analysis was performed to investigate the antidiarrheal activity and safety profile of red angico gum (RAG), a biopolymer extracted from the trunk exudate of A. colubrina. Upon FT-IR spectroscopy, RAG showed bands in the regions of 1608 cm−1, 1368 cm−1, and 1029 cm−1, which relate to the vibration of O–H water molecules, deformation vibration of C-O bands, and vibration of the polysaccharide C-O band, respectively, all of which are relevant to glycosidic bonds. The peak molar mass of RAG was 1.89 × 105 g/mol, with the zeta potential indicating electronegativity. RAG demonstrated high yield and solubility with a low degree of impurity. Pre-treatment with RAG reduced the total diarrheal stool and enteropooling. RAG also enhanced Na+/K+-ATPase activity and reduced gastrointestinal transit, and thereby inhibited intestinal smooth muscle contractions. Enzyme-Linked Immunosorbent Assay (ELISA) demonstrated that RAG can interact with GM1 receptors and can also reduce E. coli-induced diarrhea in vivo. Moreover, RAG did not induce any signs of toxicity in mice. These results suggest that RAG is a possible candidate for the treatment of diarrheal diseases.

scholarly journals Antioxidant, analgesic and anti-inflammatory effects of lavender essential oil

2015 ◽  
Vol 87 (2 suppl) ◽  
pp. 1397-1408 ◽  
Author(s):  
GABRIELA L. DA SILVA ◽  
CAROLINA LUFT ◽  
ADROALDO LUNARDELLI ◽  
ROBSON H. AMARAL ◽  
DENIZAR A. DA SILVA MELO ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
Essential Oil ◽  
Therapeutic Potential ◽  
Positive Control ◽  
Croton Oil ◽  
Antinociceptive Effects ◽  
Anti Inflammatory ◽  
Pre Treatment ◽  
Dose Dependent

Several studies have investigated the antinociceptive, immunomodulatory and anti-inflammatory properties of compounds found in the lavender essential oil (LEO), however to date, there is still lack of substantial data. The objective of this study was to assess the antioxidant, anti-inflammatory and antinociceptive effects of lavender essential oil. The 1,1-diphenyl-2-picrylhydrazyl radical decolorization assay was used for antioxidant activity evaluation. The anti-inflammatory activity was tested using two models of acute inflammation: carrageenan-induced pleurisy and croton oil-induced ear edema. The antinociceptive activity was tested using the pain model induced by formalin. LEO has antioxidant activity, which is dose-dependent response. The inflammatory response evoked by carrageenan and by croton oil was reduced through the pre-treatment of animals with LEO. In the pleurisy model, the drug used as positive control, dexamethasone, was more efficacious. However, in the ear swelling, the antiedematogenic effect of the oil was similar to that observed for dexamethasone. In the formalin test, LEO consistently inhibited spontaneous nociception and presented a similar effect to that of tramadol. The results of this study reveal (in vivo) the analgesic and anti-inflammatory activities of LEO and demonstrates its important therapeutic potential.

scholarly journals Synthesis and Posttranslational Regulation of Pyruvate Formate-Lyase in Lactococcus lactis

2000 ◽  
Vol 182 (17) ◽  
pp. 4783-4788 ◽  
Author(s):  
Claus Rix Melchiorsen ◽  
Kirsten Væver Jokumsen ◽  
John Villadsen ◽  
Mads G. Johnsen ◽  
Hans Israelsen ◽  
...  
Keyword(s):  
Lactococcus Lactis ◽  
Protein Level ◽  
Anaerobic Growth ◽  
Enzyme Linked Immunosorbent Assay ◽  
Posttranslational Regulation ◽  
Aerobic Growth ◽  
Acid Product ◽  
Pyruvate Formate Lyase ◽  
E Coli

ABSTRACT The enzyme pyruvate formate-lyase (PFL) from Lactococcus lactis was produced in Escherichia coli and purified to obtain anti-PFL antibodies that were shown to be specific forL. lactis PFL. It was demonstrated that activated L. lactis PFL was sensitive to oxygen, as in E. coli, resulting in the cleavage of the PFL polypeptide. The PFL protein level and its in vivo activity and regulation were shown by Western blotting, enzyme-linked immunosorbent assay, and metabolite measurement to be dependent on the growth conditions. The PFL level during anaerobic growth on the slowly fermentable sugar galactose was higher than that on glucose. This shows that variation in the PFL protein level may play an important role in the regulation of metabolic shift from homolactic to mixed-acid product formation, observed during growth on glucose and galactose, respectively. During anaerobic growth in defined medium, complete activation of PFL was observed. Strikingly, although no formate was produced during aerobic growth of L. lactis, PFL protein was indeed detected under these conditions, in which the enzyme is dispensable due to the irreversible inactivation of PFL by oxygen. In contrast, no oxygenolytic cleavage was detected during aerobic growth in complex medium. This observation may be the result of either an effective PFL deactivase activity or the lack of PFL activation. In E. coli, the PFL deactivase activity resides in the multifunctional alcohol dehydrogenase ADHE. It was shown that inL. lactis, ADHE does not participate in the protection of PFL against oxygen under the conditions analyzed. Our results provide evidence for major differences in the mechanisms of posttranslational regulation of PFL activity in E. coli and L. lactis.

scholarly journals Reliable method for high quality His-tagged and untagged E. coli phosphoribosyl phosphate synthase (Prs) purification

2019 ◽  
Author(s):  
Walter Beata Maria ◽  
Szulc Aneta ◽  
Glinkowska Monika
Keyword(s):  
Quaternary Structure ◽  
Physiological State ◽  
Building Blocks ◽  
High Yield ◽  
Phosphoribosyl Pyrophosphate ◽  
Protein Variant ◽  
E Coli ◽  
Nickel Affinity Chromatography

ABSTRACTPrs (phosphoribosyl pyrophosphate synthase) is a broadly conserved protein that synthesises 5-phosphoribosyl 1-pyrophospate (PRPP); a substrate for biosynthesis of at least 10 enzymatic pathways including biosynthesis of DNA building blocks – purines and pyrimidines. In Escherichia coli, it is a protein of homo-hexameric quaternary structure, which can be challenging to work with, due to frequent aggregation and activity loss. Several studies showed brief purification protocols for various bacterial PRPP synthases, in most cases involving ammonium sulfate precipitation.Here, we provide a protocol for expression of E. coli Prs protein in Rosetta (DE3) and BL21 (DE3) pLysE strains and a detailed method for His-Prs and untagged Prs purification on nickel affinity chromatography columns. This protocol allows purification of proteins with high yield, purity and activity. We report here N-terminally His-tagged protein fusions, stable and active, providing that the temperature around 20 °C is maintained at all stages, including centrifugation. Moreover, we successfully applied this method to purify two enzyme variants with K194A and G9S alterations. The K194A mutation in conserved lysine residue results in protein variant unable to synthetize PRPP, while the G9S alteration originates from prs-2 allele variant which was previously related to thermo-sensitive growth. His-PrsG9S protein purified here, exhibited comparable activity as previously observed in-vivo suggesting the proteins purified with our protocol resemble their physiological state.The protocol for Prs purification showed here indicates guidance to improve stability and quality of the protein and to ensure more reliable results in further assays in-vitro.

The Neuroprotective Effect of Picroside II from Hu-Huang-Lian against Oxidative Stress

2007 ◽  
Vol 35 (04) ◽  
pp. 681-691 ◽  
Author(s):  
Ting Li ◽  
Jian-Wen Liu ◽  
Xiao-Dong Zhang ◽  
Ming-Chuan Guo ◽  
Guang Ji
Keyword(s):  
Oxidative Stress ◽  
Pc12 Cells ◽  
Therapeutic Potential ◽  
Neuroprotective Effect ◽  
Active Constituent ◽  
Sod Activity ◽  
Picroside Ii ◽  
Pre Treatment

Picroside II is an active constituent extracted from the traditional Chinese medicine (TCM) Hu-Huang-Lian. To evaluate the neuroprotective effect of picroside II, PC12 cells were treated with glutamate in vitro and male ICR mice were treated with AlCl 3in vivo. Pre-treatment of PC12 cells with picroside II could enhance the cell viability and decrease the level of intracellular reactive oxygen species (ROS) induced by glutamate. By DNA fragmentation and flow cytometry assay, picroside II (1.2 mg/ml) significantly prevented glutamate-induced cell apoptosis. In the animal study, amnesia was induced in mice by AlCl 3 (100 mg/kg/d, i.v.). Pricroside II, at the dose of 20 and 40 mg/kg/d (i.g.), markedly ameliorated AlCl 3-induced learning and memory dysfunctions and attenuated AlCl 3-induced histological changes. This was associated with the significant increased superoxide dismutase (SOD) activity in the brain of experimental mice. All these results indicated that picroside II possessed the therapeutic potential in protecting against neurological injuries damaged by oxidative stress.

scholarly journals Leukocyte Production of Inflammatory Mediators Is Inhibited by the Antioxidants Phloretin, Silymarin, Hesperetin, and Resveratrol

2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
Jezrom B. Fordham ◽  
Afsar Raza Naqvi ◽  
Salvador Nares
Keyword(s):  
Inflammatory Mediators ◽  
Mononuclear Cells ◽  
Therapeutic Potential ◽  
Aggregatibacter Actinomycetemcomitans ◽  
Significant Inhibition ◽  
Periodontal Pathogen ◽  
Enzyme Linked Immunosorbent Assay ◽  
Peripheral Blood Mononuclear ◽  
Differentiation Factors

Antioxidants possess significant therapeutic potential for the treatment of inflammatory disorders. One such disorder is periodontitis characterised by an antimicrobial immune response, inflammation, and irreversible changes to the supporting structures of the teeth. Recognition of conserved pathogen-associated molecular patterns is a crucial component of innate immunity to Gram-negative bacteria such asEscherichia coli, as well as the periodontal pathogenAggregatibacter actinomycetemcomitans. In this study, we investigated the antioxidants Phloretin, Silymarin, Hesperetin, and Resveratrol to ascertain whether they altered the production of inflammatory mediators by innately-activated leukocytes. Peripheral blood mononuclear cells were stimulated with lipopolysaccharide purified fromAggregatibacter actinomycetemcomitans, and the production of cytokines, chemokines, and differentiation factors was assayed by enzyme-linked immunosorbent assay, cytometric bead array, and RT-PCR. Significant inhibition of these factors was achieved upon treatment with Phloretin, Silymarin, Hesperetin, and Resveratrol. These data further characterise the potent anti-inflammatory properties of antioxidants. Their ability to inhibit the production of inflammatory cytokines, chemokines, and differentiation factors by a heterogeneous population of leukocytes has clear implications for their therapeutic potentialin vivo.

scholarly journals Carnosine Protects Macrophages against the Toxicity of Aβ1-42 Oligomers by Decreasing Oxidative Stress

Biomedicines ◽  
2021 ◽  
Vol 9 (5) ◽  
pp. 477
Author(s):  
Giuseppe Caruso ◽  
Cristina Benatti ◽  
Nicolò Musso ◽  
Claudia G. Fresta ◽  
Annamaria Fidilio ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Therapeutic Potential ◽  
Radical Scavenging ◽  
Antioxidant Systems ◽  
Trypan Blue Exclusion ◽  
New Findings ◽  
Pre Treatment ◽  
The Brain

Carnosine (β-alanyl-L-histidine) is a naturally occurring endogenous peptide widely distributed in excitable tissues such as the brain. This dipeptide has well-known antioxidant, anti-inflammatory, and anti-aggregation activities, and it may be useful for treatment of neurodegenerative disorders such as Alzheimer’s disease (AD). In this disease, peripheral infiltrating macrophages play a substantial role in the clearance of amyloid beta (Aβ) peptides from the brain. Correspondingly, in patients suffering from AD, defects in the capacity of peripheral macrophages to engulf Aβ have been reported. The effects of carnosine on macrophages and oxidative stress associated with AD are consequently of substantial interest for drug discovery in this field. In the present work, a model of stress induced by Aβ1-42 oligomers was investigated using a combination of methods including trypan blue exclusion, microchip electrophoresis with laser-induced fluorescence, flow cytometry, fluorescence microscopy, and high-throughput quantitative real-time PCR. These assays were used to assess the ability of carnosine to protect macrophage cells, modulate oxidative stress, and profile the expression of genes related to inflammation and pro- and antioxidant systems. We found that pre-treatment of RAW 264.7 macrophages with carnosine counteracted cell death and apoptosis induced by Aβ1-42 oligomers by decreasing oxidative stress as measured by levels of intracellular nitric oxide (NO)/reactive oxygen species (ROS) and production of peroxynitrite. This protective activity of carnosine was not mediated by modulation of the canonical inflammatory pathway but instead can be explained by the well-known antioxidant and free-radical scavenging activities of carnosine, enhanced macrophage phagocytic activity, and the rescue of fractalkine receptor CX3CR1. These new findings obtained with macrophages challenged with Aβ1-42 oligomers, along with the well-known multimodal mechanism of action of carnosine in vitro and in vivo, substantiate the therapeutic potential of this dipeptide in the context of AD pathology.

scholarly journals Immunoreactive Proteins of Dormant Mycobacterium tuberculosis Cells

2021 ◽  
Vol 57 (2) ◽  
pp. 170-179
Author(s):  
K. A. Trutneva ◽  
V. G. Avdienko ◽  
G. R. Demina ◽  
M. O. Shleeva ◽  
M. S. Shumkov ◽  
...  
Keyword(s):  
Protein Profile ◽  
Enzyme Linked Immunosorbent Assay ◽  
2D Electrophoresis ◽  
E Coli ◽  
Dormant Cells ◽  
Healthy Donors ◽  
Antigenic Proteins ◽  
Latent Tb

Abstract The protein profile of dormant Mtb obtained after the gradual acidification of Mtb culture was studied to find antigenic proteins for humans that are expressed by M. tuberculosis (Mtb) cells in vitro under conditions close to the situation of persistence in vivo. According to 2D electrophoresis, a significant diversity of proteins in dormant cells was found. However, the representation of individual proteins in dormant versus active cells differed substantially. Immunoblotting in different protein fractions of dormant cells revealed ten proteins that are able to bind antibodies in pooled sera of TB patients. Two proteins (Rv2018 and Rv0341) are new immunogenics that were not previously found in other studies. Four proteins (Rv0341, Rv2018, Rv1509, Rv2986) with the maximal structural specificity for Mtb due to their unique extended domains were selected for further analysis. These proteins were expressed in E. coli cells and studied via enzyme-linked immunosorbent assay (ELISA) for the immunogenicity of individual sera of TB patients and healthy donors. All proteins were found to have the ability to react with individual sera of TB patients. In TB patients, 5–45% (depending on the particulate protein) have a titer that is higher than the average titers of healthy donors +SD; the most immunogenic was protein Rv2986. Thus, the application of phenotypically changed (dormant) Mtb cells makes it possible to identify a specific repertoire of immunodominant proteins that could be used in the construction of polypeptides that are useful for the serodiagnosis of active/latent TB.

scholarly journals Novel NFκB Inhibitor SC75741 Mitigates Chondrocyte Degradation and Prevents Activated Fibroblast Transformation by Modulating miR-21/GDF-5/SOX5 Signaling

2021 ◽  
Vol 22 (20) ◽  
pp. 11082
Author(s):  
Pei-Wei Weng ◽  
Vijesh Kumar Yadav ◽  
Narpati Wesa Pikatan ◽  
Iat-Hang Fong ◽  
I-Hsin Lin ◽  
...  
Keyword(s):  
Therapeutic Potential ◽  
Protective Role ◽  
Enzyme Linked Immunosorbent Assay ◽  
Molecule Inhibitor ◽  
Tnf Α ◽  
Inflammatory Profile ◽  
Fibroblast Transformation ◽  
Tgf Β1

Osteoarthritis (OA) is a common articular disease manifested by the destruction of cartilage and compromised chondrogenesis in the aging population, with chronic inflammation of synovium, which drives OA progression. Importantly, the activated synovial fibroblast (AF) within the synovium facilitates OA through modulating key molecules, including regulatory microRNAs (miR’s). To understand OA associated pathways, in vitro co-culture system, and in vivo papain-induced OA model were applied for this study. The expression of key inflammatory markers both in tissue and blood plasma were examined by qRT-PCR, western blot, immunohistochemistry, enzyme-linked immunosorbent assay (ELISA) and immunofluorescence assays. Herein, our result demonstrated, AF-activated human chondrocytes (AC) exhibit elevated NFκB, TNF-α, IL-6, and miR-21 expression as compared to healthy chondrocytes (HC). Importantly, AC induced the apoptosis of HC and inhibited the expression of chondrogenesis inducers, SOX5, TGF-β1, and GDF-5. NFκB is a key inflammatory transcription factor elevated in OA. Therefore, SC75741 (an NFκB inhibitor) therapeutic effect was explored. SC75741 inhibits inflammatory profile, protects AC-educated HC from apoptosis, and inhibits miR-21 expression, which results in the induced expression of GDF-5, SOX5, TGF-β1, BMPR2, and COL4A1. Moreover, ectopic miR-21 expression in fibroblast-like activated chondrocytes promoted osteoblast-mediated differentiation of osteoclasts in RW264.7 cells. Interestingly, in vivo study demonstrated SC75741 protective role, in controlling the destruction of the articular joint, through NFκB, TNF-α, IL-6, and miR-21 inhibition, and inducing GDF-5, SOX5, TGF-β1, BMPR2, and COL4A1 expression. Our study demonstrated the role of NFκB/miR-21 axis in OA progression, and SC75741′s therapeutic potential as a small-molecule inhibitor of miR-21/NFκB-driven OA progression.

Uncatalyzed Synthesis of New Antibacterial Bisarylidene Meldrum's Acid Derivatives Functionalized with Ether Groups

2019 ◽  
Vol 16 (10) ◽  
pp. 818-824
Author(s):  
Elham Moosazadeh ◽  
Enayatollah Sheikhhosseini ◽  
Dadkhoda Ghazanfari ◽  
Shahla Soltaninejad
Keyword(s):  
Staphylococcus Aureus ◽  
Mass Spectra ◽  
Condensation Reaction ◽  
Antibacterial Activities ◽  
High Yield ◽  
Meldrum's Acid ◽  
Meldrum’S Acid ◽  
E Coli ◽  
Ft Ir ◽  
Meldrum's Acid Derivatives

A series of new bisarylidene Meldrum’s acid derivatives (3a-i) were prepared in high yield by a condensation reaction between Meldrum's acid and ether functionalized dibenzaldehyde, without any catalyst. Regardless of the nature of the substitution, all the reactions were completed within 2-3 hours in ethanol at reflux condition. In the reactions, the column purification of the products was not needed. The FT-IR, 1H-NMR, 13C-NMR and mass spectra confirm the structure of the products. Furthermore, the antibacterial activities of some synthesized compounds were investigated. According to the results, these compounds showed good activities against Staphylococcus aureus, Bacillus cereus, E. coli and Pseudomonas aeruginosa.

scholarly journals Evaluation of In Vivo Antidiarrheal Activities of Hydroalcoholic Leaf Extract of Dodonaea viscosa L.(Sapindaceae) in Swiss Albino Mice

2019 ◽  
Vol 24 ◽  
pp. 2515690X1989195
Author(s):  
Jemal Abdela
Keyword(s):  
Plant Extract ◽  
Castor Oil ◽  
Leaf Extract ◽  
Gastrointestinal Transit ◽  
Control Group ◽  
Dodonaea Viscosa ◽  
Swiss Albino Mice ◽  
Antidiarrheal Activity ◽  
Albino Mice

Traditionally people used Dodonaea viscosa for the treatment of various ailments, including diarrhea. Therefore, this study was aimed to evaluate the antidiarrheal activity of the 80% methanolic leaf extract of D viscosa against castor oil-induced diarrhea in mice models. Different doses of 80% methanolic leaf extract of D viscosa (100, 200, and 400 mg/kg) were evaluated for their antidiarrheal activities using castor oil–induced diarrhea, gastrointestinal transit, and enteropooling models in Swiss albino mice. At all test doses, the plant extract showed significant ( P < .05) inhibition in the frequency of defecation of wet feces and total fecal output as compared to the control group. Similarly, at all dose ranges used the plant extract demonstrated significant ( P < .05) reduction in an intraluminal fluid accumulation as compared to the untreated group. Besides, at higher doses, the plant extract also indicated significant ( P < .05) antimotility activity in comparison with the control. In conclusion, these findings illustrated that the 80% methanolic leaf extract of D viscosa supported the traditional claim of antidiarrheal activity of the plant though further investigations are warranted.

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