A Simple and a Reliable Method to Quantify Antioxidant Activity In Vivo

The characterization of compounds with antioxidant activity is of great interest due to their ability to reduce reactive oxygen species production and, therefore, prevent some age-related diseases. Its antioxidant capacity can be analyzed by different methods both in vitro and in vivo. Caenorhabditis elegans is an in vivo model widely used in ageing research. Until now, available tests analyze functional effects in the worms, so the antioxidant activity of the compound is indirectly monitored. We have developed a simple and a reliable method to quantify internal antioxidant activity in vivo. To validate this method, we analyzed an aqueous green tea extract and two other compounds with a well-known antioxidant activity and without this activity. The results obtained (EC50 green tea = 21.76 ± 1.28 µg/mL; EC50 positive control = 8.50 ± 0.33 µg/mL; negative control EC50 > 500 µg/mL) can help in the design of further in vivo experiments. Thus, our method can be used as a previous screening capable of reducing the gap between in vitro and in vivo assays.

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LOWERING URIC ACID EFFICACY TEST OF THE COMBINED EXTRACT OF UNCARIA GAMBIR (HUNTER) ROXB. AND CAESALPINIA SAPPAN L. IN VIVO AND IN VITRO

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2018.v11i8.26020 ◽

2018 ◽

Vol 11 (8) ◽

pp. 166

Author(s):

Sri Ningsih ◽

Fahri Fahrudin

Keyword(s):

Antioxidant Activity ◽

Uric Acid ◽

Inhibitory Activity ◽

Positive Control ◽

Polyphenol Content ◽

Caesalpinia Sappan ◽

In Vivo Experiments ◽

A Value

Objective: Hyperuricemia (high uric acid levels) prevalence increased year by year. This study was aimed to elaborate the in vitro xanthine oxidase (XO) inhibitory activity and in vivo lowering hyperuricemic effect of Uncaria gambir (Hunter) Roxb) (gambir), Caesalpinia sappan L. (secang) and the combined extract of secang and gambir (formulae extract [FE]).Methods: Gambir and secang extracts were prepared by maceration with ethanol and FE was the proportioned combination of these two extracts. XO inhibitory activity was determined by measuring the formation of uric acid in the xanthine/XO system in vitro using allopurinol as a positive control at 100 ug/mL. Antioxidant activity was by 1,1-diphenyl-2-picrylhydrazyl radical reducing methods. The in vivo experiments were conducted in the oxonate-induced hyperuricemia rat model, in which FE was gavaged p.o. at the arrange dose of 75, 150, and 300 mg/kg bw for 2 weeks. Polyphenol content was measured using Folin–Ciocalteu reagent spectrophotometrically.Results: The XO inhibitory activity of FE was 80% of allopurinol, while secang and gambir were 98% and 50%, respectively. The strength was appropriate to the total polyphenol content, in which it decreased in the order of secang (99%) > FE (86%) > gambir (46%). Furthermore, FE at all tested doses was able to decrease uric acid levels. FE also demonstrated antioxidant activity with a value of 74% relative to Vitamin C at 4 ug/mL.Conclusion: These studies could be concluded that FE exhibited the ability to decrease uric acid level so that it was potential to be developed further as a uric acid-lowering agent.

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Regorafenib-Attenuated, Bleomycin-Induced Pulmonary Fibrosis by Inhibiting the TGF-β1 Signaling Pathway

International Journal of Molecular Sciences ◽

10.3390/ijms22041985 ◽

2021 ◽

Vol 22 (4) ◽

pp. 1985

Author(s):

Xiaohe Li ◽

Ling Ma ◽

Kai Huang ◽

Yuli Wei ◽

Shida Long ◽

...

Keyword(s):

Pulmonary Fibrosis ◽

Signaling Pathway ◽

Transforming Growth Factor ◽

Kinase Inhibitor ◽

In Vivo Experiments ◽

Age Related ◽

And Migration ◽

Tgf Β1

Idiopathic pulmonary fibrosis (IPF) is a fatal and age-related pulmonary disease. Nintedanib is a receptor tyrosine kinase inhibitor, and one of the only two listed drugs against IPF. Regorafenib is a novel, orally active, multi-kinase inhibitor that has similar targets to nintedanib and is applied to treat colorectal cancer and gastrointestinal stromal tumors in patients. In this study, we first identified that regorafenib could alleviate bleomycin-induced pulmonary fibrosis in mice. The in vivo experiments indicated that regorafenib suppresses collagen accumulation and myofibroblast activation. Further in vitro mechanism studies showed that regorafenib inhibits the activation and migration of myofibroblasts and extracellular matrix production, mainly through suppressing the transforming growth factor (TGF)-β1/Smad and non-Smad signaling pathways. In vitro studies have also indicated that regorafenib could augment autophagy in myofibroblasts by suppressing TGF-β1/mTOR (mechanistic target of rapamycin) signaling, and could promote apoptosis in myofibroblasts. In conclusion, regorafenib attenuates bleomycin-induced pulmonary fibrosis by suppressing the TGF-β1 signaling pathway.

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Effects of Food Processing on In Vivo Antioxidant and Hepatoprotective Properties of Green Tea Extracts

Antioxidants ◽

10.3390/antiox8120572 ◽

2019 ◽

Vol 8 (12) ◽

pp. 572 ◽

Cited By ~ 2

Author(s):

Xiao-Yu Xu ◽

Jie Zheng ◽

Jin-Ming Meng ◽

Ren-You Gan ◽

Qian-Qian Mao ◽

...

Keyword(s):

Green Tea ◽

Food Processing ◽

Antioxidant Activities ◽

Ultrasound Treatment ◽

Positive Effects ◽

In Vivo Experiments ◽

Study Results ◽

Negative Effect

Food processing can affect the nutrition and safety of foods. A previous study showed that tannase and ultrasound treatment could significantly increase the antioxidant activities of green tea extracts according to in vitro evaluation methods. Since the results from in vitro and in vivo experiments may be inconsistent, the in vivo antioxidant activities of the extracts were studied using a mouse model of alcohol-induced acute liver injury in this study. Results showed that all the extracts decreased the levels of aspartate transaminase and alanine aminotransferase in serum, reduced the levels of malondialdehyde and triacylglycerol in the liver, and increased the levels of catalase and glutathione in the liver, which can alleviate hepatic oxidative injury. In addition, the differences between treated and original extracts were not significant in vivo. In some cases, the food processing can have a negative effect on in vivo antioxidant activities. That is, although tannase and ultrasound treatment can significantly increase the antioxidant activities of green tea extracts in vitro, it cannot improve the in vivo antioxidant activities, which indicates that some food processing might not always have positive effects on products for human benefits.

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Cleaved CD31 as a target for in vivo molecular imaging of inflammation

Scientific Reports ◽

10.1038/s41598-019-56163-x ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 1

Author(s):

Jonathan Vigne ◽

Sylvie Bay ◽

Rachida Aid-Launais ◽

Guillaume Pariscoat ◽

Guillaume Rucher ◽

...

Keyword(s):

Molecular Imaging ◽

Molecular Form ◽

Positive Control ◽

Negative Control ◽

Stability Study ◽

Biodistribution Studies ◽

Wide Range ◽

Significant Difference

AbstractThere is a need for new targets to specifically localize inflammatory foci, usable in a wide range of organs. Here, we hypothesized that the cleaved molecular form of CD31 is a suitable target for molecular imaging of inflammation. We evaluated a bioconjugate of D-P8RI, a synthetic peptide that binds all cells with cleaved CD31, in an experimental rat model of sterile acute inflammation. Male Wistar rats were injected with turpentine oil into the gastrocnemius muscle two days before 99mTc-HYNIC-D-P8RI (or its analogue with L-Proline) SPECT/CT or [18F]FDG PET/MRI. Biodistribution, stability study, histology, imaging and autoradiography of 99mTc-HYNIC-D-P8RI were further performed. Biodistribution studies revealed rapid elimination of 99mTc-HYNIC-D-P8RI through renal excretion with almost no uptake from most organs and excellent in vitro and in vivo stability were observed. SPECT/CT imaging showed a significant higher 99mTc-HYNIC-D-P8RI uptake compared with its analogue with L-Proline (negative control) and no significant difference compared with [18F]FDG (positive control). Moreover, autoradiography and histology revealed a co-localization between 99mTc-HYNIC-D-P8RI uptake and inflammatory cell infiltration. 99mTc-HYNIC-D-P8RI constitutes a new tool for the detection and localization of inflammatory sites. Our work suggests that targeting cleaved CD31 is an attractive strategy for the specific in vivo imaging of inflammatory processes.

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Brazilian Red Propolis Is as Effective as Amoxicillin in Controlling Red-Complex of Multispecies Subgingival Mature Biofilm In Vitro

Antibiotics ◽

10.3390/antibiotics9080432 ◽

2020 ◽

Vol 9 (8) ◽

pp. 432

Author(s):

Kadmo Azevedo de Figueiredo ◽

Helio Doyle Pereira da Silva ◽

Stela Lima Farias Miranda ◽

Francisco Jerfeson dos Santos Gonçalves ◽

Arlene Pereira de Sousa ◽

...

Keyword(s):

Metabolic Activity ◽

Positive Control ◽

Negative Control ◽

In Vivo Studies ◽

Complex Species ◽

Hybridization Data ◽

Cell Counts ◽

Actinomyces Species

This study investigated the effects of Brazilian Red Propolis (BRP) extract on seven-day-old multispecies subgingival biofilms. Mixed biofilm cultures containing 31 species associated with periodontal health or disease were grown for six days on a Calgary device. Then, mature biofilms were treated for 24 h with BRP extract at different concentrations (200–1600 µg/mL), amoxicillin (AMOXI) at 54 µg/mL (positive control) or vehicle (negative control). Biofilm metabolic activity was determined by colorimetry, and bacterial counts/proportions were determined by DNA–DNA hybridization. Data were analyzed by Kruskal–Wallis and Dunn’s tests. Treatment with BRP at 1600, 800 and 400 μg/mL reduced biofilm metabolic activity by 56%, 56% and 57%, respectively, as compared to 65% reduction obtained with AMOXI. Mean total cell counts were significantly reduced in all test groups (~50–55%). Lower proportions of red, green and yellow complex species were observed upon treatment with BRP (400 µg/mL) and AMOXI, but only AMOXI reduced the proportions of Actinomyces species. In conclusion, BRP extract was as effective as AMOXI in killing seven-day-old multispecies biofilm pathogens and did not affect the levels of the host-compatible Actinomyces species. These data suggest that BRP may be an alternative to AMOXI as an adjunct in periodontal therapy. In vivo studies are needed to validate these results.

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In vivo activities of ceftriaxone and vancomycin against Borrelia spp. in the mouse brain and other sites.

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.40.11.2632 ◽

1996 ◽

Vol 40 (11) ◽

pp. 2632-2636 ◽

Cited By ~ 28

Author(s):

R J Kazragis ◽

L L Dever ◽

J H Jorgensen ◽

A G Barbour

Keyword(s):

Body Weight ◽

Lyme Disease ◽

Positive Control ◽

Negative Control ◽

Scid Mice ◽

Immunodeficient Mice ◽

Infected Adult ◽

The Brain

Borrelia burgdorferi, the agent of Lyme disease, and B. turicatae, a neurotropic agent of relapsing fever, are susceptible to vancomycin in vitro, with an MIC of 0.5 microgram/ml. To determine the activity of vancomycin in vivo, particularly in the brain, we infected adult immunocompetent BALB/c and immunodeficient CB-17 scid mice with B. burgdorferi or B. turicatae. The mice were then treated with vancomycin, ceftriaxone as a positive control, or normal saline as a negative control. The effectiveness of treatment was assessed by cultures of blood and brain and other tissues. Ceftriaxone at a dose of 25 mg/kg of body weight administered every 12 h for 7 to 10 days eliminated cultivable B. burgdorferi or B. turicatae from all BALB/c or scid mice in the study. Vancomycin at 30 mg/kg administered every 12 h was effective in eliminating infection from immunodeficient mice if treatment was started within 3 days of the onset of infection. If treatment with vancomycin was delayed for 7 days or more, vancomycin failed to eradicate infection with B. burgdorferi or B. turicatae from immunodeficient mice. The failure of vancomycin in eradicating established infections in immunodeficient mice was associated with the persistence of viable spirochetes in the brain during antibiotic treatment.

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Chloroquine enhances temozolomide cytotoxicity in malignant gliomas by blocking autophagy

Neurosurgical FOCUS ◽

10.3171/2014.9.focus14504 ◽

2014 ◽

Vol 37 (6) ◽

pp. E12 ◽

Cited By ~ 63

Author(s):

Encouse B. Golden ◽

Hee-Yeon Cho ◽

Ardeshir Jahanian ◽

Florence M. Hofman ◽

Stan G. Louie ◽

...

Keyword(s):

Molecular Mechanisms ◽

Malignant Gliomas ◽

Glioma Cells ◽

In Vivo Model ◽

Autophagosome Formation ◽

Individual Drug ◽

In Vivo Experiments ◽

Associated Proteins

Object In a recent clinical trial, patients with newly diagnosed glioblastoma multiforme benefited from chloroquine (CQ) in combination with conventional therapy (resection, temozolomide [TMZ], and radiation therapy). In the present study, the authors report the mechanism by which CQ enhances the therapeutic efficacy of TMZ to aid future studies aimed at improving this therapeutic regimen. Methods Using in vitro and in vivo experiments, the authors determined the mechanism by which CQ enhances TMZ cytotoxicity. They focused on the inhibition-of-autophagy mechanism of CQ by knockdown of the autophagy-associated proteins or treatment with autophagy inhibitors. This mechanism was tested using an in vivo model with subcutaneously implanted U87MG tumors from mice treated with CQ in combination with TMZ. Results Knockdown of the autophagy-associated proteins (GRP78 and Beclin) or treatment with the autophagy inhibitor, 3-methyl adenine (3-MA), blocked autophagosome formation and reduced CQ cytotoxicity, suggesting that autophagosome accumulation precedes CQ-induced cell death. In contrast, blocking autophagosome formation with knockdown of GRP78 or treatment with 3-MA enhanced TMZ cytotoxicity, suggesting that the autophagy pathway protects from TMZ-induced cytotoxicity. CQ in combination with TMZ significantly increased the amounts of LC3B-II (a marker for autophagosome levels), CHOP/GADD-153, and cleaved PARP (a marker for apoptosis) over those with untreated or individual drug-treated glioma cells. These molecular mechanisms seemed to take place in vivo as well. Subcutaneously implanted U87MG tumors from mice treated with CQ in combination with TMZ displayed higher levels of CHOP/GADD-153 than did untreated or individual drug-treated tumors. Conclusions Taken together, these results demonstrate that CQ blocks autophagy and triggers endoplasmic reticulum stress, thereby increasing the chemosensitivity of glioma cells to TMZ.

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Combination of garlic - shatterstone herb powder to control Streptococcus agalactiae infection in tilapia

Jurnal Akuakultur Indonesia ◽

10.19027/jai.14.79-89 ◽

2015 ◽

Vol 14 (1) ◽

pp. 79

Author(s):

Ririn Nurul Fauziah ◽

Dinamella Wahjuningrum ◽

, Sukenda ◽

, Ranta

Keyword(s):

Growth Rate ◽

Streptococcus Agalactiae ◽

Allium Sativum ◽

Positive Control ◽

Quality Parameters ◽

Negative Control ◽

Fish Pathogen ◽

Phyllanthus Niruri

ABSTRACT This study was aimed at determining potential of combination powder of garlic Allium sativum-shatterstone herb Phyllanthus niruri supplemented in feed against S. agalactiae infection in tilapia. Four concentrations of combination powder of A. sativum-P. Niruri; 20+5, 20+10, 20+15 and 20+20 ppt respectively were investigated for their ability to inhibit bacterial fish pathogen. Combination dose of 20+15 ppt produced the highest inhibitory zones in in vitro test. In vivo test consisted of three treatments with three replications, namely positive control (K+), negative control (K-) and the treatment of A. sativum-P. niruri suplemented in feed (BM). The test perfomed on tilapia with weight of 10.33 ± 1.63 g and were reared at density of 10 ind/aquarium. The fish was fed for 14 days, then injected intraperitoneally with 0.1 mL S. agalactiae at concentration of 105 cfu/mL for positive control and BM groups. Survival, growth rate, feed response, hematological and water quality parameters were observed for 10 days. This study showed that the suplemented-feed-fish (BM) showed better growth rate, feed response, and survival (83.3%) than positive control (36.7%) at P<0.05. In addition, A. sativum-P. niruri suplemented in feed was also able to enhance the immune response by increasing phagocytic activity. Keywords: Streptococcus agalactiae, phytopharmacy, Allium sativum-Phyllanthus niruri, tilapia ABSTRAK Penelitian ini bertujuan untuk menganalisis potensi campuran tepung bawang putih Allium sativum-meniran Phyllanthus niruri dalam pakan terhadap pencegahan infeksi bakteri S. agalactiae pada ikan nila. Empat konsentrasi campuran tepung bawang putih-meniran yaitu 20+5 ppt, 20+10 ppt, 20+15 ppt dan 20+20 ppt masing-masing diuji kemampuannya dalam menghambat bakteri patogen pada ikan. Campuran dosis 20+15 ppt menghasilkan zona hambat terbaik dalam uji in vitro. Uji in vivo terdiri atas tiga perlakuan dengan tiga ulangan yaitu kontrol positif, kontrol negatif, dan perlakuan pakan yang mengandung bawang putih-meniran (BM). Uji ini dilakukan pada ikan nila berbobot 10,33±1,63 g yang dipelihara di akuarium dengan kepadatan 10 ekor/akuarium. Ikan diberi pakan perlakuan selama 14 hari kemudian diinjeksi secara intraperitoneal dengan bakteri S. agalactiae sebanyak 0,1 mL dengan kepadatan 105 cfu/mL pada perlakuan kontrol positif dan perlakuan BM. Parameter kelangsungan hidup, laju pertumbuhan, respons pakan, parameter hematologi, dan kualitas air diamati selama sepuluh hari. Hasil dari penelitian ini menunjukkan bahwa pemberian BM dalam pakan memberikan laju pertumbuhan, respons pakan, dan sintasan (83,3%) yang lebih baik daripada kontrol positif (36,7%) pada P<0,05. Pakan yang mengandung campuran bawang putih-meniran ini juga mampu meningkatkan respons imun dengan adanya peningkatan aktivitas fagositosis. Kata kunci: Streptococcus agalactiae, fitofarmaka, Allium sativum-Phyllanthus niruri, ikan nila

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Antioxidant and Hypoglycemic Properties of Seeds of Cucurbitaceae Family from Western Nepal

Journal of Health and Allied Sciences ◽

10.37107/jhas.118 ◽

2019 ◽

Vol 9 (2) ◽

pp. 1-5

Author(s):

Atisammodavardhana Kaundinnyayana ◽

Anita G.C ◽

Asmita Banstola ◽

Kishwor Poudel ◽

Nirmal Acharya ◽

...

Keyword(s):

Antioxidant Activity ◽

Radical Scavenging ◽

Ethanolic Extract ◽

Positive Control ◽

Lagenaria Siceraria ◽

Benincasa Hispida ◽

Ic50 Values ◽

In Vitro Antioxidant Activity

Introduction: Diabetes mellitus is an endocrine and metabolic disorder characterized by the defect in insulin secretion or insulin action or both. The association between diabetes mellitus and oxidative stress is well established. Cucurbitaceae is the largest family containing 120 genera mostly grown for its sweet and juicy fruit in warm climates all over the world. In the present work ethanolic extracts of the seeds of Trichosanthes cucumerina Linn (Chichinno), Lagenaria siceraria (Lauka), Cucurbita pepo (Pharsi), Luffa aegiptiaca (Ghiraula), and Benincasa hispida (Kubindo) collected from Western Nepal has been evaluated for their in vitro antioxidant activity and in vivo hypoglycemic effects. Methods: In vitro antioxidant activity was assessed by using DPPH free radical scavenging activities and their IC50 values were calculated. In vivo hypoglycemic effect was examined on normoglycemic rats. The clinical significance of ethanolic extract at the doses of 250 and 500 mg/kg body weight was investigated in 0, 30, 60, 120 and 180 minutes of oral administration. Metformin treated group was used as the positive control. Results: For DPPH radical scavenging action, the IC50 values of Trichosanthes cucumerina, Luffa aegiptiaca, Benincasa hispida, Cucurbita pepo, Lagenaria siceraria were found to be 60.72, 127.73, 49.63, 98.16 and 52.46 µg/ml respectively which were compared to the IC50 value of ascorbic acid (Positive control) which was found to be 38.11 µg/ml. The extracts of Benincasa hispida, and were having higher antioxidant activity and tested for in vivo hypoglycemic activity.In vivo administration of two doses of ethanolic extract of Lagenaria siceraria reduced the level of blood glucose while the best result was obtained at 250 mg/kg. Conclusions: Present study revealed promising antioxidant and hypoglycemic activity of ethanolic extract of Lagenaria siceraria. The further exploration of Lagenaria siceraria for its effective use in the traditional medicinal system is essential.

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The Role of BDNF on Aging-Modulation Markers

Brain Sciences ◽

10.3390/brainsci10050285 ◽

2020 ◽

Vol 10 (5) ◽

pp. 285

Author(s):

Claudio Molinari ◽

Vera Morsanuto ◽

Sara Ruga ◽

Felice Notte ◽

Mahitab Farghali ◽

...

Keyword(s):

Brain Aging ◽

Cerebral Atrophy ◽

Sirtuin 1 ◽

In Vivo Experiments ◽

Endogenous Production ◽

Age Related ◽

Protection Mechanisms ◽

Endogenous Protection

An important link between brain aging and a class of growth/survival factors called neurotrophins has recently been demonstrated. In particular, brain-derived neurotrophic factor (BDNF) plays a fundamental role during age-related synaptic loss, preventing cerebral atrophy and cognitive decline. The aim of the present study was to investigate whether the use of low dose BDNF sequentially kinetic activated (SKA) was able to counteract some mechanisms underlying the degeneration and aging of nervous tissue by increasing endogenous protection mechanisms. Both in vitro and in vivo experiments were performed to assess the ability of BDNF SKA to protect and regenerate survival-related molecular pathways, studying intestinal absorption in vitro and brain function in vivo. Our pioneering results show that BDNF SKA is able to induce the endogenous production of BDNF, using its receptor TrkB and influencing the apolipoprotein E expression. Moreover, BDNF SKA exerted effects on β-Amyloid and Sirtuin 1 proteins, confirming the hypothesis of a fine endogenous regulatory effect exerted by BDNF SKA in maintaining the health of both neurons and astrocytes. For this reason, a change in BDNF turnover is considered as a positive factor against brain aging.

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