scholarly journals Improvement of Flavonoids in Lemon Seeds on Oxidative Damage of Human Embryonic Kidney 293T Cells Induced by H2O2

2020 ◽  
Vol 2020 ◽  
pp. 1-10 ◽  
Author(s):  
Dingyi Yang ◽  
Yong Jiang ◽  
Yuqing Wang ◽  
Qianqian Lei ◽  
Xin Zhao ◽  
...  
Keyword(s):  
Survival Rate ◽  
Oxidative Damage ◽  
High Performance ◽  
Quantitative Polymerase Chain Reaction ◽  
Survival Rates ◽  
Human Embryonic Kidney ◽  
High Concentration ◽  
Stress Injury ◽  
Chromatography Analysis

In this study, flavonoids in lemon seeds (FLS) were used to assess its improvement on the oxidative damage of human embryonic kidney 293T cells (HEK 293T cells) induced by H2O2. In vitro experiments showed that the survival rates of HEK 293T cells treated with different flavonoid concentrations (50 μg/mL, 100 μg/mL, and 150 μg/mL) exceeded 95%, indicating no significant toxic effect. Compared with the normal group, H2O2 (0.3 mmol/L) resulted significantly in oxidative stress injury of HEK 293T cells. The survival rate of the damaged cells increased after treatment with flavonoids, and the survival rate of cells treated with a high concentration (150 μg/mL) of flavonoids was 76.2%. Flavonoids also effectively inhibited H2O2-induced apoptosis. At the same time, flavonoid treatment significantly reduced the malondialdehyde content in cells and increased the levels of catalase (CAT), superoxide dismutase (SOD), glutathione (GSH), and glutathione peroxidase (GSH-Px). Quantitative polymerase chain reaction (qPCR) and Western blot analysis also suggested that FLS upregulated mRNA and protein expressions of CAT, SOD (SOD1, SOD2), GSH (GSH1), and GSH-Px in H2O2-induced oxidative damage of HEK 293T cells. The high-performance liquid chromatography analysis demonstrated that FLS contained six compounds, including gallocatechin, caffeic acid, epicatechin, vitexin, quercetin, and hesperidin. FLS were proven to have a good antioxidant capacity in vitro and improve significantly the oxidative damage of HEK 293T cells induced by H2O2. The biological activity value warrants investigation in additional studies.

scholarly journals In Vitro Propagation, Huperzine A Content and Antioxidant Activity of Three Genotypic Huperzia serrata

Plants ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1112
Author(s):  
Yan Yang ◽  
Liangfang Dai ◽  
Decai Wu ◽  
Limin Dong ◽  
Yisheng Tu ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
High Performance ◽  
In Vitro Propagation ◽  
Huperzine A ◽  
Induction Medium ◽  
Naphthalene Acetic Acid ◽  
Regeneration Rate ◽  
Huperzia Serrata ◽  
Chromatography Analysis

Huperzia serrata is a traditional herb and endangered Chinese medicinal material, which has attracted much attention due to its production of Huperzine A (HupA). In vitro propagation of H. serrata is considered a new way to relieve the resource pressure of H. serrata. In this study, three different genotypic wild H. serrata were used for in vitro propagation. Then, the antioxidant activity and the content of HupA in the regenerated H. serrata were investigated. The results showed the survival rate of the explant was increased to 25.37% when using multiple sterilization processes. The best induction medium for H. serrata was the Schenk and Hildebrandt (SH) medium supplemented with 0.5 mg·L−1 Naphthalene acetic acid (NAA) and 0.1 mg·L−1 2,4-Dichlorophenoxyacetic acid (2,4-D), where the regeneration rate of the explant was to 57.04%. The best proliferation medium was the SH medium with NAA (1.0 mg·L−1), as the biomass of in vitro tissue increased 164.17 ± 0.41 times. High-performance liquid chromatography analysis showed that the in vitro culture of three genotypes could produce HupA and the content of HupA was 53.90–87.17 µg·g−1. The antioxidant experiment showed that the methanol extract of in vitro H. serrata had higher antioxidant activity than that of wild H. serrata. This study provides a reliable in vitro H. serrata culture protocol and laid an important foundation for the antioxidant capacity of the thallus and the content of HupA.

scholarly journals Abnormal physiological properties and altered cell wall composition in Streptococcus pneumoniae grown in the presence of clavulanic acid.

1997 ◽  
Vol 41 (3) ◽  
pp. 504-510 ◽  
Author(s):  
A Severin ◽  
E Severina ◽  
A Tomasz
Keyword(s):  
Streptococcus Pneumoniae ◽  
High Performance ◽  
Biochemical Properties ◽  
Beta Lactam ◽  
Chromatography Analysis ◽  
Peptide Composition ◽  
Increased Sensitivity ◽  
Altered Cell ◽  
Quantitative Changes

Subinhibitory concentrations of clavulanate caused premature induction of stationary-phase autolysis, sensitization to lysozyme, and reductions in the MICs of deoxycholate and penicillin for Streptococcus pneumoniae. In the range of clavulanate concentrations producing these effects, this beta-lactam compound was selectively bound to PBP 3. Cell walls isolated from pneumococci grown in the presence of clavulanate showed increased sensitivity to the hydrolytic action of purified pneumococcal autolysin in vitro. High-performance liquid chromatography analysis of the peptidoglycan isolated from the clavulanate-grown cells showed major qualitative and quantitative changes in stem peptide composition, the most striking feature of which was the accumulation of peptide species carrying intact D-alanyl-D-alanine residues at the carboxy termini. The altered biological and biochemical properties of the clavulanate-grown pneumococci appear to be the consequences of suppressed D,D-carboxypeptidase activity.

scholarly journals CHARACTERIZATION OF MOLLUSCICIDAL COMPONENT OF Moringa oleifera LEAF AND Momordica charantia FRUITS AND THEIR MODES OF ACTION IN SNAIL Lymnaea acuminata

2013 ◽  
Vol 55 (4) ◽  
pp. 251-259 ◽  
Author(s):  
Aparna Upadhyay ◽  
Vinay K. Singh ◽  
Dinesh K. Singh
Keyword(s):  
High Performance ◽  
Moringa Oleifera ◽  
Momordica Charantia ◽  
Chromatography Analysis ◽  
Leaf Powder ◽  
Fruit Powder ◽  
Lymnaea Acuminata ◽  
Molluscicidal Activity

SUMMARY The molluscicidal activity of the leaf powder of Moringa oleifera and lyophilized fruit powder of Momordica charantia against the snail Lymnaea acuminata was time and concentration dependent. M. oleifera leaf powder (96 h LC50: 197.59 ppm) was more toxic than M. charantia lyophilized fruit powder (96 h LC50: 318.29 ppm). The ethanolic extracts of M. oleifera leaf powder and Momordica charantia lyophilized fruit powder were more toxic than other organic solvent extracts. The 96 h LC50 of the column purified fraction of M. oleifera leaf powder was 22.52 ppm, while that of M. charantia lyophilized fruit powder was 6.21 ppm. Column, thin layer and high performance liquid chromatography analysis show that the active molluscicidal components in M. oleifera leaf powder and lyophilized fruit of M. charantia are benzylamine (96 h LC50: 2.3 ppm) and momordicine (96 h LC50: 1.2 ppm), respectively. Benzylamine and momordicine significantly inhibited, in vivo and in vitro, the acetylcholinesterase (AChE), acid and alkaline phosphatase (ACP/ALP) activities in the nervous tissues of L. acuminata. Inhibition of AChE, ACP and ALP activity in the nervous tissues of L. acuminata by benzylamine and momordicine may be responsible for the molluscicidal activity of M. oleifera and M. charantia fruits, respectively.

83 VITRIFICATION OF IMMATURE AND IN VITRO-MATURED HORSE OOCYTES

2006 ◽  
Vol 18 (2) ◽  
pp. 149 ◽  
Author(s):  
L. Bogliolo ◽  
F. Ariu ◽  
I. Rosati ◽  
M. T. Zedda ◽  
S. Pau ◽  
...  
Keyword(s):  
Survival Rate ◽  
Ethylene Glycol ◽  
Survival Rates ◽  
Germinal Vesicle ◽  
Cumulus Cells ◽  
Hoechst 33342 ◽  
Nuclear Maturation ◽  
And Control ◽  
Cryopreservation Protocol

Few attempts have been carried out to cryopreserve equine oocytes, and an effective cryopreservation protocol is not defined yet. Studies were conducted to compare the viability of immature and in vitro-matured horse oocytes vitrified by the minimal volume cooling (MVC) cryotop vitrification method (Kuwayama et al. 2005 Reprod. BioMed. Online 11, 300–308). Oocytes were recovered from slaughterhouse ovaries and divided, on the basis of the morphology of cumulus cells, into cumulus-expanded (CE) and cumulus-compacted (CC) oocytes. Groups of CC and CE oocytes were vitrified immediately after recovery [germinal vesicle (GV) stage] or matured in vitro (IVM) and cryopreserved at the MII stage as follows: oocytes were incubated 30 min in TCM-199 + 20% FCS + 10% ethylene glycol (EG) + 10% DMSO, followed by 20 min in TCM-199 + 20% FCS + 20% EG + 20% DMSO + 0.25 M sucrose, loaded in cryotops (2 µL), and plunged into liquid nitrogen. Warming was performed at 38.5°C by washing the oocytes in TCM-199 + 20% FCS with decreasing sucrose concentrations (1.25 M, 0.62 M, 0.31 M). After warming oocytes cryopreserved at the GV stage were matured in vitro for 24 h (CE) or 36 h (CC) in TCM-199 + 10% FCS + FSH, LH each at (0.1 UI/mL) + cysteamine, fixed, and stained with glycerol-Hoechst 33342 to assess nuclear maturation. Oocytes vitrified at the MII stage were in vitro cultured for 2 h to evaluate their morphological survival on the basis of the presence of an intact zona pellucida and membrane. Nonvitrified oocytes undergoing the same maturation protocol were used as controls. Results (Table 1) indicated that the survival rate of oocytes vitrified at the GV stage, after IVM, was similar between CE and CC oocytes (43.6% vs 42.6%). Significantly (P < 0.01) higher numbers of vitrified CE MII oocytes (52.9%) survived, compared to CC (34.8%), after 2-h culture. The percentages of viable MII oocytes from CE and CC GV vitrified oocytes were 43.6% and 40.9% respectively and were comparable to those from vitrified MII oocytes (CE, 52.9%; CC, 34.8%) and control oocytes (CE, 56.4%; CC, 53.3%). In conclusion, the results of this study showed that vitrification by the MCV Cryotop method of horse oocytes at either the GV or the MII stage allows a similar number of viable mature oocytes to be recovered. Table 1. Maturation and survival rates of immature and mature equine oocytes vitrified by the MCV Cryotop method

scholarly journals Effect of Drying Process, Encapsulation, and Storage on the Survival Rates and Gastrointestinal Resistance of L. salivarius spp. salivarius Included into a Fruit Matrix

2020 ◽  
Vol 8 (5) ◽  
pp. 654
Author(s):  
Ester Betoret ◽  
Noelia Betoret ◽  
Laura Calabuig-Jiménez ◽  
Cristina Barrera ◽  
Marco Dalla Rosa
Keyword(s):  
Survival Rate ◽  
Physicochemical Properties ◽  
Freeze Drying ◽  
Molecular Mechanisms ◽  
Survival Rates ◽  
In Vitro Digestion ◽  
Hot Air ◽  
Freeze Dried ◽  
And Storage

In a new probiotic food, besides adequate physicochemical properties, it is necessary to ensure a minimum probiotic content after processing, storage, and throughout gastrointestinal (GI) digestion. The aim of this work was to study the effect of hot air drying/freeze drying processes, encapsulation, and storage on the probiotic survival and in vitro digestion resistance of Lactobacillus salivarius spp. salivarius included into an apple matrix. The physicochemical properties of the food products developed were also evaluated. Although freeze drying processing provided samples with better texture and color, the probiotic content and its resistance to gastrointestinal digestion and storage were higher in hot air dried samples. Non-encapsulated microorganisms in hot air dried apples showed a 79.7% of survival rate versus 40% of the other samples after 28 days of storage. The resistance of encapsulated microorganisms to in vitro digestion was significantly higher (p ≤ 0.05) in hot air dried samples, showing survival rates of 50–89% at the last stage of digestion depending on storage time. In freeze dried samples, encapsulated microorganisms showed a survival rate of 16–47% at the end of digestion. The different characteristics of the food matrix after both processes had a significant effect on the probiotic survival after the GI digestion. Documented physiological and molecular mechanisms involved in the stress response of probiotic cells would explain these results.

DEVELOPMENT OF AN INTRODUCTION PROTOCOL FOR STRAWBERRY PLANTS INTO IN VITRO CULTURE

2020 ◽  
Vol 5 (86) ◽  
pp. 45-50
Author(s):  
O.V. Mazneva ◽  
◽  
L.V. Tashmatova ◽  
T.M. Khromova ◽  
V.V. Shakhov ◽  
...  
Keyword(s):  
Survival Rate ◽  
In Vitro Culture ◽  
Plant Material ◽  
Survival Rates ◽  
Strawberry Plant ◽  
Active Growth ◽  
High Viability ◽  
Average Survival ◽  
Average Survival Rate

The research was conducted in order to develop an effective protocol for introducing strawberry plants into in vitro culture. The objects of the research were the most popular varieties of strawberries of domestic and foreign selection: Tsaritsa, Bereginya, Florence, Frida, Kimberly, etc. Mercurial preparations mertiolate at a concentration of 0.01% and sulema at a concentration of 0.1% were used as sterilizing agents. The isolation of explants was performed in several periods: the beginning of the growth was in February, active growth was in June, the decline of growth was in August. The studies have shown that the maximum aseptic cultures were obtained when processing strawberry plant material with mercurycontaining sulema preparation in the concentration of 0.1%. At the first stage of micropropagation, explants had a high viability during all periods of the isolation, the average survival rate for varieties was 74.8-80.7%. A significant influence of the genotype (varietal characteristics) on the survival rates of explants was noted. The number of explants suitable for cloning did not depend on the overall level of regeneration. Stabilization of the crop during winter introduction was much faster than in other periods. Using the winter term of the isolation of strawberry explants allowed to increase the yield of explants capable of further cloning, accelerate the stabilization of the culture in vitro and reduce the time for obtaining micro-plants suitable for planting in non-sterile conditions. On average, 75.2% of explants capable of further cloning for the varieties were obtained. As a result of the research, the conditions and methods for obtaining the largest number of viable sterile strawberry explants were worked out, which will be included into the process of reproduction in vitro and further research.

scholarly journals Improved development by Taxol pretreatment after vitrification of in vitro matured porcine oocytes

Reproduction ◽  
2006 ◽  
Vol 131 (4) ◽  
pp. 795-804 ◽  
Author(s):  
Wen-Qing Shi ◽  
Shi-En Zhu ◽  
Dong Zhang ◽  
Wei-Hua Wang ◽  
Guo-Liang Tang ◽  
...  
Keyword(s):  
Survival Rate ◽  
Embryo Development ◽  
Survival Rates ◽  
Developmental Competence ◽  
Fluorescein Diacetate ◽  
Microtubule Organization ◽  
Cell Stage ◽  
Parthenogenetic Activation ◽  
Parthenogenetic Development

This study was designed to examine the effect of Taxol pretreatment on vitrification of porcine oocytes matured in vitro by an open pulled straw (OPS) method. In the first experiment, the effect of Taxol pretreatment and fluorescein diacetate (FDA) staining on parthenogenetic development of oocytes was evaluated. In the second experiment, viability, microtubule organization and embryo development of oocytes were assessed after oocytes were exposed to vitrification/warming solutions or after vitrification with or without Taxol pretreatment. The results showed that Taxol pretreatment and/or FDA staining did not negatively influence the oocyte’s developmental competence after parthenogenetic activation. After being exposed to vitrification/warming solutions, the survival rate (83.3%) of the oocytes was significantly (P < 0.05) reduced as compared with that in the control (100%). Vitrification/warming procedures further reduced the survival rates of oocytes regardless of oocytes being treated with (62.1%) or without (53.8%) Taxol. The proportions of oocytes with normal spindle configuration were significantly reduced after the oocytes were exposed to vitrification/warming solutions (38.5%) or after vitrification with (10.3%) or without (4.1%) Taxol pretreatment as compared with that in control (76.8%). The rates of two-cell-stage (5.6–53.2%) embryos at 48 h and blastocysts (0–3.8%) at 144 h after activation were significantly reduced after exposure to vitrification/warming solutions or after vitrification as compared with control (90.9% and 26.6% respectively). However, the proportion of vitrified oocytes developed to two-cell stage was significantly higher when oocytes were pretreated with (24.3%) than without (5.6%) Taxol. These results indicate that pretreatment of oocytes with Taxol before vitrification helps to reduce the damage induced by vitrification and is a potential way to improve the development of vitrified porcine oocytes.

Physiological concentrations of retinoic acid suppress the osteoblastic differentiation of fetal rat calvaria cells in vitro

1995 ◽  
Vol 133 (3) ◽  
pp. 335-341 ◽  
Author(s):  
Keiji Ohishi ◽  
Seiji Nishikawa ◽  
Toshihiko Nagata ◽  
Noriyuki Yamauchi ◽  
Hiroyuki Shinohara ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
High Performance ◽  
Osteoblastic Differentiation ◽  
Dependent Manner ◽  
High Concentration ◽  
Physiological Concentration ◽  
Rat Calvaria ◽  
Fetal Rat ◽  
School Of Dentistry

Ohishi K, Nishikawa S, Nagata T, Yamauchi N, Shinohara H, Kido J, Ishida H. Physiological concentrations of retinoic acid suppress the osteoblastic differentiation of fetal rat calvaria cells in vitro. Eur J Endocrinol 1995;133:335–41. ISSN 0804–4643 The effects of retinoic acid (RA) on osteoblastic differentiation and activity were studied in fetal rat calvaria cells cultured for up to 24 days. Fetal bovine serum used for the experiments was treated with an anion-exchange resin to remove endogenous RA. The depletion of RA in the treated serum was confirmed by high-performance liquid chromatography and tritiated RA tracing. Under the culture conditions employed, the continuous presence of RA for 14 days at 10−9 mol/l or higher decreased both alkaline phosphatase (ALP) activity on day 12 and the number of bone nodules on day 14 in a dose-dependent manner. Short-term (24 h) exposure to RA at 10−8 mol/l, which is a physiological concentration, decreased and increased the levels of ALP and osteopontin mRNA on day 6, respectively. Retinoic acid at 10−8 mol/l also increased the level of osteocalcin mRNA on day 12. However, these effects were not obvious at later stages (days 18 and 24). At a high concentration (10−6 mol/l). RA increased the level of osteopontin mRNA on day 6 and decreased the levels of ALP and osteocalcin mRNA irrespective of culture period. These results suggest that, at physiological concentrations, RA suppresses the differentiation of osteoprogenitor cells and regulates osteoblastic functions. H Ishida, Department of Periodontology and Endodontology. Tokushima University School of Dentistry, 3-18-15 Kuramoto-cho, Tokushima 770, Japan

scholarly journals Novel Type of Heme-Dependent Oxygenase Catalyzes Oxidative Cleavage of Rubber (Poly-cis-1,4-Isoprene)

2004 ◽  
Vol 70 (12) ◽  
pp. 7388-7395 ◽  
Author(s):  
Reinhard Braaz ◽  
Peter Fischer ◽  
Dieter Jendrossek
Keyword(s):  
Natural Rubber ◽  
High Performance ◽  
Natural Rubber Latex ◽  
Culture Fluid ◽  
Oxidative Cleavage ◽  
Covalent Attachment ◽  
Rubber Latex ◽  
Chromatography Analysis ◽  
Cloned Gene

ABSTRACT An extracellular protein with strong absorption at 406 nm was purified from cell-free culture fluid of latex-grown Xanthomonas sp. strain 35Y. This protein was identical to the gene product of a recently characterized gene cloned from Xanthomonas sp., as revealed by determination of m/z values and sequencing of selected isolated peptides obtained after trypsin fingerprint analysis. The purified protein degraded both natural rubber latex and chemosynthetic poly(cis-1,4-isoprene) in vitro by oxidative cleavage of the double bonds of poly(cis-1,4-isoprene). 12-Oxo-4,8-dimethyltrideca-4,8-diene-1-al (m/z 236) was identified and unequivocally characterized as the major cleavage product, and there was a homologous series of minor metabolites that differed from the major degradation product only in the number of repetitive isoprene units between terminal functions, CHO-CH2— and —CH2-COCH3. An in vitro enzyme assay for oxidative rubber degradation was developed based on high-performance liquid chromatography analysis and spectroscopic detection of product carbonyl functions after derivatization with dinitrophenylhydrazone. Enzymatic cleavage of rubber by the purified protein was strictly dependent on the presence of oxygen; it did not require addition of any soluble cofactors or metal ions and was optimal around pH 7.0 at 40°C. Carbon monoxide and cyanide inhibited the reaction; addition of catalase had no effect, and peroxidase activity could not be detected. The purified protein was specific for natural rubber latex and chemosynthetic poly(cis-1,4-isoprene). Analysis of the amino acid sequence deduced from the cloned gene (roxA [rubber oxygenase]) revealed the presence of two heme-binding motifs (CXXCH) for covalent attachment of heme to the protein. Spectroscopic analysis confirmed the presence of heme, and approximately 2 mol of heme per mol of RoxA was found.

scholarly journals Effect of Different Isorhamnetin of Different Density Cultured Rat C6 Glioma Cells in Vitro

2018 ◽  
Vol 2 (1) ◽  
Author(s):  
Jinhua Yang ◽  
Zhuanni Sun
Keyword(s):  
Survival Rate ◽  
Brain Tissue ◽  
High Performance ◽  
Glioma Cells ◽  
C6 Glioma ◽  
C6 Glioma Cells ◽  
Control Group ◽  
Brain Glioma ◽  
Apoptosis And Necrosis

Objective To explore the influence of different concentrations of isorhamnetin on C6 glioma cell morphology. Methods Set the blank control group, blank solvent control group and reagent group of four concentration, the growth of cells were observed under microscope; MTT assay was used to test the effect of isorhamnetin on cultured C6 glioma cells, as well as calculate the cell inhibition rate and survival rate; flow cytometry was used to check the detection peak and detection rate of Isorhamnetin group and negative control apoptosis group, and analyzed the relationship between different concentrations of isorhamnetin and C6 glioma cell apoptosis rate; total protein was extracted from cells, and used Western blotting to detected total AKT protein and Ser473 AKT protein loci in cells; used SD rats to construct brain glioma model, feed isorhamnetin plain to them for five days, and then used HPLC to detect plasma, liver, brain tissue content. Results Under the observation of inverted microscope and image analysis, after using Isorhamnetin, tumor cells appear apoptosis and necrosis change. Display with different Isorhamnetin MTT colorimetric method shows that the higher the concentration of added Isorhamnetin, the worse the growth rate of C6 glioma cells in vitro, and the higher the Inhibitory rate, the lower survival rate. The flow cytometric detection shows the C6 glioma cells which is added 40 ug/ul Isorhamnetin have the highest rate of apoptosis. After adding 80 μ g/ μ l concentration of the isorhamnetin, C6 glioma cells have the lowest survival rate. Western blot test shows the AKT protein and Ser473 total site AKT protein density is in reverse proportion to the increase of the concentration of the isorhamnetin. High performance liquid chromatographic method has determined that there are isorhamnetin in both the rat plasma and brain tissue, which shows that the plasma and tissue all have different isorhamnetin  distribution,  and  isorhamnetin mainly exist in the brain tissue. Conclusion Low concentration  of  isorhamnetin  can  induce apoptosis  of  C6  glioma  cells,  and  high concentration of isorhamnetin can lead to apoptosis and necrosis of C6 glioma cells in vitro, which has obvious inhibitory effect on the growth of glioma cells, and the mechanism is closely related to PI3K/AKT pathway, and in SD rat brain glioma model , the high performance liquid chromatography was used to detect the content of plasma and brain tissue, which indicated the isorhamnetin has target in brain tissue, which provided  experimental  evidence  for  the development and utilization of isorhamnetin in mice.

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