Corrections for interferences and extraction conditions make a difference: use of the TBARS assay for lipid peroxidation of orthodoxSpartina pectinataand recalcitrantSpartina alternifloraseeds during desiccation

2011 ◽  
Vol 21 (2) ◽  
pp. 153-158 ◽  
Author(s):  
James H. Chappell ◽  
Marc Alan Cohn

AbstractLipid peroxidation and membrane damage are often proposed as causes of recalcitrant seed death, and the thiobarbituric acid reactive substances (TBARS) assay is commonly used to measure lipid peroxidation. However, several artefacts can cause an overestimation of TBARS values, and these have not been routinely addressed in experiments with recalcitrant seeds. In the present report, TBARS was assayed as recalcitrantSpartina alternifloraand orthodoxS. pectinataseeds were dried rapidly. Using the traditional Heath and Packer (1968) protocol with tissue extraction at 4°C,S. alterniflorahad higher overall TBARS values thanS. pectinata, and TBARS products increased when recalcitrantS. alternifloraand orthodoxS. pectinataseeds were dried. However, when corrections for interfering substances, such as sugars and anthocyanins, were made, the TBARS values between the two species were almost identical. When seeds were freeze-clamped in liquid nitrogen prior to extraction, TBARS did not increase during desiccation for either species. These findings may indicate that lipid peroxidation is not the cause of desiccation-induced death inS. alterniflora. Therefore, freeze-clamping during tissue extraction and corrections for TBARS interfering substances must be applied to avoid overestimation of lipid peroxidation values.

2011 ◽  
Vol 64 (7-8) ◽  
pp. 377-380 ◽  
Author(s):  
Aleksandra Novakov-Mikic ◽  
Snezana Brkic ◽  
Daniela Maric ◽  
Bojan Sekulic ◽  
Aleksandar Cetkovic ◽  
...  

Pre-eclampsia is characterized by increased lipid peroxidation and diminished antioxidant capacity. The aim of the study was to establish concentration of thiobarbituric acid reactive substances as a marker of lipid peroxidation in normal pregnancies and in pregnancies complicated with pre-eclampsia, and to estimate the possibility of using thiobarbituric acid reactive substances as a screening method for development of pre-eclampsia. The study was conducted at the Department of Obstetrics and Gynaecology, Clinical Centre of Vojvodina. The study included 57 singleton pregnancies, gestation >24 weeks, of which 29 were healthy pregnancies and 28 were with pre-eclampsia, defined as systolic arterial pressure of >90 mmHg, diastolic of >145 mmHg, and 24h proteinuria of >300mg. Thiobarbituric acid reactive substances concentrations evaluated by malondialdehyde equivalent standards (OxiSelect? TBARS Assay Kit (malondialdehyde Quantitation), Cell Biolabs? OxiSelect?) showed that oxidative stress was more evident in the group with pre-eclampsia, though not statistically significant (p= 0.107). There was no correlation of thiobarbituric acid reactive substance levels with gestation in either group. The differences between the level of thiobarbituric acid reactive substance concentrations in pre-eclampsia and healthy pregnancies indicate the possibility of using thiobarbituric acid reactive substances as a screening tool for the development of pre-eclampsia. Further studies with larger numbers of patients are needed in order to come to final conclusions.


2015 ◽  
Vol 3 (1) ◽  
pp. 41-47
Author(s):  
Nirjala Laxmi Madhikarmi ◽  
Kora Rudraiah Siddalinga Murthy

INTRODUCTION: The present study evaluated the modulatory effects of diphenylhydrazine induced experimental wistar albino rats and also to assess various biochemical parameters in whole blood and red blood cell lysate.MATERIALAND METHODS: Twenty male albino rats weighing 180-200 gm were selected for the study and divided in two groups; ten phenylhydrazine dihydrochloride (PHZ) induced anemia and ten healthy control. Thiobarbituric acid reactive substances and lipid hydroperoxide were measured as lipid peroxidation parameter. The antioxidant vitamins A, C and E and enzymatic antioxidants; catalase, glutathione peroxidase and superoxide dismutase were also assessed.RESULTS: Phenylhydrazine induced anemic rats showed a significant increase in the lipid peroxidation and decrease in the antioxidants as compared to healthy rats.CONCLUSION: The study concludes that phenylhydrazine induced experimental anemic albino rats showed increased oxidative stress than compared with healthy albino rats.Journal of Universal College of Medical Sciences Vol. 3, No. 1, 2015: 41-47 


2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Iulia Movileanu ◽  
Máryuri T. Núñez de González ◽  
Brian Hafley ◽  
Rhonda K. Miller ◽  
Jimmy T. Keeton

Fresh ground beef patties with (1) no antioxidant (control), (2) 0.02% butylated hydroxyanisole/butylated hydroxytoluene (BHA/BHT), (3) 3% dried plum puree, or (4) 0.25% rosemary extract were aerobically packaged, irradiated at target doses of 0, 1.5, or 2.0 kGy (1.7 and 2.3 kGy actual doses), and stored at C. The samples were evaluated for lipid oxidation on 0, 3, 7, 14, 21, and 28 days of storage after irradiation. When compared to the control, all antioxidant treatments were effective in retarding () irradiation-induced lipid oxidation during storage as determined by 2-thiobarbituric acid reactive substances (TBARs) values. Rosemary extracts had the same antioxidant effect () as BHA/BHT in irradiated and nonirradiated beef patties, followed by the dried plum puree treatment. Irradiation increased TBARs values, but no differences were noted in oxidation between irradiation dose levels.


2019 ◽  
Vol 11 (6) ◽  
pp. 401
Author(s):  
Patricia Wolkmer ◽  
Andressa M. G. Stumm ◽  
Luiz F. K. Borges ◽  
Eduarda P. T. Ferreira ◽  
Bruna Favaretto ◽  
...  

This experiment aims to evaluate the correlation between lipid peroxidation levels in serum and seminal plasma in equines. Also, it investigates the lipid peroxidation in extended semen samples and its effects and sperm motility during a 72 hr refrigeration period. Blood and semen were collected from fertile Crioulo stallions. Serum and seminal plasma lipid peroxidation levels were analyzed by thiobarbituric acid reactive substances (TBARS) immediately after semen collection. After addition of extender (hour = 0), diluted semen was refrigerated and stored at 5 °C. Semen analyses, TBARS and catalase activity were performed in extended semen at 0, 24, 48, and 72 hours. We noted that levels of plasma lipid peroxidation can be used as an indicative of seminal oxidative stress. Also, lipid peroxidation does not increase substantially during semen storage. Lipid peroxidation and the antioxidant enzyme catalase do not seem to be the major cause of loss and motility and consequently reduction in fertility in stallion semen during storage for 72 h at 5 °C.


1999 ◽  
Vol 87 (3) ◽  
pp. 1123-1131 ◽  
Author(s):  
G. Supinski ◽  
D. Nethery ◽  
D. Stofan ◽  
L. Szweda ◽  
A. DiMarco

The purpose of the present study was to determine whether it is possible to alter the development of fatigue and ablate free radical-mediated lipid peroxidation of the diaphragm during loaded breathing by administering oxypurinol, a xanthine oxidase inhibitor. We studied 1) room-air-breathing decerebrate, unanesthetized rats given either saline or oxypurinol (50 mg/kg) and loaded with a large inspiratory resistance until airway pressure had fallen by 50% and 2) unloaded saline- and oxypurinol-treated room-air-breathing control animals. Additional sets of studies were performed with animals breathing 100% oxygen. Animals were killed at the conclusion of loading, and diaphragmatic samples were obtained for determination of thiobarbituric acid-reactive substances and assessment of in vitro force generation. We found that loading of saline-treated animals resulted in significant diaphragmatic fatigue and thiobarbituric acid-reactive substances formation ( P < 0.01). Oxypurinol administration, however, failed to increase load trial time, reduce fatigue development, or prevent lipid peroxidation in either room-air-breathing or oxygen-breathing animals. These data suggest that xanthine oxidase-dependent pathways do not generate physiologically significant levels of free radicals during the type of inspiratory resistive loading examined in this study.


2006 ◽  
Vol 25 (2) ◽  
pp. 242-249 ◽  
Author(s):  
Freddy J. Troost ◽  
Robert-Jan M. Brummer ◽  
Guido R. M. M. Haenen ◽  
Aalt Bast ◽  
Rachel I. van Haaften ◽  
...  

Iron-induced oxidative stress in the small intestine may alter gene expression in the intestinal mucosa. The present study aimed to determine which genes are mediated by an iron-induced oxidative challenge in the human small intestine. Eight healthy volunteers [22 yr(SD2)] were tested on two separate occasions in a randomized crossover design. After duodenal tissue sampling by gastroduodenoscopy, a perfusion catheter was inserted orogastrically to perfuse a 40-cm segment of the proximal small intestine with saline and, subsequently, with either 80 or 400 mg of iron as ferrous gluconate. After the intestinal perfusion, a second duodenal tissue sample was obtained. Thiobarbituric acid-reactive substances, an indicator of lipid peroxidation, in intestinal fluid samples increased significantly and dose dependently at 30 min after the start of perfusion with 80 or 400 mg of iron, respectively ( P < 0.001). During the perfusion with 400 mg of iron, the increase in thiobarbituric acid-reactive substances was accompanied by a significant, momentary rise in trolox equivalent antioxidant capacity, an indicator of total antioxidant capacity ( P < 0.05). The expression of 89 gene reporters was significantly altered by both iron interventions. Functional mapping showed that both iron dosages mediated six distinct processes. Three of those processes involved G-protein receptor coupled pathways. The other processes were associated with cell cycle, complement activation, and calcium channels. Iron administration in the small intestine induced dose-dependent lipid peroxidation and a momentary antioxidant response in the lumen, mediated the expression of at least 89 individual gene reporters, and affected at least six biological processes.


Molecules ◽  
2020 ◽  
Vol 25 (8) ◽  
pp. 1765 ◽  
Author(s):  
Lorenzo Candela ◽  
Marialuisa Formato ◽  
Giuseppina Crescente ◽  
Simona Piccolella ◽  
Severina Pacifico

Marketed green teas (GTs) can highly vary in their chemical composition, due to different origins, processing methods, and a lack of standardization of GT-based products. Consequently, biological activities become difficult to correlate to the presence/content of certain constituents. Herein, ultra-high-performance liquid chromatography (UHPLC) combined with high-resolution tandem mass spectrometry (HR MS/MS) was successfully applied to six commercial GT products, extracted by ethanol sonication, to disclose their polyphenol profile beyond the well-known catechins. The relative abundance of each class of metabolites was correlated to antiradical and antilipoperoxidant data through hierarchical clustering analysis, since it reasonably affects the beneficial properties of the product that reaches the consumer. The thiobarbituric acid reactive substances (TBARS) assay demonstrated that GT extracts effectively counteracted the UV-induced lipoperoxidation of hemp oil, which is highly rich in Polyunsaturated Fatty Acids (PUFAs), and therefore highly unstable. The Relative Antioxidant Capacity Index (RACI) comprehensively emphasized that gunpower and blend in filter GTs appeared to be the less active matrices, and except for a GT-based supplement, the Sencha GT, which was particularly rich in flavonol glycosides, was the most active, followed by Bancha GT.


2011 ◽  
Vol 233-235 ◽  
pp. 2889-2892
Author(s):  
Xiao Ling Yu ◽  
Xue Bin Li ◽  
Hua Xiao ◽  
Han Jun Ma ◽  
Liang Cheng

Uniform design was adopted, and two factors, i.e., frozen at different time postmortem with six levels, thawing at different thawing rate with three levels, were considered. Thiobarbituric acid reactive substances (TBARS), pH and sensory evaluation were measured to study the different effects of the two factors considered. The results showed: All the two factors considered had significant effects on pH and TBARS, and the effects of the two factors on them were nonlinear. But they had no significant effects on sensory evaluation. TBARS values representing the storage quality of raw sausages was only affected by thawing rate in the test, and had the maximum at the thawing rate of 3cm/h thawed in 19.5°C still air. So when using frozen raw meat for sausage production, selecting an appropriate thawing condition is very necessary.


1994 ◽  
Vol 13 (12) ◽  
pp. 831-838 ◽  
Author(s):  
Hiroyuki Yokoyama ◽  
Toshiharu Horie ◽  
Shoji Awazu

1 Rat liver microsomal suspension containing NADPH and MgCl2 was incubated at 37°C with naproxen, a non-steroidal anti-inflammatory drug. Thiobarbituric acid reactive substances (TBA-RS), high molecular weight protein aggregates and fluorescent substances were formed in the microsomal suspension. 2 Chemiluminescence was produced from the microsomal suspension. This chemiluminescence production was well correlated to the TBA-RS formation, indicating that the chemiluminescence production was closely associated with the lipid peroxidation. 3 The addition of SKF-525A to the microsomal suspension inhibited the production of TBA-RS, chemiluminescence and 6-demethylnaproxen (6-DMN), the oxidative product of naproxen. Further, the antioxidant, α-tocopherol and singlet oxygen quenchers like histidine, dimethylfuran and 1,4-diazabicyclo[2,2,2]octane strikingly inhibited the productions of chemiluminescence and TBA-RS. 4 Neither naproxen nor 6-DMN caused lipid peroxidation in the absence of NADPH. Thus, lipid peroxidation and chemiluminescence during the oxidation of naproxen in liver microsomes was suggested to be provoked by reactive oxygen species and an origin of chemiluminescence was shown to be singlet oxygen.


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