Enzyme Protein Content Research Articles

Physiological effects of short-term copper stress on rape (Brassica napus L.) seedlings and the alleviation of copper stress by attapulgite clay in growth medium

Rape (Brassica napus L.) seedlings grown in vermiculite-based medium were subjected to short-term copper stress. With the increase of exogenous CuCl2 concentration, photosystem II (PSII) photochemistry and the chlorophyll (Chl) content of rape seedling leaves decreased, while Cu (copper) content of the seedlings, the levels of the soluble protein content, reactive oxygen species (ROS) production, and activities of antioxidant enzymes of the seedling leaves increased. Under the condition without copper stress, there was no significant differences in the measured physiological indexes between the seedlings grown in vermiculite and those grown in the attapulgite clay (AC)/vermiculite mixture with the volume ratio at 1:80, 1:50, or 1:30. After subjected to short-term copper stress, the rape seedlings grown in the mixture of AC/vermiculite had higher levels of Chl content and PSII photochemistry but lower levels of the Cu content, soluble protein content, ROS production, and activities of antioxidant enzymes, compared with the leaves of the seedlings grown in vermiculite. The volume ratio of AC/vermiculite at 1:50 or 1:30 seemed to be the most optimized content in alleviating the physiological effects by the short-term copper stress. These results indicate that AC at moderate content in the growth medium can alleviate the physiological stress of the rape seedlings when excess Cu emerges in the growth medium.

Effect of cyanide by sodium nitroprusside (SNP) application on germination, antioxidative system and lipid peroxidation of Senna macranthera seeds under saline stress

Abstract: The effects of NO donors on germination under saline stress have been much investigated for many species, however, there are reports that the effect caused by donors are effects of cyanide present. The aim of this study was to evaluate the effects of sodium nitroprusside (SNP) on germination, antioxidative system and lipid peroxidation of Senna macranthera seeds under saline stress. The osmotic potentials of -0.4 and -0.5 MPa of NaCl were used, as well as the concentration of 100 μM of sodium nitroprusside, inactive sodium nitroprusside. Germination rate, imbibition curves, antioxidant enzyme activity, lipid peroxidation, and protein content were evaluated. Similar effects were observed for sodium nitroprusside and inactive sodium nitroprusside, indicating that these effects were related to the release of the cyanide present in sodium nitroprusside. The sodium nitroprusside (SNP) improves the germination of Senna macranthera seeds under salt stress conditions, through release of cyanide. The cyanide favored germination, reducing lipid peroxidation and increasing the activity of the antioxidant enzymes.

Biomarker Responses to Polycyclic Aromatic Hydrocarbons in the Native Fish Ramnogaster arcuata, South America

Quantification of polycyclic aromatic hydrocarbons (PAHs) in Bahia Blanca Estuary (BBE, Argentina) fish samples (Ramnogaster arcuata) was performed to evaluate the environmental impact through anthropogenic activity. In addition, several metabolic enzyme activities (Aspartate aminotransferase—AST, Alanine aminotransferase—ALT, Lactate dehydrogenase—LDH, Creatine kinase—CK and, Alkaline phosphatase—ALP), protein content and lipid peroxidation as oxidative stress biomarker were analyzed in muscle and liver and related to tisular PAHs levels. Results showed low to moderate PAHs levels in R. arcuata muscle (9.34–41.25 ng/g, wet weight) with a marked predominance of two/three ringed compounds (phenanthrene > naphthalene > acenaphthene > acenaphthylene > fluoranthene). Fluoranthene, pyrene, benzo-[b]fluoranthene and benzo-[a]pyrene concentrations correlated positively with hepatic AST and ALT and negatively with muscular proteins and hepatic lipid peroxidation. 2-metil-naphthalene and acenaphthene levels correlated negatively with LDH in muscle and positively with lipid peroxidation in liver tissue. Correlation of PAHs with metabolic enzymes, proteins and lipid peroxidation indicated a differential metabolization and suggesting that hepatic AST/ALT could be used as PAHs biotransformation biomarkers and muscular LDH as a stress oxidation biomarker in R. arcuata. In addition, CK activity was suggested as a good index of muscular health. The obtained results highlighted the significance of using a set of integrated biomarkers to assess PAHs toxicity in fish inhabiting their natural ambient and confirm that R. arcuata could be used as a good bioindicator for marine areas.

Manganese oxide nanoparticle-induced changes in growth, redox reactions and elicitation of antioxidant metabolites in deadly nightshade (Atropa belladonna L.)

Nanoparticles (NPs) can stimulate biosynthesis of bioactive compounds in plants. Mn NP is a less explored NP, and very little is known about interactions between Mn2O3 NPs and plants. The present study was conducted to explore the potential effects of Mn2O3 NPs on morphology, physiology and secondary metabolism of shoot tip cultures in the medicinal plant deadly nightshade (Atropa belladonna). The MS culture medium was augmented with different concentrations (25, 50, 100 and 200 mg L−1) of Mn2O3 NPs, and the medium with or without manganese sulfate (MnSO4·4H2O) was considered as positive and negative control, respectively. The addition of Mn2O3 NPs to culture media led to changes in morphology (root and shoot number per explants, root and shoot length, and root and shoot fresh weight) and physiology (leaf relative water content, total chlorophyll, hydrogen peroxide and malondialdehyde contents, electrolyte leakage and membrane stability index) by altering the protein content and activation of antioxidant and defense enzymes (superoxide dismutase, peroxidase, catalase and ascorbate peroxidase) and regulation of pharmacologically active metabolites (total phenolics, flavonoids and alkaloids) biosynthesis in a dose-dependent manner. All the extracts obtained from plantlets grown in various culture media showed DPPH free radical scavenging and Fe2+- chelating activities to different degrees. The index of integrated biological marker response (IBR/n) for different antioxidant enzymes was differentially influenced by various concentrations of Mn2O3 NPs. An UPGMA dendrogram of cluster analysis was also constructed, which divided all the studied treatments into three major groups (toxic, positive and inconsequential). At appropriate dosage (25 mg L−1), Mn2O3 NPs were found to be stimulant of both plant growth and metabolic processes such as production of alkaloids. Thus, Mn2O3 NPs would be a novel extracellular elicitor of secondary metabolites for tissue culture systems in the agricultural biotechnology industry.

Insecticidal Activity of Moringa oleifera Synthesized Silver and Zinc Nanoparticles against the House Fly, Musca domestica L.

Green synthesizednanoparticles have been studied as newand more effective biopesticides against insects of medical importance and thisfor their eco-friendly aspects. In the current study Moringa oleiferaleaf extract (Mo-LE) was used to synthesis silver nanoparticles (Mo-AgNPs) andzinc oxide nanoparticles (Mo-ZnONPs). Low concentrations of Mo-LE, Mo-AgNPs andMo-ZnONPs showed the larvicidal and pupicidal toxicityagainst Musca domestica. In larvicidal bioassay, the values of LC50were 16.50, 2.03 and 6.41 mg/ml for Mo-LE, Mo-AgNPs andMo-ZnONPs,respectively. In addition, the LC50 was determined against pupae,which were129.77, 9.604 and 17.10 mg/ml for Mo-LE, Mo-AgNPs andMo-ZnONPs,respectively. Moreover, treatment larvae with LC50's of Mo-LE, Mo-AgNPs andMo-ZnONPsproduced a significant reduction (P<0.05) in the fecundity of femaleand a highly significant reduction (P<0.01) in the egg hatchability.The total protein content and activities of nonspecific esterases, acetylcholine esterase, and glutathione S-transferase enzymes were significantly declined afterlarvae had fed on LC50 Mo-AgNPs andMo-ZnONPstreated diets as compared with the control. This study indicates that it isfeasible to use M. oleifera synthesized nanoparticles as effective candidates to develop newer and cheapcontrol tools for immature stages of M. domestica.

Oxygen conserving mitochondrial adaptations in the skeletal muscles of breath hold divers.

BackgroundThe performance of elite breath hold divers (BHD) includes static breath hold for more than 11 minutes, swimming as far as 300 m, or going below 250 m in depth, all on a single breath of air. Diving mammals are adapted to sustain oxidative metabolism in hypoxic conditions through several metabolic adaptations, including improved capacity for oxygen transport and mitochondrial oxidative phosphorylation in skeletal muscle. It was hypothesized that similar adaptations characterized human BHD. Hence, the purpose of this study was to examine the capacity for oxidative metabolism in skeletal muscle of BHD compared to matched controls.MethodsBiopsies were obtained from the lateral vastus of the femoral muscle from 8 Danish BHD and 8 non-diving controls (Judo athletes) matched for morphometry and whole body VO2max. High resolution respirometry was used to determine mitochondrial respiratory capacity and leak respiration with simultaneous measurement of mitochondrial H2O2 emission. Maximal citrate synthase (CS) and 3-hydroxyacyl CoA dehydrogenase (HAD) activity were measured in muscle tissue homogenates. Western Blotting was used to determine protein contents of respiratory complex I-V subunits and myoglobin in muscle tissue lysates.ResultsMuscle biopsies of BHD revealed lower mitochondrial leak respiration and electron transfer system (ETS) capacity and higher H2O2 emission during leak respiration than controls, with no differences in enzyme activities (CS and HAD) or protein content of mitochondrial complex subunits myoglobin, myosin heavy chain isoforms, markers of glucose metabolism and antioxidant enzymes.ConclusionWe demonstrated for the first time in humans, that the skeletal muscles of BHD are characterized by lower mitochondrial oxygen consumption both during low leak and high (ETS) respiration than matched controls. This supports previous observations of diving mammals demonstrating a lower aerobic mitochondrial capacity of the skeletal muscles as an oxygen conserving adaptation during prolonged dives.

Antioxidant Responses of Ragweed Leaf Beetle Ophraella communa (Coleoptera: Chrysomelidae) Exposed to Thermal Stress.

Ophraella communa LeSage is an effective biological control agent of common ragweed, Ambrosia artemisiifolia L., which competes with crops and causes allergic rhinitis and asthma. However, thermal stress negatively affects the developmental fitness and body size of this beetle. High temperatures cause a variety of physiological stress responses in insects, which can cause oxidative damage. We investigated the total protein content and activity of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), and peroxidases (PODs) in O. communa adults when its different developmental stages were exposed to high temperatures (40, 42, and 44°C) for 3 h each day for 3, 5, 5, and 5 days, respectively (by stage), and a whole generation to high temperatures (40, 42, and 44°C) for 3 h each day. A control group was reared at 28 ± 2°C. Under short-term daily phasic high-temperature stress, total protein contents were close to the control as a whole; overall, SOD activities increased significantly, CAT activities were closer to or even higher than the control, POD activities increased at 40°C, decreased at 42 or 44°C; stage-specific response was also observed. Under long-term daily phasic high-temperature stress, total protein content increased significantly at 44°C, SOD activities increased at higher temperatures, decreased at 44°C; CAT activities of females increased at ≤42°C, and decreased at 44°C, CAT activities of males decreased significantly; POD activities of females increased at 40°C, decreased at ≥42°C, POD activities of males decreased at 44°C; and antioxidant enzymes activities in females were significantly higher than those in males. Antioxidative enzymes protect O. communa from oxidative damage caused by thermal stress.

Effect of 24-Epibrassinolide on Antioxidative Defence System Against Lead-Induced Oxidative Stress in The Roots of Brassica juncea L. Seedlings

Lead (Pb) toxicity causes oxidative stress by increasing the production of reactive oxygen species. The aim of the present study was to investigate the role of 24-epibrassinolide (24-EBL) on the antioxidant defence system as a response to Pb stress in Brassica juncea L. Surface-sterilized seeds were exposed to Pb ion (0 and 2 mM) toxicity in Petri dishes and subsequently, the seeds were sprayed with either (i) deionized water or (ii) different concentrations (10–12, 10–10, and 10–8 M) of 24-EBL on alternate days. After nine days, the roots of the B. juncea seedlings were harvested to analyze the heavy metal content, root length, hydrogen peroxide level, lipid peroxidation, total protein content and activities of the antioxidant enzymes (superoxide dismutase, catalase, ascorbate peroxidase, peroxidase, glutathione reductase and glutathione-S-transferase). According to our results, the Pb ions accumulated by the B. juncea roots led to oxidative stress by increasing the level of H2O2 and malondialdehyde, and thus, increased the activity of the antioxidative enzymes (except for catalase) and the growth and total protein content decreased. Whereas, the 24-EBL treatment to the roots of Pb stressed seedlings was able to alleviate the Pb-induced oxidative stress. Upon the application of 24-EBL, a reduction in Pb accumulation, H2O2 and malondialdehyde levels as well as an increased total protein content and activity of antioxidative enzymes detoxifying hydrogen peroxide (catalase, ascorbate peroxidase and peroxidase) were observed. As a result, the stress protective properties of 24-EBL depending on concentration in B. juncea roots were revealed in this study.

Bisphenol A and its analogs bisphenol B, bisphenol F, and bisphenol S: Comparative in vitro and in vivo studies on the sperms and testicular tissues of rats

Bisphenol A (BPA) is used as the main component of many consumer products such as infant's feeding bottles, coatings of beverages, and food cans. BPA can migrate into the environment, and it has been detected in the saliva, blood, and food. BPA leakage from many consumer products resulted in a ban on its use in many countries where alternatives to BPA were introduced into the market. BPA alternatives such as bisphenol B (BPB), bisphenol F (BPF), and bisphenol S (BPS) have a similar chemical structure and binding ability for estrogen receptor (ER), which shows toxicological effects in animals. In the present study, comparative effects of exposure to BPA and its analogs BPB, BPF, and BPS on testosterone concentration in the rat testis were evaluated by in vitro and in vivo approaches in which oxidative stress markers and antioxidant enzyme activities in reproductive tissues were determined. In the in vivo study, male rats were exposed to different concentrations of BPA and its analogs BPB, BPF, and BPS (5, 25, and 50 mg/kg/day) for 28 days. In the in vitro exposure study, antioxidant enzyme activities and oxidative stress markers were induced in the testes, whereas testosterone production was reduced. In the in vivo exposure study, we observed that antioxidant enzyme activities and protein content were reduced, whereas reactive oxygen species and lipid profile were increased in the treated groups compared to the control group. The present comparative study on BPA and its analogs, namely, BPB, BPF, and BPS suggests the toxic effect of these chemicals on the testes and spermatogenesis, and we also observed that these chemicals induce oxidative stress in the reproductive tissues of male rats.

Removing of Benzo[a]pyrene using the Isolated Fungi from Petroleum-polluted Soils

Benzo[a]pyrene, belonging to polycyclic aromatic hydrocarbons, is one of the most important industrial pollutants. This research was aimed to evaluate some fungal strains, with petroleum removing potency, for degradation of BaP from BaP-polluted media and also evaluation of Enzyme activity and protein content in the fungi growing in BaP-polluted media. In a field study seven fungal specie were isolated from Tehran oil refinery and cultured in potato dextrose agar (PDA) media containing 30, 60 and 90 (mg/kg) BaP for adaptation of the fungal strains. Removing of BaP was measured after 45 days growth of the fungal colonies under different concentrations of BaP pollution in PDA media and soil. Peroxidase and catalase activity, and protein content were compared in the fungi growing in BaP-polluted media and control ones. The results showed that all the isolated fungi were able to growth in the BaP containing media and could remove BaP from the media. The highest removal efficiency was determined for Fusarium acuminatum (93%). Similar data obtained when the fungus used for bioremediation of BaP-polluted soil (91%). Total protein content and enzymatic activity (Peroxidase and Catalase) were increased with increasing of BaP pollution. The highest catalase activity was measured in F. acuminatum growing in the media containing 90 mg/kg BaP (2.2×10-2 unit/mg protein) and the highest (5× 10-3 unit/mg protein) peroxidase activity for Alternaria alternata. Protein content in the fungi was increased with increasing of BaP pollution. In F. acuminatum, the lowest amount of total protein was observed in the control sample (1×10-3 mg/g FW) and the highest amount was belonging to the group treated by 90 mg/kg BaP (7.5×10-3 mg/g FW). It concluded that F. acuminatum showed the highest catalase activity, highest total protein content and also the highest BaP removal efficiency from both BaP-polluted media and soils.

Optimizing of protease purification from bacillus cereus td5b by ammonium sulfate precipitation

The aim of this research was to optimize the protease purification from Bacillus cereus TD5B by ammonium sulfate precipitation. The material used was protease enzyme from Bacillus cereus TD5B. It was produced and purified in a liquid medium using ammonium sulfate precipitation. The optimization was performed using various levels (50, 60, 70 and 80 %) of ammonium sulfate as precipitation agent of crude microbial enzymes. Separation process of precipitate and supernatant was conducted by 3,500 rpm centrifugation for 30 min at 4°C. The specific activity of protease enzyme, and protein content was observed on precipitate and supernatant of each levels treatment of the ammonium sulfate. Based on specific enzyme activity and protein content, the measurement showed that 70 % ammonium sulfate level treatment has the highest specific enzyme activity (78.296 U/mg) and highest protein content (2.689 mg/mL). This specific enzyme activity was three times higher compared to specific enzyme activity before purification treatment (24.12 U/mg). In conclusion, the purification of crude Bacillus cereus TD5B enzyme by ammonium sulfate purification can increase the specific enzyme activity.

Evidence of Increased Prefrontal Cortex Inflammation and Amyloid Precursor Protein Processing in a Translational Swine Model of Heart Failure with Preserved Ejection Fraction

The oligomerization of amyloid beta peptides (Aβ), a histopathological hallmark of diseases such as Alzheimer's, may be elevated in cardiogenic dementias coincident with heart failure (HF). BACE1, the rate‐limiting protease enzyme in the amyloidogenic process, initiates the processing of amyloid precursor protein (APP) leading to the production of Aβ. Understanding the mechanistic regulation of BACE1 may be critical for improving treatment efficacy in heart failure patients exhibiting cardiogenic dementia. The purpose of this study was to examine the molecular mechanisms of cardiogenic dementia in a novel swine model of HF with preserved ejection fraction (HFpEF) exhibiting comorbidities including obesity and metabolic syndrome. Female Ossabaw pigs were randomly divided into two groups; control (CON, n=4) and Western diet‐induced obesity/metabolic syndrome with pressure overload‐induced HFpEF (HF, n=4). The HF group was placed on a Western diet (starting at 3 mo. of age) and aortic‐banded (at 6 mo. of age) prior to euthanasia (at 12 mo. old). At euthanasia, prefrontal cortex and hippocampus regions were collected for Western blot analysis. Total body mass was increased (CON=42±2 vs. HF=72±4 Kg; p&lt;0.01), but brain mass was decreased (CON=110±3 vs. HF=92±2 g; p&lt;0.01) in the HF group. The HF group exhibited greater prefrontal cortex total APP (p&lt;0.01), BACE1 (p&lt;0.01) and CTF protein levels (p=0.01), indicative of increased APP processing. This was accompanied by increased markers of chronic stress and neuroinflammation, specifically phosphorylation of JNK (p&lt;0.01) and phosphorylation of ERK (p&lt;0.01). Aberrant control of the insulin‐signaling pathway was also observed in the prefrontal cortex via increases in the phosphorylation of Akt serine 473 (p&lt;0.01) and insulin‐degrading enzyme protein content (p=0.03) in the HF group. Similar results were observed in the hippocampus for increases in APP processing (increased APP and CTF content), neuroinflammation (phosphorylation of JNK and ERK). However, there were no changes in BACE1 content or the insulin‐signaling pathway, highlighting the importance of examining regional differences in the brain. These data reveal that inflammatory stress and aberrant control of insulin signaling may be implicated in the pathogenic regulation of amyloid beta peptide metabolism in an experimental, large animal setting of HFpEF, and potentially contribute to the cardiogenic dementia often observed in HF patients.Support or Funding InformationNIH R01 HL112998, PI: CAENIH K01 HL125503 – PI, PadillaAmerican Heart Association #16POST2776005, PI: TDOUniversity of Missouri Research Board – PI's, Emter and RectorAlzheimer Society of Brant, Haldimand, Norfolk, Hamilton, and Halton, PI: REKMThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Modulation of arsenic-induced oxidative stress and protein metabolism by diphenyleneiodonium, 24-epibrassinolide and proline in Glycine max L.

AbstractArsenic (As)-toxicity is a major constraint for crop production. The present study was intended to examine the comparative ameliorative effects of diphenyleneiodonium (DPI), 24-epibrassinolide (EBL) and proline (Pro) on As-stress inGlycine maxL. Seeds ofGlycine maxL. were subjected to As (100 µM) singly, and together with DPI (10 µM), EBL (0.5 µM) or Pro (10 mM), for five days, and were then analyzed. Experimental results showed that As treatment caused a substantial fall in growth traits like germination percentage, radicle length and dry mass, which was accompanied by As accumulation. Additionally, As application also revealed reduced viability, total protein content and activities of antioxidative enzymes (superoxide dismutase, catalase and ascorbate peroxidase), while it increased the levels of total sugar, proline and oxidative stress markers such as electrolyte leakage, reactive oxygen species, lipid oxidized products, protein carbonyls and hydroperoxides, Amadori and Maillard reaction products, malondialdehyde-/4-hydroxy-2-nonenal-protein adducts, protease and proteasome. Isozymes of antioxidative enzymes were also observed to be altered considerably under As-stress. Impressively, DPI, EBL and Pro played their role as protective agents, hence caused enhanced growth and reduced As accumulation. These protective chemicals also improved the viability, accruals of total protein, total sugar and endogenous proline, and activities of antioxidants, while they reduced the levels of oxidative stress markers. Our findings demonstrated the involvement of DPI, EBL and Pro in As-stress tolerance inGlycine maxL. Further, Pro appears to be superior to DPI and EBL, in alleviating As-induced responses inGlycine maxL.

Retraction of articles by Dr M. Aramli.

The below articles published online on Wiley Online Library (wileyonlinelibrary.com) have been retracted by agreement between the submitting author, Mohammad Sadegh Aramli (2015, 2015a, 2015b), the Editor-in-Chief, Michel Goldberg, and Blackwell Verlag GmbH. After a thorough investigation, there is strong evidence to indicate that the peer review of these papers was compromised. The identities of the reviewers were unable to be verified, and it is believed that these papers were accepted based on recommendations from reviewers not suitably qualified. REFERENCE Aramli, M. S. (2015). Study of enzyme activities and protein content of beluga (Huso huso) semen before and after cryopreservation. Journal of Animal Physiology and Animal Nutrition, 99, 13-16. https://doi.org/10.1111/jpn.12199 Aramli, M. S., Habibi, E., Aramli, S., & Nouri, H. A. (2015b). Determination of some blood and seminal plasma ions in the beluga, Huso huso (Linnaeus, 1758). Journal of Animal Physiology and Animal Nutrition, 99, 17-20. https://doi.org/10.1111/jpn.12225 Aramli, M. S., Kalbassi, M. R., & Gharibi, M. R. (2015a). Effects of multiple collections on spermatozoa quality of Persian sturgeon, Acipenser persicus: Motility, density and seminal plasma composition. Journal of Animal Physiology and Animal Nutrition, 99, 66-72. https://doi.org/10.1111/jpn.12212.

Vitamin C attenuates biochemical and genotoxic damage in common carp (Cyprinus carpio) upon joint exposure to combined toxic doses of fipronil and buprofezin insecticides

In the present study, potential protective role of Vitamin C (l-ascorbic acid) was investigated in aquaria acclimated common carp (Cyprinus carpio) following exposure for 96 h to combined toxic doses of fipronil (FP) and buprofezin (BPFN) insecticides in combination (FP: 200 μg/L; 4.57 × 10−7 mol/L and BPFN: 50 mg/L; 1.64 × 10−4 mol/L). At end of 96 h exposure, fish were supplemented with low (25 mg/L) and high (50 mg/L) doses of Vitamin C, added once daily to aquaria water for continuous three weeks. Appropriate control groups were run in parallel. Fish behavior was monitored throughout for signs of toxicity. At completion of experiments, liver, kidney, brain and gills were excised for toxicity assessment and possible remediation by the Vitamin C through biochemical determination of reactive oxygen species (ROS), thiobarbituric acid reactive substances or TBARS, reduced glutathione (GSH) and total protein content, levels of catalase (CAT), superoxide dismutase (SOD) and peroxidase (POD), and the Comet assay. Hepatosomatic index (HSI), condition factor (CF), survival rate (SR), and combination index (CI) were also determined. Data were compared statistically at p < 0.05. Results showed significant behavioral and biochemical alterations, and DNA damage in the fish group exposed to FP and BPFN in combination. In fish groups supplemented with Vitamin C following FP and BPFN treatment, significant alleviation in tissue damage and toxic effects was represented by substantial decreases in ROS and TBARS production (p < 0.001), along with a concomitant significant increase in the survival rate, GSH and total protein content, HSI, CF, and activities of SOD, CAT and POD enzymes (p < 0.001). Mean tail length of comet and percent tail DNA decreased significantly (p < 0.001), which indicated amelioration of DNA damage. The study concludes that Vitamin C is an effective remedial treatment against FP and BPFN-induced damage in exposed fish.

The Effects of Ibogaine on Uterine Smooth Muscle Contractions: Relation to the Activity of Antioxidant Enzymes

Ibogaine is an indole alkaloid originally extracted from the root bark of the African rainforest shrub Tabernanthe iboga. It has been explored as a treatment for substance abuse because it interrupts drug addiction and relieves withdrawal symptoms. However, it has been shown that ibogaine treatment leads to a sharp and transient fall in cellular ATP level followed by an increase of cellular respiration and ROS production. Since contractile tissues are sensitive to changes in the levels of ATP and ROS, here we investigated an ibogaine-mediated link between altered redox homeostasis and uterine contractile activity. We found that low concentrations of ibogaine stimulated contractile activity in spontaneously active uteri, but incremental increase of doses inhibited it. Inhibitory concentrations of ibogaine led to decreased SOD1 and elevated GSH-Px activity, but doses that completely inhibited contractions increased CAT activity. Western blot analyses showed that changes in enzyme activities were not due to elevated enzyme protein concentrations but posttranslational modifications. Changes in antioxidant enzyme activities point to a vast concentration-dependent increase in H2O2 level. Knowing that extracellular ATP stimulates isolated uterus contractility, while H2O2 has an inhibitory effect, this concentration-dependent stimulation/inhibition could be linked to ibogaine-related alterations in ATP level and redox homeostasis.

Impact of training state on fasting-induced regulation of adipose tissue metabolism in humans.

Recruitment of fatty acids from adipose tissue is increased during fasting. However, the molecular mechanisms behind fasting-induced metabolic regulation in human adipose tissue and the potential impact of training state in this are unknown. Therefore the aim of the present study was to investigate 1) fasting-induced regulation of lipolysis and glyceroneogenesis in human adipose tissue as well as 2) the impact of training state on basal oxidative capacity and fasting-induced metabolic regulation in human adipose tissue. Untrained [maximal oxygen uptake (V̇o2max) < 45 ml·min-1·kg-1] and trained subjects (V̇o2max > 55 ml·min-1·kg-1) fasted for 36 h, and abdominal subcutaneous adipose tissue biopsies were obtained 2, 12, 24, and 36 h after a standardized meal. Adipose tissue oxidative phosphorylation complexes, phosphoenolpyruvate carboxykinase, and pyruvate dehydrogenase (PDH)-E1α protein as well as PDH kinase (PDK) 2, PDK4, and PDH phosphatase 2 mRNA content were higher in trained subjects than in untrained subjects. In addition, trained subjects had higher adipose tissue hormone-sensitive lipase Ser660 phosphorylation and adipose triglyceride lipase protein content as well as higher plasma free fatty acid concentration than untrained subjects during fasting. Moreover, adipose tissue PDH phosphorylation increased with fasting only in trained subjects. Taken together, trained subjects seem to possess higher basal adipose tissue oxidative capacity as well as higher capacity for regulation of lipolysis and for providing substrate for glyceroneogenesis in adipose tissue during fasting than untrained subjects. NEW & NOTEWORTHY This study shows for the first time higher protein content of lipolytic enzymes and higher oxidative phosphorylation protein in adipose tissue from trained subjects than from untrained subjects during fasting. Furthermore, trained subjects had higher capacity for adipose tissue glyceroneogenesis than untrained subjects.

Diclofenac exposure alter the expression of PXR and its downstream target genes in mosquito fish (Gambusia affinis)

As one of widely used drugs, Diclofenac (DCF) recently has been universally detected in aquatic environment and some negative effects derived from DCF exposure to mammals have been also reported. However, studies about its potential deleterious effects on non-target organisms like fish still require more investigation. In this study an ubiquitous small freshwater invader species in Southern of China, mosquito fish (Gambusia affinis), was employed as test organism. We firstly cloned the crucial partial sequences of nucleus transcriptional factor related genes pregnane X receptor (PXR) and its downstream genes, including P-glycoprotein (P-gp), cytochrome 3A (CYP3A), multidrug resistance protein 2 (MRP2), glutathione peroxidase (GPx) and thioredoxin reductase (TXR) in mosquito fish. The phylogenetic trees of PXR, CYP3A and MRP2 were constructed based on their deduced amino acids sequences, respectively. Phylogenetic trees and blast results showed a high similarity between G. affinis and other killifish species, such as Xiphophorus maculatus. The transcriptional expression of these genes mentioned above and partly related enzymes/proteins activities were then measured under the exposure of environmentally relevant concentrations of DCF (from 0.5μgL−1 to 500μgL−1) for 24h and 168h. Results showed that the mRNA expression of PXR, CYP3A, P-gp and TXR showed dramatic induction under DCF exposure, exhibiting an obvious time-effect relationship with the extend of exposure time. In terms of enzyme activity and protein content, no dramatic changes as in transcription were observed. Western blotting showed PXR protein increased at 24h but decreased at 168h with the increasing of DCF concentration, displaying a dose-effect relationship to some extent. GPX activity was continuously induced both at 24h and 168h, exhibiting a good consistency with the performance of GPX gene. GSSH/T-GSH increased in all treatments. Overall, DCF had traceable effects on the expression of PXR and its downstream target genes in mosquito fish.

Changes in the secretory activity of organs producing noradrenaline upon inhibition of its synthesis in neonatal rat brain

The study is devoted to investigating the humoral interaction of the organs that are major sources of the morphogenetic factor noradrenaline in the general circulation system of rats in the neonatal period of development. The change in the synthesis of noradrenaline in the central and peripheral organs when its synthesis in the brains of neonatal rats is switched off was considered as one of the mechanisms of such interaction. For this purpose, the level of gene expression and the content of enzymes involved in the synthesis of noradrenaline—tyrosine hydroxylase (TH) and dopamine-β-hydroxylase (DBH)—was determined in the brain, adrenal glands, and the organ of Zuckerkandl. The synthesis of noradrenaline after destruction of noradrenergic neurons was switched off by injecting anti-DBH-saporin-antibodies against dopamine-β-hydroxylase conjugated to the cytotoxin saporin into the cerebral ventricles of neonatal rats. The content of TH and DBH mRNA in organs was determined by realtime PCR, and the protein content of enzymes was determined by Western blotting. It is shown that, 48 h after switching off the synthesis of noradrenaline in the brain of 2-day-old animals, a sharp increase in TH and DBH mRNA content in adrenal glands and the organ of Zuckerkandl was observed, although the content of both enzymes was retained at the control level. These data indicate a compensatory enhancement of expression of the genes encoding noradrenaline synthesis enzymes at the transcriptional level as a result of humoral interaction of central and peripheral sources of noradrenaline in the neonatal period of ontogeny.

Activity of catalase enzyme in Paulownia tomentosa seeds during the process of germination after treatments with low pressure plasma and plasma activated water

In this work we present results of two significantly different types of plasma treatment on Paulownia tomentosa Steud. seeds. In the first type, seeds were directly treated in low‐pressure plasma and then imbibed with distilled water. In the second type, an atmospheric pressure plasma was used for obtaining plasma activated water (PAW) which is then used for imbibition of seeds. The CAT activity and protein content is evaluated during 4 d following the imbibition process, i.e., immediately after the phytochrome activation and in the 3 subsequent days. Comparison of results of treated seeds to the control group allows to correlate the enzyme activity and protein content during the initial stages of germination with plasma treatment types and treatment conditions.