Significance of the Wnt canonical pathway in radiotoxicity via oxidative stress of electron beam radiation and its molecular control in mice.

PURPOSE: Radiation triggers cell death events through signaling proteins, but the combined mechanism of these events is unexplored The Wnt canonical pathway, on the other hand, is essential for cell regeneration and cell fate determination. AIM: The relationship between the Wnt pathway's response to radiation and its role in radiotoxicity is overlooked, even though it is a critical molecular control of the cell. The Wnt pathway has been predicted to have radioprotective properties in some reports, but the overall mechanism is unknown. We intend to investigate how this combined cascade works throughout the radiation process and its significance over radiotoxicity. MATERIALS AND METHODS: Thirty adult mice were irradiated with electron beam radiation, and 5 served as controls. Mice were sacrificed after 24 h and 30 days of irradiation. We assessed DNA damage studies, oxidative stress parameters, mRNA profiles, protein level (liver, kidney, spleen, and germ cells), sperm viability, and motility. OBSERVATION: The mRNA profile helps to understand how the combined cascade of the Wnt pathway and NHEJ work together during radiation to combat oxidative response and cell survival. The quantitative examination of mRNA uncovers unique critical changes in all mRNA levels in all cases, particularly in germ cells. Recuperation was likewise seen in post-30 day's radiation in the liver, spleen, and kidney followed by oxidative stress parameters, however not in germ cells. It proposes that reproductive physiology is exceptionally sensitive to radiation, even at the molecular level. It also suggests the suppression of Lef1/Axin2 could be the main reason for the permanent failure of the sperm function process. Post-irradiation likewise influences the morphology of sperm. The decrease in mRNA levels of Lef1, Axin2, Survivin, Ku70, and XRCC6 levels suggests radiation inhibits the Wnt canonical pathway and failure in DNA repair mechanisms in a coupled manner. An increase in Bax, Bcl2, and caspase3 suggests apoptosis activation followed by the decreased expression of enzymatic antioxidants. CONCLUSION: Controlled several interlinked such as the Wnt canonical pathway, NHEJ pathway, and intrinsic apoptotic pathway execute when the whole body is exposed to radiation. These pathways decide the cell fate whether it will survive or will go to apoptosis which may further be used in a study to counterpart and better comprehend medication focus on radiation treatment.

Enhanced cellular uptake, transport and oral bioavailability of optimized folic acid-loaded chitosan nanoparticles.

Folic acid is a synthetic form of folate widely used for food fortification. However, its bioavailability is limited due to its inherent instability at several conditions. Therefore, a suitable encapsulation system is highly required. In the present study, the fabrication condition for folic acid-loaded chitosan nanoparticle (FA-Chi-NP) was optimized and then subjected to characterization. The optimized formulation had the particle size, zeta potential, and encapsulation efficiency of 180 nm, +52 mV, and 90%, respectively. In vitro release profile showed a controlled release of folic acid from the nanoparticles. Treatment of Caco-2 cells with the formulation showed no adverse effects based on MTT and LDH assays, and also, the cellular uptake was significantly higher after 2 h compared to free folic acid. Further, the oral administration of rats with FA-Chi-NPs (1 mg/kg BW) increased the plasma level of both folic acid (3.2-fold) and its metabolites such as tetrahydrofolate (2.3-fold) and 5-methyltetrahydrofolate (1.6-fold) significantly compared to free folic acid. In a bio-distribution study, duodenum and jejunum were found to be the primary sites for absorption. These findings suggest that chitosan may be a promising carrier for the delivery of folic acid and, therefore, could be exploited for various food applications.

Bacopa monniera extract mitigates isoproterenol-induced cardiac stress via Nrf2/Keap1/NQO1 mediated pathway.

The present study was aimed to investigate the effect of standardised hydroalcoholic extract of Bacopa monniera (BME) against isoproterenol (ISO) induced cardiac stress. Isoproterenol (85 mg/kg body weight) was administered intraperitoneally to induce cardiac stress in rats. Bacopa monniera extract (BME75 and 150 mg/kg) was orally administered for 21 days followed by ISO on 22nd and 23rd experimental days. ISO caused significant cardiac damage, which was concomitant with increased apoptosis and attenuated expressions of Nrf2, HO-1, and regulating apoptotic protein expressions of Bax, Bcl2 and NOS2. Treatment with BME in rats significantly improved cardiac dysfunction by maintaining cardiac rhythm, myocardial integrity. Decreased oxidative stress by restored expressions of Nrf2, NQO1 and HO-1 followed by elevating antioxidant enzymes and total glutathione levels. Our present results suggest that the BME treatment strengthening the endogenous defence system through Nrf2 modulation and played a key role against cardiac oxidative stress induced by ISO in rats.

Ameliorative Effect of Hesperidin Against Motion Sickness by Modulating Histamine and Histamine H1 Receptor Expression.

Motion sickness (MS) is the visceral discomfort caused due to contradicting visual and vestibular inputs to the brain leading to nausea and vomiting. Sensory conflict theory which proves histamine elevations as the primary reason for MS provides a path for an effective pharmaco-therapy. We aimed to evaluate the anti-MS effect of hesperidin (HSP) by modulating histamine and histamine receptor H1 (HRH1) expression. The inhibitory effect of HSP on histamine release was studied in KU812 cells treated with 10 µM calcium ionophore. The in vivo anti-MS effect of HSP was evaluated in Balb/c mice. Thirty six mice were divided into six groups namely, normal control (NC, no rotation), hesperidin at 80 mg/kg body weight control (HSP80, no rotation), motion sickness (MS, rotation induced), dimenhydrinate (Standard drug) at 20 mg/kg body weight + rotation (STD + MS), hesperidin at 40 mg/kg body weight + rotation (HSP40 + MS) and hesperidin at 80 mg/kg body weight + rotation (HSP80 + MS). Hypothalamus and brainstem samples were analysed for histamine levels and HRH1 expression by RT-PCR, Western blot and immunohistochemistry analysis. Calcium ionophore treated KU812 cells significantly increased histamine release when compared to control cells. Pre-treatment with HSP inhibited histamine, HRH1 mRNA and protein expression. Histamine, HRH1 mRNA and protein expression in hypothalamus and brainstem samples of MS group increased significantly when compared to the NC group. Pre-treatment with HSP significantly reduced histamine, HRH1 mRNA and protein expression. Thus, indicating that HSP has a potent anti- MS effect by decreasing the elevated levels of histamine, HRH1 mRNA and protein expression in hypothalamus and brainstem regions.

In-silico therapeutic investigations of arjunic acid and arjungenin as an FXR agonist and validation in 3T3-L1 adipocytes.

The present study was to illustrate the agonistic property of arjungenin and arjunic acid towards farnesoid X receptor protein (FXR).The pharmacokinetic properties like molecular interactions, absorption, distribution, metabolism, elimination and toxicity (ADMET) of the ligands were checked through in-silico studies. Protein-ligand docking was carried out using autodock software. Molecular docking analysis confirmed strong binding energy and interaction of arjungenin and arjunic acid with the target protein and the ADMET profiles identified for both compounds were promising.Further in vitro studies were performed in 3T3-L1 adipocyte to verify the agonistic property of arjungenin and arjunic acid. Oil red O staining was done to check differentiation induction. Adiponectin, leptin, triglycerides and total cholesterol levels were quantified. The mRNA expression of FXR, Cyp7a1, PPAR-γ and SREBP-1c were quantified using fluorescent real-time PCR. Cytotoxicity assay was confirmed that up to 150 µM concentration there is no significant cell death on treatment with arjunic acid and arjungenin. Treatment with arjungenin and arjunic acid confirms increased differentiation of the cells with significant (P < 0.05) increase in adiponectin (118.07% and 132.92%) and leptin (133.52% and 149.74%) protein levels compared to the negative control group. After treatment with arjungenin and arjunic acid in 3T3-L1 preadipocytes the mRNA expression of FXR, PPAR-γ and SREBP-1c were significantly (P < 0.01) increased and cyp7a1 was significantly (P < 0.01) decreased when compared with the negative control group. Overall, our results suggest that arjungenin and arjunic acid acts as an FXR agonist and may be useful for rational therapeutic strategies as a novel drug to treat cholesterol mediated metabolic syndrome and insulin resistance.

Influence of parboiling of red paddy varieties by simple hot soaking on physical, nutrient, phytochemical, antioxidant properties of their dehusked rice and their mineral, starch, and antioxidant's bioaccessibility studies.

The objective of this study was to know about the effect of simple parboiling on physical properties, proximate composition, phenolic compounds, and antioxidant activity.  These were  studied in raw and parboiled paddy varieties as well as bioaccessibility of specific nutrients (minerals, starch, and antioxidants). The pigmented rice paddy varieties such as Jyothi, Meter & Athikaraya were parboiled by hot soaking treatment after soaking for 2, 2½, and 3 hr. Athikaraya rice showed high protein, low ash in raw and parboiled than other varieties. Amylose equivalents of all three varieties exhibited in the range of 24%-27% (d.b). Whereas, the soluble amylose content showed 12.7, 8.7, and 7.7% (d.b) in Athikaraya, Meter and Jyothi rice varieties, respectively. Jyothi rice showed less cooking time and more cooking volume. Athikaraya showed high phenolic content and antioxidant properties compared to other two varieties. The dialysability of minerals, starch, and antioxidants were increased due to simple hot soaking parboiling. PRACTICAL APPLICATIONS: The present study has the significance in assessing the variation among different pigmented rice varieties after parboiling by simple hot soaking with various soaking periods. The industrial advantage of this method is, there is no usage of boiler for cooking the paddy by steam, which was economically better. The information gained by this study might be beneficial for consumers and suppliers regarding the quality of the selected pigmented rice varieties with respect to nutrient composition, antioxidant activities, and bioaccessibility of minerals, starch, and antioxidants. Ultimately, the present study can lead to better appreciation of pigmented rice and assist food processors in selecting rice variety with unique characteristics for specialty food preparations.

Terminalia arjuna extract and arjunic acid mitigate cobalt chloride-induced hypoxia stress-mediated apoptosis in H9c2 cells.

Arjunic acid (AA) is one of the major active component of Terminalia arjuna known for its health benefits. In the present study, we evaluated cardioprotective potential of Terminalia arjuna extract (TAE) and AA against cobalt chloride (CoCl2)-induced hypoxia damage and apoptosis in rat cardiomyocytes. TAE (50 µg/ml) and AA (8 µg/ml) significantly (p < 0.001) protected H9c2 cells as evidenced by cell viability assays against CoCl2 (1.2 mM)-induced cytotoxicity. TAE and AA pretreatments protected the cells from oxidative damage by decreasing the generation of free radicals (ROS, hydroperoxide, and nitrite levels). TAE and AA pretreatments retained mitochondrial membrane potential by alleviating the rate of lipid peroxidation induced by CoCl2 treatment. TAE and AA pretreatments elevated antioxidant status including phase II antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase) and total glutathione levels against CoCl2-induced oxidative stress. Further immunoblotting studies confirmed anti-apoptotic effects of TAE and AA by alleviating the phosphorylation of JNK and c-jun and also by regulating protein expression levels of Bcl2, Bax, caspase 3, heat shock protein-70, and inducible nitric oxide synthase. Overall, our results suggest that both the extract and the active component exhibit antioxidant and anti-apoptotic defense against CoCl2-induced hypoxic injury.

Chemical composition, antioxidant potential, macromolecule damage and neuroprotective activity of Convolvulus pluricaulis.

Herbal medicines are known to mitigate radical induced cell damage. Hence identification and scientific validation of herbal medicines contribute to better use in Ayurvedic/Unani research. In the present study, we investigated antioxidant and anti-apoptotic properties of Convolvulus pluricaulis (C. pluricaulis). C. pluricaulis exhibited antioxidant potential evident by free radical scavenging activities. C. pluricaulis pretreatment inhibited H2O2 induced macromolecule damage such as plasmid DNA damage and AAPH induced oxidation of bovine serum albumin and lipid peroxidation of rat hepatic tissues. Further to identify the neuroprotective properties of C. pluricaulis, SHSY5Y cells were treated with H2O2 with or without pretreatment of C. pluricaulis. The C. pluricaulis pretreatment at 50 µg/ml dose exhibited 50% cell survival against 100 µM H2O2 challenge for 24 h and it also decreased the lactate dehydrogenase leakage. Further C. pluricaulis pretreatment restored and regulated the antioxidant and apoptosis markers such as SOD, CAT, p53, and caspase-3 and inhibited, reactive oxygen species generation and depolarization of the mitochondrial membrane. C. pluricaulis possess a high content of flavonoids and polyphenols and GC-MS and FTIR analysis showed a wide variety of compounds which may contribute to the observed effects.

Anxiolytic actions of Nardostachys jatamansi via GABA benzodiazepine channel complex mechanism and its biodistribution studies.

Nardostachys jatamansi has profound applications against pharmacological interventions and is categorized as a hypno-sedative drug according to Ayurveda. In the present study probable mechanism of anxiolytic action of Nardostachys jatamansi extract (NJE) was studied using behavioral anxiolytic tests (Elevated plus maze, Open field test, Light dark box test, and Vogel's conflict test) in mice. Mice were treated orally with NJE (250 mg/kg) for 3, 7 and 14 days or diazepam (1 mg/kg) followed by behavioral assessment and estimation of monoamine neurotransmitters, GABA, and antioxidant enzymes. Treatment of mice for 7 days caused an increase in time spent in open arms in elevated plus maze, number of line crossings in open field test, increased time spent in lit compartment of light-dark box test, an increase in number of licks made and shocks accepted in Vogel's conflict test, with results comparable to diazepam and this treatment also caused a significant increase in monoamine neurotransmitters and GABA in brain and tissue antioxidant parameters. Co-treatment of NJE with flumazenil (GABA-benzodiazepine antagonist; 0.5 mg/kg i.p) or picrotoxin (GABAA gated chloride channel blocker; 1 mg/kg i.p) caused a blockage/antagonised anxiolytic actions of NJE by causing a significant reduction in time spent in open arms of elevated plus maze, an decrease in number of line crossing in open field test and also number of shocks and licks accepted in Vogel's conflict test. Further, NJE was radiolabelled with technetium99m at their hydroxyl groups following which purity as well as in vivo and in vitro stability of radiolabelled formulations was evaluated. The blood kinetics and in vivo bio-distribution studies were carried out in rabbits and mice respectively. Labeled formulation was found to be stable in vitro (96 to 93% stability) and in vivo (96 to 92% stability). The labeled compound was cleared rapidly from blood (within 24 h) and accumulated majorly in kidneys (11.65 ± 1.33), liver (6.07 ± 0.94), and blood (4.03 ± 0.63) after 1 h. However, a small amount was observed in brain (0.1 ± 0.02) probably because of its inability to cross blood-brain barrier. These results highlight biodistribution pattern of NJE, and also indicated that a 7-day treatment with NJE produced significant anxiolytic effects in mice and also a significant increase in brain monoamine and GABA neurotransmitter levels and suggests that anxiolytic effects of NJE are primarily and plausibly mediated by activating GABAergic receptor complex.

Restorative effect of l-Dopa treatment against Ochratoxin A induced neurotoxicity.

The toxic effects of Ochratoxin A (OTA), a fungal secondary metabolite of the genera Aspergillus and Penicillium with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) a Parkinson inducing drug were investigated to evaluate the neurotoxic effects exerted by OTA. OTA is known to contaminate food and feedstuff leading to a wide range of toxicity like nephrotoxicity, hepatotoxicity, and immunotoxicity. However, due to the dearth of available information on the possible mechanisms of OTA neurotoxicity and neurodegeneration the current study was undertaken. Hence, in this study, we examined the neurotoxic effects and the possible mechanism of action of neurodegeneration by OTA toxicity on mice brain by conducting a battery of behavioural studies and reviewing neurotransmitter levels and neuronal apoptotic pathways. Further, they were treated with l-Dopa, a precursor of dopamine (DA) to explore its ameliorative effects against OTA. The results of behavioural studies like gait analysis, spontaneous activity, cylinder test and pole test showed that OTA exhibits Parkinsonian physiognomies which were stabilized with l-Dopa treatment. Also, OTA toxicity showed insults on neurotransmitter levels and general brain function parameters that were normalized with l-Dopa treatment. The results of the present study suggest that OTA promotes neurodegeneration by targeting neuronal pathway leading to the development of Parkinson's diseases.

Attenuation of cytotoxicity induced by tBHP in H9C2 cells by Bacopa monniera and Bacoside A.

Cardiovascular diseases are one of the major global health issues leading to morbidity and mortality across the world. In the present study Bacopa monniera and its major bioactive component, Bacoside A (Bac-A) was used to evaluate its cytoprotective property in H9C2 cardiomyocytes against tBHP (150 µM) induced ROS-mediated oxidative stress and apoptosis. Our results implicate that pre-treatment with hydroalcoholic extract of Bacopa monniera (BME) and Bac-A (125 µg/ml and 6 µg/ml respectively) significantly restored oxidative stress by scavenging the free radicals and also elevated phase II antioxidant defensive enzymes such as (SOD, CAT, GR, GPx and GSH). Membrane integrity was estimated by MMP and LDH assays and found 89 and 72% of the protective effect. Further immunoblotting studies confirmed anti-apoptotic effects by regulating protein expression like Bcl2 was up-regulated to 99 and 85% and Bax was down-regulated to 122 and 181%, iNOS by 154.38 and 183.45% compared to tBHP (277.48%) by BME and Bac-A. BME and Bac-A exerts cytoprotective efficacy by attenuation of ROS generated through oxidative stress by an increase in the concentration of antioxidant enzymes and sustain membrane integrity which leads to restoring the damage caused by tBHP.

Pelargonidin Modulates Keap1/Nrf2 Pathway Gene Expression and Ameliorates Citrinin-Induced Oxidative Stress in HepG2 Cells.

Pelargonidin chloride (PC) is one of the major anthocyanin found in berries, radish and other natural foods. Many natural chemopreventive compounds have been shown to be potent inducers of phase II detoxification genes and its up-regulation is important for oxidative stress related disorders. In the present study, we investigated the effect of PC in ameliorating citrinin (CTN) induced cytotoxicity and oxidative stress. The cytotoxicity of CTN was evaluated by treating HepG2 (Human hepatocellular carcinoma) cells with CTN (0-150 µM) in a dose dependent manner for 24 h, and the IC50 was determined to be 96.16 µM. CTN increased lactate dehydrogenase leakage (59%), elevated reactive oxygen species (2.5-fold), depolarized mitochondrial membrane potential as confirmed by JC-1 monomers and arrested cell cycle at G2/M phase. Further, apoptotic and necrotic analysis revealed significant changes followed by DNA damage. To overcome these toxicological effects, PC was pretreated for 2 h followed by CTN exposure for 24 h. Pretreatment with PC resulted in significant increase in cell viability (84.5%), restored membrane integrity, reactive oxygen species level were maintained and cell cycle phases were normal. PC significantly up-regulated the activity of detoxification enzymes: heme oxygenase 1 (HO-1), glutathione transferase, glutathione peroxidase, superoxide dismutase and quinone reductase. Nrf2 translocation into the nucleus was also observed by immunocytochemistry analysis. These data demonstrate the protective effect of PC against CTN-induced oxidative stress in HepG2 cells and up-regulated the activity of detoxification enzyme levels through Keap1/Nrf2 signaling pathway.

Patulin Induced Oxidative Stress Mediated Apoptotic Damage in Mice, and its Modulation by Green Tea Leaves.

BACKGROUND: The present study demonstrates the antioxidant and hepatic protective effects of Green tea leaves (GTL). METHODS: The serum level of aspartate aminotransferase and alanine aminotransferase was analyzed. The liver antioxidant enzymes such as SOD, CAT, GPx, GR, GSH, lipid peroxidation and protein carbonyls, ROS content were estimated. The histology of liver tissue was observed and the protein expression of SOD, CAT, Caspase-3 and p53 was investigated by Western blotting. RESULTS: Effectiveness of GTL extract in preventing patulin induced liver damage showed significant reduction in serum ALT and AST to 19% and 85% respectively, the increase in antioxidant levels and lipid peroxidation products with patulin treatment were also reduced with GTL supplementation. The patulin induced increase in hepatic protein carbonyls was significantly reduced by 141-111% with 100 and 200 mg/kg b.wt GTL and in ROS was significantly reduced by 171-140% with 100 and 200 mg/kg b.wt GTL administration respectively. Also showed protection against hepatic tissue damage and protein expression in mice. CONCLUSION: This study showed remarkable antioxidant and hepatic protective effects of GTL.

Cytoprotective propensity of green tea polyphenols against citrinin-induced skeletal-myotube damage in C2C12 cells.

The mycotoxin citrinin, is produced by several species of Penicillium, Aspergillus and Monascus, and is capable of inducing cytotoxicity, oxidative stress and apoptosis. The aim of the present study was to investigate the effect of citrinin in mouse skeletal muscle cells (C2C12) and to overcome the cellular adverse effects by supplementing green tea extract (GTE) rich in polyphenols. C2C12 myoblasts were differentiated to myotubes and were exposed to citrinin in a dose dependent manner (0-100 µM) for 24 h and IC50 value was found to be 100 µM that resulted in decreased cell viability, increased LDH leakage and compromised membrane integrity. Mitochondrial membrane potential loss, increased accumulation of intracellular ROS and sub G1 phase of cell cycle was observed. To ameliorate the cytotoxic effects of CTN, C2C12 cells were pretreated with GTE (20, 40, 80 µg/ml) for 2 h followed by citrinin (100 µM) treatment for 24 h. GTE pretreatment combated citrinin-induced cytotoxicity and oxidative stress. GTE at 40 and 80 µg/ml significantly promoted cell survival and upregulated antioxidant enzyme activities (CAT, SOD, GPx) and endogenous antioxidant GSH, while the gene and protein expression levels were significantly restored through its effective antioxidant mechanism. Present study results suggested the antioxidant properties of GTE as a herbal source in ameliorating the citrinin-induced oxidative stress.

Anti-inflammatory and anti-oxidant effects of Cardamom (Elettaria repens (Sonn.) Baill) and its phytochemical analysis by 4D GCXGC TOF-MS.

Elleteria repens is a large cardamom used in the culinary preparations. In the present study, we have evaluated in vitro antioxidant and free radical scavenging activities E. repens hexane extract (ERH) exhibited DPPH and metal chelating activity with IC50 values of 464±28.3µg/ml, 199±7.2µg/ml whereas the reducing power and antioxidant activities are found to be 289±14.6 AAE/mg, 468±22.7 GAE/mg. The observed antioxidant activities could be correlated with metabolites such as polyphenol, flavonoid, and terpenoid group of compounds identified in hexane fraction of E. repens by 4D GCXGC TOF-MS. Further ERH was evaluated for its protective properties against macromolecules such as DNA, protein and lipid damage. The extract showed protection against H2O2 induced DNA damage and inhibited AAPH induced protein oxidation and lipid peroxidation. Moreover, ERH administration to rats at 50 and 100mg/kg inhibited carrageenan-induced paw edema, and down-regulated cytokines such as COX-2, IL-6, and TNF-α and inhibited i-NOS mediated NO generation. E. repens also exhibited antioxidant effects by restoring SOD, catalase, GSH levels and inhibited lipid peroxidation in carrageenan challenged rats. Overall, the results suggest that E. repens may be useful in combating inflammation and oxidative stress.

Chemical composition of Ocimum sanctum by LC-ESI-MS/MS analysis and its protective effects against smoke induced lung and neuronal tissue damage in rats.

Smoke induced oxidative stress is known to cause various cancers and associated health problems including lung cancer. Herbal extracts have been reported as antioxidant supplements which attenuate free radical induced oxidative damage of tissues, among which Ocimum sanctum has been reported as the elixir of life due to its innumerable health benefits. In the present study, we investigated the protective effect of O. sanctum against cracker smoke induced lung and brain tissue damage. The results of the study demonstrate that O. sanctum regulates the hematological and serum biochemical parameters such as RBC, WBC, blood urea nitrogen and creatinine kinase. O. sanctum supplementation inhibited oxidative stress as analyzed by SOD, CAT enzyme levels and i-NOS, HSP-70 protein expression. O. sanctum administration also regulated neurotransmitter levels, such as serotonin, dopamine, and regulated acetylcholine esterase levels which play a vital role in neuronal function. Further O. sanctum treatment also preserved the morphology of lung and brain tissues of smoke stress induced rats as observed by histopathology and transmission electron microscope analysis. The biodistribution of O. sanctum was showed its accumulation in key tissues such as kidney, liver, lungs and heart. The LC-ESI-MS/MS analysis of O. sanctum showed the presence of polyphenols, flavonoids and fatty acids which might be responsible for the observed anti-stress effects.

Hypoxia induced cognitive impairment modulating activity of Cyperus rotundus.

Hypobaric hypoxia leads to decrease in cellular oxygen content which subsequently damages the hippocampus with an increase in brain oxidative stress and impairs the memory of the individual. In the present study, we have evaluated the cognitive impairment modulating activity of total oligomeric flavonoids fraction of Cyperus rotundus (TOF) in Sprague Dawley rats. The rats were trained for memory activity for a period of 7days followed by 7days exposure to 25,000ft. altitude and the spatial reference memory was evaluated. Behavioral analysis of the rats by Morris water maze experiment showed that TOF supplementation enhanced the spatial reference memory activity of the rats exposed to hypobaric hypoxia. The decrease in antioxidant status of the animals exposed to hypoxia was restored with TOF supplementation. The increase in ROS, lipid peroxidation products and protein carbonyls of the hippocampus was significantly decreased in animals with TOF administration. The histological assessment of the pyramidal cells of the hippocampus of hypoxia-exposed animals showed nuclear damage and TOF supplementation prevented nuclear damage. TOF administration suppressed hypoxia-induced increase in serotonin, dopamine, and norepinephrine. GABA and Ach levels were decreased by hypoxia which was prevented by TOF supplementation. The increase in GFAP, HIF-1α and VEGF expression in CA3 region of the hippocampus in hypoxia-exposed rats was decreased in TOF administered rats. Taken together, TOF extract ameliorates hypobaric hypoxia induced memory impairment and neurodegeneration in hippocampus through its anti-stress effects.

Growth Inhibition and Morphological Alteration of Fusarium sporotrichioides by Mentha piperita Essential Oil.

OBJECTIVE: The aim of this study is to determine the phytochemical composition, antifungal activity of Mentha piperita essential oil (MPE) against Fusarium sporotrichioides. METHODS: The phytochemical composition was conducted by gas chromatography mass spectrometry (GC MS) analysis and mycelia growth inhibition was determined by minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC), the morphological characterization was observed by scanning electron microscopy. Finally, the membrane permeability was determined by the release of extracellular constituents, pH, and total lipid content. RESULT: In GC MS analysis, 22 metabolites were identified such as menthol, l menthone, pulegone, piperitone, caryophyllene, menthol acetate, etc. The antifungal activity against targeted pathogen, with MIC and MFC 500 µg/mL and 1000 µg/mL, respectively. The MPE altered the morphology of F. sporotrichoides hyphae with the loss of cytoplasm content and contorted the mycelia. The increasing concentration of MPE showed increase in membrane permeability of F. sporotrichoides as evidenced by the release of extracellular constituents and pH with the disruption of cell membrane indicating decrease in lipid content of F. sporotrichoides. CONCLUSION: The observed results showed that MPE exhibited promising new antifungal agent against Fusarium sporotrichioides. SUMMARY: F. sporotrichioides, filamentous fungi contaminate to corn and corn--based productsF. sporotrichioides mainly responsible for the production of T-2 toxinPhytochemical composition was conducted by gas chromatography--mass spectrometry analysisMentha piperita essential oil (MPE) is commonly known as peppermintThe F. sporotrichioides growth was inhibited by MPE (minimum inhibitory concentration, minimum fungicidal concentration)Morphological observation by scanning electron microscope. Abbreviations Used: Cfu: Colony forming unit; DMSO: Dimethyl sulfoxide, °C: Degree celsius; F. Sporotrichoides: Fusarium sporotrichioides; EOs: Essential oils; M: Molar, g: Gram/gravity, mg: Milligram; µg: Microgram, ml: Milliliter; mm: Millimeter, min: Minutes; M. piperita: Mentha piperita, MIC: Minimum inhibitory concentration; MFC: Minimum fungicidal concentration; MAE: Mentha arvensis essential oil; Na2SO4: Sodium sulfate; pH: Potential Hydrogen; PDB: Potato Dextrose Broth; SEM: Scanning electron microscope.

Cytotoxic Effects of Ochratoxin A in Neuro-2a Cells: Role of Oxidative Stress Evidenced by N-acetylcysteine.

Ochratoxin-A (OTA), is toxic secondary metabolite and is found to be a source of vast range of toxic effects like hepatotoxicity, nephrotoxicity. However, the information available currently regarding neurotoxic effects exerted by OTA is scanty. Hence, the present study was aimed to evaluate the neurotoxic effects of OTA and the possible mechanisms of toxicity as well as the role of cytotoxic oxidative stress on neuronal (Neuro-2a) cell line was evaluated in vitro. Results of the MTT and LDH assay showed that, OTA induced dose-dependent cell death in Neuro-2a cells and EC50 value was determined as 500 nM. OTA induced high levels of reactive oxygen species (ROS) and elevated levels of malondialdehyde, also loss of mitochondrial membrane potential was observed in a dose depended manner. Effects of OTA on ROS induced chromosomal DNA damage was assessed by Comet assay and plasmid DNA damage assay in which increase in DNA damage was observed in Neuro-2a cells by increasing the OTA concentration. Further western blotting analysis of OTA treated Neuro-2a cells indicated elevated expression levels of c-Jun, JNK3 and cleaved caspase-3 leading to apoptotic cell death. Other hand realtime-Q-PCR analysis clearly indicates the suppressed expression of neuronal biomarker genes including AChE, BDNF, TH and NOS2. Further N-acetylcysteine (NAC) pretreatment to Neuro-2a cells followed by OTA treatment clearly evidenced that, the significant reversal of toxic effects exerted by OTA on Neuro-2a cells. In the present study, results illustrate that ROS a principle event in oxidative stress was elevated by OTA toxicity in Neuro-2a cells. However, further in vivo, animal studies are in need to conclude the present study reports and the use of NAC as a remedy for OTA induced neuronal stress.

Phosphatidylcholine, an edible carrier for nanoencapsulation of unstable thiamine.

Lipid nanoparticles have been used for carrying different therapeutic agents because of the advantage in improved absorption, bioavailability, targeted deliveries and reduction in the quantity of drugs required. The aim of the study was to prepare and characterize nanoliposomes containing thiamine hydrochloride and study their physicochemical stability as this vitamin is highly unstable. Phosphatidylcholine (PC) was used as an edible encapsulant. The average size of nanoliposomes was found to be 150 nm and zeta potential was -34 mV. The encapsulation efficiency was 97%. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) confirmed the size, spherical nature and smooth surface of the nanoliposomes. Differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) evidenced that the nanoliposomes were stable up to 300°C. The functional groups present were determined by Fourier transformed infrared spectroscopy (FTIR) and the presence of vitamin was confirmed in final formulation by biochemical analysis. The crystalline nature of thiamine was analyzed by X-ray diffraction studies. Storage studies indicated that the nanoliposomes were highly stable up to 3 months at different temperatures. Thus, phosphatidylcholine can be used as carrier vehicle of nutrients especially vitamins, as it can form stable nanoliposomes with 97% encapsulation efficiency.