Enzymatic Stress Markers Research Articles

Impact of microbial-based biopreparations on soil quality, plant health, and fruit chemistry in raspberry cultivation

Application of microbial-based biopreparations as a pre-harvest strategy offers a method to obtain sustainable agricultural practices and could be an important approach for advancing food science, promoting sustainability, and meeting global food market demands.The impact of a bacterial-fungal biopreparation mixture on soil-plant-microbe interactions, fruit chemical composition and yield of 7 raspberry clones was investigated by examining the structural and functional profiles of microbial communities within leaves, fruits, and soil. Biopreparation addition caused the enhancement of the microbiological utilization of specific compounds, such as d-mannitol, relevant in plant-pathogen interactions and overall plant health. The biopreparation treatment positively affected the nitrogen availability in soil (9–160%).The analysis of plant stress marker enzymes combined with the evaluation of fruit quality and chemical properties highlight changes inducted by the pre-harvest biopreparation application. Chemical analyses highlight biopreparations' role in soil and fruit quality improvement, promoting sustainable agriculture. This effect was dependent on tested clones, showing increase of soluble solid content in fruits, concentration of polyphenols or the sensory quality of the fruits. The results of the next-generation sequencing indicated increase in the effective number of bacterial species after biopreparation treatment. The network analysis showed stimulating effect of biopreparation on microbial communities by enhancing microbial interactions (increasing the number of network edges up to 260%) of and affecting the proportions of mutual relationships between both bacteria and fungi.These findings show the potential of microbial-based biopreparation in enhancing raspberry production whilst promoting sustainable practices and maintaining environmental homeostasis and giving inshght in holistic understanding of microbial-based approaches for advancing food science monitoring.

Palm Oil Mill Effluent Disposal and Its Utilization in Agricultural Soil

In the study, soil samples were impacted with 10%, 50% and 100% (v/w) of the palm oil mill effluents. Soil physicochemical properties, soil stress marker enzymes, soil bacteria populations were studied using standard methods. Analysis of the physicochemical properties of the Palm oil mill effluent (POME) showed that POME in the presence of the reference showed the following: pH, conductivity, BOD5, TDS, TSS, TS at 5.67 ±0.014a , 610±0.023c , 4.87±0.025b, 372.1±0.015c , 539.55±0.04a, 911.6±0.032b mg/ml, respectively dissolved mineral such as Magnessium (Mg), potassium (K), calcium (Ca) and phosphate (PO3) were recorded at 9.82±0.05b , 14.52±0.05a, 13.23±0.04c and 8.69±0.01bc mg/ml respectively. Total organic carbon and organic matter contents were recorded at 81.87±0.01a and 100.7±0.02c mg/ml respectively. Organismal proliferation increase as the percentage of the POME per gram of the soil increase from 10-50% (v/w) while a downturn in the organismal counts decreases significantly at 100% (v/w). There was a noticeable decrease in the coliform counts/g of the organisms as the incubation days increases from day 0-14. Enzyme activity relatively decreases as the contaminant concentrations increases from 0-100 v/w. Lipases activity was seen relatively low in all the soil at low concentrations of the effluents (0-50 v/w). At high concentrations of the effluents, there was a significant increase in the enzyme activity. POME can be utilized as sustainable source of organic agro fertilizer when quantified; however poor incentive by peasant farmers may lead to abysmal utilization of the waste water and its negative impact in the soil with aftermath of low soil fertility and poor agricultural productivity.

Profil Diferensiasi Leukosit dan Biomarker Stress Sapi Perah Laktasi dengan Pemberian Feed Supplement di Kelompok Ternak Tunas Mekar KSU Tandangsari

This research was conducted from April to June 2023 at the Tunas Mekar Cattle Group, KSU Tandangsari. As many as 16 Friesian Holstein (FH) dairy cows lactating 1-5, were used in this study to review the leukocyte differentiation and biomarker stress profiles and find out which treatment had the best effect on the leukocyte differentiation profile and biomarker stress. The study used a completely randomized design (CRD) consisting of 4 treatments and 4 replications. The treatments included P0 (control) consisting of 60% forage + 40% concentrate; P1 = 60% forage + 40% concentrate (97% concentrate + 3% protein by-pass); P2 = 60% forage + 40% concentrate (95% concentrate + 3% protein by-pass + 2% Ca-PUFA); and P3 = 60% forage + 40% concentrate (93% concentrate + 3% protein by-pass + 2% Ca-PUFA + 2% organic minerals). Based on the results of the study, it was shown that feeding supplementation had a significant effect (P<0.05) on the leukocyte differentiation profile and stress biomarkers of lactating dairy cattle in the Mekar Tunas Livestock Group, KSU Tandangsari. Lactation dairy cows treated with P3 = 60% forage + 40% concentrate (93% concentrate + 3% protein by-pass + 2% Ca-PUFA + 2% organic minerals) had lower mean leukocyte differentiation and biomarker stress than controls ( P<0.05). This can be interpreted that the provision of rations supplemented with feed supplements in the form of 3% protein by-pass + 2% Ca-PUFA + 2% organic minerals is able to reduce the level of heat stress as seen based on leukocyte levels and their differentiation and stress marker enzymes in the blood.

Ameliorative effects of <i>Cnestis ferruginea</i> root extract on carbon tetrachloride nephrotoxicity in albino Wistar rats

The objective of this research was to assess the potential protective properties of ethanol extract derived from Cnestis ferruginea against the toxic effects by carbon tetrachloride (CCl4) in the kidneys of rats. In this study, male albino Wistar rats were pre-treated with C. ferruginea at doses of 100 and 200 mg/kg body weight (b.w.) on a daily basis for a duration of 14 days. Subsequently, a single dosage of CCl4 was administered intraperitoneally on the 7th day of the experiment. The rats were euthanized 24 hours after they were subjected to treatments using the extract on the 15th day. Subsequently, serum and kidneys were obtained and utilized for the analysis of biochemical parameters. Rats grouped as the control which were administered CCl4 experienced kidney damage, as indicated by a statistically significant increase (P < 0.05) in the measured parameters (urea, creatinine, uric acid in serum and malondialdehyde in kidney tissue), and a significant reduction in the levels of oxidative stress marker enzymes (superoxide dismutase, catalase and glutathione peroxidase). It is worth noting that the administration of C. ferruginea to rats exposed to CCl4 resulted in a significant reversal of the aforementioned changes, bringing them closer to a state of normalcy. The result obtained from this study suggest that C. ferruginea possess therapeutic properties which could protect the kidney against renal deterioration and its associated dysfunction.

Curculigo pilosa rhizome Supplementation Reverses P-Hydroxyacetanilide-Induced Oxidative Stress and Lipid Profile Dysfunction in Rats

Exposure to drug toxicity results in stress and dysfunction of metabolizing enzymes that cause illnesses and diseases. Consumption of medicinal plant rhizomes is a regime for managing the complications. On the activities of some oxidative stress marker enzymes (catalase, glutathione, and superoxide dismutase), as well as the concentration of lipid profiles (cholesterol, triglycerides, and phospholipids) of p-hydroxyacetanilide (pPHA)-induced toxicity in rats, the effects of an aqueous extract of Curculigo pilosa (C. pilosa) rhizome were assessed. Forty rats were randomly grouped into eight groups (n = 5). The control group; aqueous extract of C. pilosa rhizome group; 750 mg/kg and 1000 mg/kg per body weight of pPHA group; the preventive groups (aqueous extract of C. pilosa rhizome and 750 mg/kg per body weight of pPHA; aqueous extract of C. pilosa rhizomes and 1000 mg/kg per body weight of pPHA); and the curative groups (750 mg/kg per body weight of pPHA and aqueous extract of C. pilosa rhizome; 1000 mg/kg body weight of pPHA and aqueous extract of C. pilosa rhizome). The oxidative stress marker enzymes and lipid profiles were analyzed spectrophotometrically in the serum, kidney, brain, and liver of the animals on the seventh and fourteenth days after the administrations. The results show that pPHA decreases the oxidative stress marker activities and the lipid profile concentrations in all the compartments, but the pre- and post-treatment with an aqueous extract of C. pilosa rhizome improved the activities of the stress marker enzymes and the lipid profiles dysfunction. The result suggests that an aqueous extract of C. pilosa rhizome has preventive and curative therapeutic potential for pPHA-induced toxicity.

Variations in antioxidant status in ischemic stroke: A case control study in Indian population

Objectives An imbalance between the levels of oxidants and antioxidants, a state of oxidative stress, leads to production of Reactive Oxygen Species. Overproduction of these species plays a crucial role in pathogenesis of ischemic stroke. In view of this, we aimed to investigate the markers of oxidative stress along with blood cellular components in ischemic stroke patients and healthy controls. Material and Methods Blood samples of 68 patients and 52 aged-matched controls were analyzed for oxidative stress markers such as glutathione peroxidase (GSHPx) and reduced glutathione (GSH) were assayed along with catalase and superoxide dismutase (SOD). Ferric-reducing antioxidant power (FRAP) of plasma of all samples was also evaluated along with complete blood count. Results In patients, the activities of the oxidative stress marker enzyme, GSHPx, was found to be significantly elevated (P ≤ 0.01), with significantly low levels of GSH (P ≤ 0.01), while the activities of catalase and SOD were found unaltered. The total antioxidant capacity of plasma measured as FRAP was also significantly higher (P ≤ 0.05). Red blood cells (RBCs) and eosinophils counts were found to be significantly lower (P ≤ 0.05 and P ≤ 0.01, respectively) while that of monocytes were significantly higher (P ≤ 0.01) in comparison with the controls. Conclusion Our results showed that the antioxidant status of ischemic stroke patients was better than the control subjects, which may have been due to the compensatory mechanism being activated due to oxidative stress.

Neuroinflammation, Energy and Sphingolipid Metabolism Biomarkers Are Revealed by Metabolic Modeling of Autistic Brains

Autism spectrum disorders (ASD) are a heterogeneous group of neurodevelopmental disorders generally characterized by repetitive behaviors and difficulties in communication and social behavior. Despite its heterogeneous nature, several metabolic dysregulations are prevalent in individuals with ASD. This work aims to understand ASD brain metabolism by constructing an ASD-specific prefrontal cortex genome-scale metabolic model (GEM) using transcriptomics data to decipher novel neuroinflammatory biomarkers. The healthy and ASD-specific models are compared via uniform sampling to identify ASD-exclusive metabolic features. Noticeably, the results of our simulations and those found in the literature are comparable, supporting the accuracy of our reconstructed ASD model. We identified that several oxidative stress, mitochondrial dysfunction, and inflammatory markers are elevated in ASD. While oxidative phosphorylation fluxes were similar for healthy and ASD-specific models, and the fluxes through the pathway were nearly undisturbed, the tricarboxylic acid (TCA) fluxes indicated disruptions in the pathway. Similarly, the secretions of mitochondrial dysfunction markers such as pyruvate are found to be higher, as well as the activities of oxidative stress marker enzymes like alanine and aspartate aminotransferases (ALT and AST) and glutathione-disulfide reductase (GSR). We also detected abnormalities in the sphingolipid metabolism, which has been implicated in many inflammatory and immune processes, but its relationship with ASD has not been thoroughly explored in the existing literature. We suggest that important sphingolipid metabolites, such as sphingosine-1-phosphate (S1P), ceramide, and glucosylceramide, may be promising biomarkers for the diagnosis of ASD and provide an opportunity for the adoption of early intervention for young children.

Defense Inducers Mediated Mitigation of Bacterial Canker in Tomato through Alteration in Oxidative Stress Markers.

The bacterial canker disease of tomato caused by Clavibacter michiganensis subsp. michiganensis (Cmm) has been reported to adversely affect the tomato cultivation in the NE hilly regions of India. Defense inducers such as salicylic acid (SA), isonicotinic acid (INA), benzothiadiazole (BTH) and lysozyme were used as prophylactic and curative sprays at different concentrations to test their efficacy in inducing resistance in tomato plants against Cmm under protected conditions. The induced resistance was studied through the alteration in the activities of oxidative stress marker enzymes (PAL, PO, PPO, TPC and PR-2 protein), hydrogen peroxide formation in leaf tissues and lignin accumulation in stem tissues, as well as through the reduction in disease severity under glasshouse conditions. The results of the present study revealed that the enzymatic activity, hydrogen peroxide formation and lignin production were significantly higher in the BTH (500 ppm)-treated leaves than in those observed in the control. The lowest disease incidence was recorded when BTH was applied as a prophylactic spray (27.88%) in comparison to being applied as a curative spray (53.62%), thereby suggesting that a defense inducer, BTH, shows antibacterial activity against Cmm, reduces disease incidence severity and induces defense responses in the tomato plant.

Effects of environmentally relevant concentrations of microplastics on amphipods

Lack of microplastics (MP) toxicity studies involving environmentally relevant concentrations and exposure times is concerning. Here we analyzed the potential adverse effects of low density polyethylene (LDPE) MP at environmentally relevant concentration in sub-chronic exposure to two amphipods Gmelinoides fasciatus and Gammarus lacustris, species that naturally compete with each other for their habitats. 14-day exposure to 2 μg/L (8 particles/L corresponding to low exposure) and 2 mg/L (∼8400 particles/L, corresponding to high exposure) of 53–100 μm LDPE MP were used to assess ingestion and egestion of MP, evaluate its effects on amphipod mortality, swimming ability and oxidative stress level. Both amphipod species were effectively ingesting and egesting LDPE MP. On the average, 0.8 and 2.5 MP particles were identified in the intestines of each amphipod exposed to 2 μg/L and 2 mg/L LDPE MP, respectively. Therefore, intestinal MP after 14-day exposure did not fully reflect the differences in LDPE MP exposure concentrations. Increased mortality of both amphipods was observed at 2 mg/L LDPE MP and in case of G. lacustris also at 2 μg/L exposure. The effect of LDPE on swimming activity was observed only in case of G. fasciatus. Oxidative stress marker enzymes SOD, GPx and reduced glutathione GSH varied according to amphipod species and LDPE MP concentration. In general G. lacustris was more sensitive towards LDPE MP induced oxidative stress. Overall, the results suggested that in MP polluted environments, G. lacustris may lose its already naturally low competitiveness and become overcompeted by other more resistant species. The fact that in the sub-chronic foodborne exposure to environmentally relevant and higher LDPE MP concentrations all the observed toxicological endpoints were affected refers to the potential of MP to affect and disrupt aquatic communities in the longer perspective.

Risk Assessment of Heavy Metal Toxicity and Stress Marker Enzyme in Waste Water from Municipal Dam Site around Enugu Metropolis, Enugu State

Impact of waste water toxicity from collection ditches within Enugu metropolis, Enugu state was assessed in the present study. Risk impact of heavy metal recalcitrant was also determined in the present study as the waste water from the ditch were used by subsistence farmers around for irrigation purposes.  Physicochemical analysis of the soil irrigation with the domestic waste water was carried and revealed the following: pH 6.65, conductivity 792 respectively. Dissolved mineral contents were found in the following order: Cl>Ca>Fe>SO4>Mg>Cu>K>Pb>PO3 while heavy metals of Hg, As and Cd were found at below detectable limit range (BDL). Total organic carbon (TOC) and total organic matter (TOM) contents were found at 179.93 and 221 mg/kg in the various treatments. These varied significantly from the control treatment. Physicochemical properties of the domestic waste water were equally analyzed.  Soil enzymes showed the activity of the lipase, urease, peroxidase and catalase with an increased of lipase, urease and peroxidase after the irrigation with OD of 0.610, 0.677 and 0.712 respectively catalase activity was reduced downwardly. Health risks to human through utilization of the waste water by human were assessed by: estimated dietary intake (EDI), estimated weekly intake (EWI) and Target hazard quotient (THQ). EDI for the heavy metals of Pb, Cu and Fe were 3.98 x 10-3, 1.22 x 10-2 and 1.2 x 10-2 mg/g, respectively estimated while EDI of 1.36 x 10-3, 8.33 x 10-3 and 9.52 x 10-3 mg/g were obtained for Pb, Cu and Fe from the control experiments. EWI for heavy metals of Pb, Cu and Fe were 9.52 x 10-3, 583 x 10-2 and 6.7 x 10-2 mg/g, respectively. The results showed that the THQ for Fe and Cu were less than 1 while that of Pb was greater than 1. THQ were less than 1in all the analysed metals from the control experiment. The findings from this study show that there is a considerable health risks connected with utilization of waste water from our study locale and these waste water bioaccumulate heavy metal levels which are potentially poisonous to humans who consume them.

Protective effects of N(1)-2,4-dihydroxybenzylidene-N(4)-2-hydroxybenzylidene-S-methyl-thiosemicarbazidato-oxovanadium (IV) on oxidative brain injury in streptozotocin-induced diabetic rats.

Diabetes is usually accompanied by increased production of free radicals or impaired antioxidant defenses. The brain is a target tissue of the oxidative attacks caused by diabetes, and there are observed changes in the biochemical parameters of this tissue in the hyperglycemic state. In this study, we aimed to show the effect of N(1)-2,4-dihydroxybenzylidene-N(4)-2-hydroxybenzylidene-S-methyl-thiosemicarbazidato-oxovanadium (IV) (VOL) compound on diabetic damaged brain tissue, induced by streptozotocin (STZ) on 3.0-3.5-month-old male rats. Single dose of STZ at 65 mg/kg was used to make rats diabetic. Four groups were created randomly. Group (i): control (intact) animals; Group (ii): VOL given control animals; Group (iii): STZ-induced diabetic animals; and Group (iv): orally VOL administered STZ-induced diabetic rats. VOL (0.2 mM/kg/day) administration to control and diabetic animals was performed for a period of 12 days. At the end of day 12, the brain tissues were taken and homogenized. The clear supernatants were used for the determination of glutathione (GSH), lipid peroxidation (LPO), nonenzymatic glycosylation (NEG), and protein levels. Alanine and aspartate transaminases and acetylcholinesterase (AChE), myeloperoxidase (MPO), xanthine oxidase (XO), and oxidative stress marker enzymes activities were also estimated from the homogenates. According to the obtained results, there is found significant elevation of MDA and NEG levels and activities of transaminases, MPO and XO; whereas the GSH content and the activities of AChE and antioxidant enzymes were strongly decreased in the STZ-induced diabetic brain tissues in comparison to control group animals. Twelve days of administration of VOL complex to the diabetic animals reversed all biochemical parameters significantly in diabetic brain tissues. Our findings suggest that the VOL complex may be an ideal candidate to be used as an anti diabetic agent to improve oxidative injury and protect the brain tissue against damage caused by diabetes. This healing effect of the VOL complex may be due to its antioxidant activity and the insulin-mimetic effects of vanadium.

Protective Role of Vitamin C Against Biochemical Enzymes Alterations Induced by Exposure of Allethrin Based Mosquito Coil Smoke on Cerebellum of Male Albino RatsAn Experimental Study

Introduction: Among synthetic pyrethroids allethrin is widely used as active ingredient in mosquito coils. Many experimental studies have proved that pyrethroids cause neurotoxicity and result into derangements of biochemical enzymes including oxidative stress markers and antioxidant levels in brain. Aim: To study alterations in biochemical markers (oxidative and antioxidant enzymes) in cerebellum of male albino Wistar rats induced by the exposure of allethrin based mosquito coil smoke along with protective role of withdrawal and exogenously administered vitamin C on these markers. Materials and Methods: This experimental study was conducted in the Department of Anatomy, King George's Medical University, Lucknow, Uttar Pradesh, India, during October 2014-September 2015. Total 42 male albino Wistar rats were divided into four groups. The first group contained 12 rats was used as control, group II with 12 rats was given 8 hours daily exposure of allethrin based mosquito coil smoke via whole body inhalation for 6 days in a week for a total of 12 weeks. Group III contained eight rats and was given exposure same as group II and were further kept for 8 weeks to see the withdrawal effect. Group IV with 10 rats was given same exposure as group II, along with oral administration of 20 mg/kg body weight vitamin C. After 12 weeks of exposure cerebellum was dissected and then sent for biochemical enzyme estimation. Rats in group III were dissected after 8 weeks post 12 weeks of exposure. Comparisons were made between groups by using one way Analysis of Variance (ANOVA). Results: The exposure Group II showed significant rise (p<0.001) in level of oxidative stress marker enzyme Malondialdehyde (MDA) and significant (p<0.001) reduction in activities of antioxidants Glutathione Peroxidase (GPO), reduced Glutathione (GSH), Superoxide Dismutase (SOD) and Catalase (CAT) in comparison to Group I, III and IV. In vitamin C administered group IV the above parameters were found to be less affected. Intergroup comparison of oxidative stress markers level showed significant intergroup difference between Group I, II, group II, III and group II, IV. Conclusion: Allethrin based mosquito coil smoke causes oxidative stress in cerebellum of albino Wistar rats and vitamin C has ameliorative effect on biochemical alterations induced by sub-chronic whole body exposure of mosquito coil smoke.

ANALYSIS OF OXIDATIVE STRESS MARKERS IN CHRONIC CONSUMPTION OF MONOSODIUM GLUTAMATE ON LIVER OF WISTAR ALBINO RATS

Objective: The study was intended to explore whether Monosodium glutamate (MSG) induces oxidative stress on the liver of Wistar albino rats when fed chronically at three different doses, namely, low, mid, and high doses identical to human consumption doses in growing countries. Methods: The acclimatized Wistar albino rats (n=24) were randomly selected and grouped into four groups, namely Control, Low dose MSG (180 mg kg), Mid dose MSG (360 mg/kg), and High dose MSG (720 mg/kg). The animals were orally administered MSG for 120 days. After completion of the experimental period (120 days), euthanized animal liver was homogenized to investigate the oxidative stress marker enzymes such as Superoxide Dismutase (SOD), Glutathione Peroxidase (GPx), Catalase (CAT), and Myeloperoxidase (MPO). Results: The MPO showed a significant increase (p<0.05) in liver homogenate of all MSG induced groups when compared to control group. The SOD, CAT, and GPx activity deteriorated (p<0.05) in monosodium induced groups contrasting to the control group. Conclusion: The effects of MSG on oxidative stress markers on liver homogenate in the current study exhibited erratic abnormal changes in oxidative stress markers of monosodium induced groups which contemplate the harmful effects of MSG consumed chronically. The further studies should confirm the genetic basis of oxidative stress damage and transform the safety regulations of MSG consumption throughout the world.

Radio-protective efficacy of Gymnema sylvestre on Pangasius sutchi against gamma (60Co) irradiation

ABSTRACT Purpose Freshwater fish Pangasius sutchi was used in this study as a vertebrate model. We evaluated the induction of certain antioxidant enzymes in various vital organs. The radioprotective efficacy of Gymnema sylvestre leaves extract (GS) [25mg/kg Body Weight (B.W)] and its bioactive compound Gymnemagenin (GG) [0.3mg/kg B.W] was compared with Amifostine (Ami), the only radioprotector clinically approved by the US-FDA [Ami- 83.3mg/kg B.W] against different doses of gamma radiation – 60Co (Lethal Dose: LD30-9.2Gy, LD50-10.2Gy and LD70-11.4Gy). Materials and Methods This study was done via stress marker enzymes, cell cycle analysis (CCA) and DNA damage assay prediction with molecular docking, which are reported here for the first time. The results indicate an elevated LPO level and decreased level of CAT, SOD and GSH due to oxidative stress initiation by 60Co Ionizing Radiation (IR) on 4th day and slightly reduced on 32nd day while the reverse observed when the fishes were pre-treated with Ami, GS and GG. Similarly, CCA and dead/live cells counts were conducted with pre-treatment of Ami, GS and GG against 60Co IR dose (LD50-10.2Gy). Results In CCA, G0/G1 phase was observed to be the highest in Ami and lowest in GG, against 60Co IR doses 10.2Gy which was 51.76 ± 7.55. The dead cells range observed in pre-treated group of Ami, GS and GG was lowest in Ami and highest in GG and live cells (highest in Ami and lowest in GG) as compared to 60Co IR group (86.43 ± 3.42 and 8.77 ± 5.95). Thus, antioxidant profile improvement by oxidative stress reduction and gradual progression of different phases of cell cycle except the apoptotic phase along with the live cells counts indicates that the radio-protective efficacy of GS is similar to Ami. Conclusion Predictive assessment was carried out by docking of Ami, various components of GS with p53, NF-κβ cells and Rad51 proteins structures responsible for CCA, apoptosis and repair mechanism. These structural proteins were docked with other structural proteins like USP7, TNF-α and partner and localizer of BRCA2 associated (PALB2/BRCA2) complex which made us perform these systemic efforts to find the functional activity of these known radio-protectants.

Hepatoprotective Property of Phyllanthus amarus Alkaloid-Rich Fraction with Respect to Oxidative Stress Marker Enzymes

A disruption in the equilibrium between the generation of reactive oxygen species and antioxidant defense enzymes is referred to as oxidative stress. In the present study, we planned to identify the hepatoprotective effect of Phyllanthus amarus alkaloid rich fraction in wistar strain albino male rats. The hepatic damage was induced by the D-galactosamine and ameliorative effect was tested with alkaloid rich fraction of P. amarus by measuring oxidative stress markers such as G6PDH, LDH, SDH, MDH and GDH in the liver tissue. Activity levels of G6PDH, SDH, MDH and GDH were significantly decreased in D-galactosamine induced hepatitis rats when compare to normal control rat group, while their activities were significantly increased in hepatitis rat group that supplemented with alkaloid rich fraction of P. amarus. In contrast, LDH enzyme activity of liver was significantly increased in the hepatitis rat group when compare to normal control rats, while its activity was significantly decreased in hepatitis rats treated with alkaloid fraction. In conclusion, it is very clear that alkaloid fraction of P. amarus has hepatoprotective property with respect of decreasing oxidative stress by regulating oxidative stress marker enzymes. The isolation and identification of specific alkaloid compounds with hepatoprotective properties and anti-oxidative stress will require much further research.

Anticancer Effect of Troxerutin in Human Non-Small-Cell Lung Cancer Cell A549 and Inhibition of Tumor Formation in BALB/c Nude Mice.

This study is intended to explore the anticancer, antiproliferative, and chemopreventive action of troxerutin (TX) in human non-small-cell lung cancer cell (A549) using BALB/c nude mice. 2 × 106 A549 cells were subcutaneously injected into mice, along with 10 μM and 20 μM/kg body weight of TX orally for 19 days. On the last day, tumor weight and volume were assessed. Stress marker enzymes such as Aryl hydrocarbon hydroxylase (AHH), lactate dehydrogenase (LDH), 5'Nucleotidase (5'ND), and γ-glutamyltranspeptidase (γ-GT) were estimated in the lung tissues. Cytotoxicity of TX was assessed using MTT assay. Expression of carcinoembryonic antigen (CEA) and inflammatory cytokines were also analyzed. Histopathological examination of tissue sections and immunohistochemical examination of proliferating cell nuclear antigen (PCNA) were also performed. mRNA expression of p53, p21, cyclin D1, P13k, Akt, and mTOR were analyzed using RT-PCR. TX administered orally in a dose-dependent manner markedly reverted the level of stress marker enzymes to a significant extent. TX also exhibited significant protection against lung cancer cells, as evidenced by cytotoxicity assay and histopathological studies. It was also found to reduce the expression of PCNA, cyclin D1, P13k, Akt, and mTOR, but increase the expression of p53 and p21. TX has also been shown to reduce cancer cell inflammation, as was evidenced by reduced expression of inflammatory cytokines. Thus TX could be used as an effective chemopreventive and anticancer agent in treating cancer.

Experimental Evaluation of Prophylactic and Therapeutic Antioxidant Potential of Trimetazidine in Carbon Tetrachloride Induced Oxidative Stress in Rats

Introduction: Exposure to various drugs and chemicals lead to oxidative stress. Carbon Tetrachloride (CCl4) produces rise in oxidative stress leading to hepatic damage. The drug Trimetazidine (TMZ) shows hepatoprotective activity but its mechanism is not known. The present study would help in establishing antioxidant activity of TMZ as probable mechanism. Aim: To evaluate the antioxidant potential of TMZ in CCl4 induced oxidative stress when given prophylactically/therapeutically in rats. Materials and Methods: An experimental animal study was conducted on 80 adult Wistar rats of either sex (weight-150 to 200 gm) from March 2010 to December 2010 in Bharati Vidyapeeth Medical College, Pune, Maharashtra, India. Randomly, all animals were grouped into 10 equal groups. Group i was normal control (received only water). To induce oxidative stress CCl4 (0.5 mL/kg/d i.p.) was given to all the animals of Group ii to Group x for seven days. The TMZ was given in two doses, TMZ1 (5 mg/kg orally for Group iii and vii) and TMZ2 (10 mg/kg orally for Group iv and viii). Positive standard control (Group v and Group ix) received Liv.52 (1 mL/kg orally). Group vi and Group x received combination of TMZ1 (5 mg/kg orally)+Liv.52 (1 mL/kg orally). Drug treatment was given to animals in group iii, iv, v and vi for 1-14 days (preventive group) and in group vii, viii, ix and x from day 8 to day 14 (therapeutic group). On 15th day, rats were sacrificed and dissected for collection of liver. Part of the livers was homogenised to assess oxidative stress marker enzymes Malondialdehyde (MDA), Superoxide Dismutase (SOD) spectrophotometrically. Statistical analysis was done with one- way Analysis of Variance (ANOVA) followed by post-hoc analysis (Dunnett’s test) using GraphPad Prism 5.0 software. Results: Trimetazidine (5 mg/kg and 10 mg/kg) significantly reduced MDA levels and increased SOD levels when compared with CCl4 treated group suggested antioxidant activity. Combined administration of Liv.52 and TMZ1 also reduced oxidative stress and increased antioxidant activity. Conclusion: Results of the present study suggested that increased oxidative stress was significantly attenuated by drug TMZ in dose dependant manner when compared with the CCl4 group. The antioxidant potential of prophylactic and therapeutic administration of TMZ was comparable. The increased antioxidant effect by Liv.52+TMZ1 combination was only due to the additive antioxidant effects of Liv.52 and TMZ or any other mechanism was involved, needs to be further evaluated.

Involvement of steroid and antioxidant pathways in spleen-mediated immunity in migratory birds

The molecular underpinnings of the spleen-mediated immune functions during the period of heightened energetic needs in the year are not known in avian migrants. We investigated this, in Palearctic-Indian migratory male redheaded buntings, which exhibited vernal (spring) premigratory / early testicular maturation states under artificial long days. This was evidenced by increased dio2 and decreased dio3 mRNA expression in the hypothalamus, elevated levels of circulating corticosterone and testosterone, and enlarged testes in long-day-photostimulated birds, as compared to unstimulated controls under short days. The concomitant decrease in both mass and volume of the spleen, and increase in the heterophil/ lymphocyte ratio suggested the parallel innate immunity effects in photostimulated buntings. Importantly, we found increased mRNA expression of genes coding for the cytokines (il15 and il34), steroid receptors (nr3c2) and oxidative stress marker enzymes (gpx1 and sod1) in the spleen, suggesting the activation of both immune and antioxidant molecular pathways during the early photostimulated state. However, the splenic expressions of il1β, il6, tgfβ, ar and nos2 genes were not significantly different between long-day stimulated and short-day unstimulated birds. The negative correlation of plasma corticosterone levels with spleen mass further indicated a role of corticosterone in the modulation of the spleen function, probably via nr3c2 gene encoded mineralocorticoid receptors. These results suggest the activation of the spleen-mediated innate immunity in anticipation of the heightened energetic stress state of the photostimulated spring migratory/breeding period in migratory songbirds.

Effect of citrate stabilized gold nanoparticle on the biochemical and histological alterations of liver in an experimental animal model

Although the nanoparticles (NP) are seemed to have the immense potentiality for using in different biomedical fields the possible toxic health effects of these NPs associated with human exposure is a serious concern. In general, the toxicity of gold nanoparticles (AuNPs) depends on their physical dimensions and surface chemistry. In the present study, size-specific AuNPs were used to test its potential harmful effect in the biochemical and histological alterations of liver tissue in experimental animals (swiss albino mice). Animals were exposed to the AuNPs for seven days. Oxidative stress in whole tissue homogenates and in mitochondrial extracts and histological studies were studied in the liver. Significant deterioration in glutathione reductase (GR), malondialdehyde (MDA), and total thiol content were observed in the case of whole liver tissue homogenates; on the other hand, in case of whole-brain tissue homogenates, deterioration in glutathione S-transferase (GST), total thiol and MDA was observed. Almost all the lipid peroxidation product and oxidative stress marker enzymes were affected in the hepatic system were observed in AuNP treated animals. Anatomical differences in liver tissue were also observed in the AuNP treated animals. Minute loss of hepatic cords, occurrence of swallowing and atrophic cells is the major detoriation observed in AuNP treated animals. Surface negativity and the small size of the AuNPs may be assumed to affect the biochemical and cellular architecture of the organs. The further molecular level analysis will reveal the usability potential of AuNP in the body system for different purposes.

Essential oil yield estimation by Gas chromatography-mass spectrometry (GC-MS) after Methyl jasmonate (MeJA) elicitation in in vitro cultivated tissues of Coriandrum sativum L.

Coriandrum sativum is an important spice plant known for its unique fragrance. Coriander oil is also one of the major essential oils in world global market. The oil yield varies with different coriander varieties; and the content and quality of oil is governed by several factors. In recent times, a variety of technologies have been exploited to improve phyto-compounds including essential oils. In this present study, Methyl jasmonate (MeJA) was amended in medium and the yield of essential oil was measured and compared in different cultivating tissues. The cultured tissues were nonembryogenic callus and embryogenic tissues (induction, proliferation and maturation stages of embryos). MeJA acts as a signaling molecule in accumulating secondary metabolites. Four different MeJA treatments i.e. T1 = 50, T2 = 100, T3 = 150 and T4 = 200μM, along with a control (T0) were used and the yield of coriander essential oil was estimated in different in vitro cultivating tissues by using Gas chromatography-mass spectrometry (GC-MS). The addition of MEJA enriched essential oil yield, maximum oil being in maturation stage of embryos at T3 (150μM). Other added treatments also had varied stimulatory role. The addition of MeJA induced stress as the stress marker enzymes like superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) content were high compared to non treated tissue (T0). In T4, the CAT activity was maximum i.e. 5.83 and 6.28mg-1 protein min-1 in Co-1 and RS respectively in matured somatic embryos. The SOD activity was also high at maturation stage of embryos at T4 (5.3mg-1 protein min-1 in RS). The APX activity on the other, was high (3.32mg-1 protein min-1) in induction stage of embryogenesis at T3. The comparative biochemical (sugar, protein and proline) analyses of tissues were performed and presented that had high and low essential oil. MeJA induced stress may help in accumulating essential oils in C. sativum.