Lipid Peroxidase Research Articles

Transient application of foliar urea with N-(n-Butyl) thiophosphoric triamide on N metabolism of pineapple under controlled condition

Traditional fertilization and the loss of Nitrogen (N) is a common phenomenon in agriculture. Application of urea with urease inhibitor is an alteration measure of urea hydrolysis and N metabolism. Hence, the objective of this study was to determine the short-term foliar application effect of N-(n‑butyl) thiophosphoric triamide (NBPT) on N metabolism, physiological and biochemical changes in the pineapple plant. At 4 month after planting (MAP), 1% urea (Urea) and 1% urea with NBPT (NLU) were applied. Data on urease enzyme activity, internal urea content, inorganic N [ammonium (NH4+) and nitrate (NO3−)], amino acids, total soluble protein, NBPT content, lipid peroxidase (MDA) activity, and electrolyte leakage were recorded for 10 days after treatment (DAT). The results indicated that the NBPT was taken up by the plant, thereby a clear inhibition of the urease activity was observed in pineapple leaf. A higher amount of urea content and a lower amount of NH4+ content in the leaf were the evidence of hydrolysis inhibition by NBPT. Consequently, the contents of amino acid and protein were diminished upon NBPT treatment which demonstrated a change in N metabolism. In addition, amino acid composition demonstrated that amide forms of amino acids were the major components of the decreased amino acid pool. In conclusion, lower MDA activity and electrolyte leakage were also observed under NBPT treated plants indicate that NBPT could release abiotic stress to some extent by protecting the cell wall from damage.

Clinico-pathological investigation of inflammatory mammary carcinomas in bitches

A study was conducted to elucidate the altered clinico-pathological parameters in diagnosis and prognosis of inflammatory mammary carcinomas in bitches. Blood and serum samples were collected from 30 each normal and tumor bearing bitches for haemato-biochemical analysis. A significant (P<0.05) reduction of total erythrocyte count and highly significant (P<0.01) reduction of packed cell volume were noticed in tumor affected bitches. A significant (P<0.05) elevation of mean corpuscular haemoglobin and highly significant (P<0.01) elevation of mean corpuscular haemoglobin concentration and total leucocyte count were observed in mammary tumor affected dogs. A significant (P<0.05) increase in total cholesterol and highly significant (P<0.01) increase in alanine transaminase, aspartate transaminase, alkaline phosphatase and potassium values were noticed in tumor bearing bitches. Highly significant (P<0.01) reduction in albumin and significant (P<0.05) reduction in calcium were also noticed. An elevation of superoxide dismutase, reduced glutathione, glutathione peroxidase and lipid peroxidase values whereas reduction of catalase value were noticed in the tumor tissues. Histopathologically, the tumors were classified as benign (2) and malignant (28) tumors with infiltration of inflammatory cells in 18 cases. The haemato-biochemical alterations due to antioxidants and lipid peroxidase were consistent in the diagnosis and management of mammary tumors in bitches.

Effect of food colorant allura red on hematological, biochemical & antioxidant parameters of Swiss albino male mice

Colour additives are added in food to enhance the appearance of foodstuff. Today synthetic food colour are usually preferred than natural colour due to intense coloration and low cost. The aim of the present study is to observe the changes in the hematological, biochemical, and antioxidant parameters in Swiss albino male mice after 30, 45 and 60 days of treatment respectively with allura red. The experimental plan included 4 groups and in each group 6 mice were taken for statistically valid results. Group I served as control (received distilled water only), group II received allura red (AR) treatment (172.2 mg/kg body weight) for 30 days. Group III received AR treatment (172.2 mg/kg body weight) for 45 days and group IV received AR treatment (172.2 mg/kg body weight) for 60 days. The results revealed a noticeable decrease in white blood cell (WBC) count, red blood cell (RBC) count, haemoglobin (Hb), haematocrit percentage (Hct) and platelets after 60 days of treatment. A significant decrease was also observed in CAT (catalase), SOD (superoxide dismutase) and GSH (glutathione) values in the treatment groups. However, marked increase was observed in mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), Serum alkaline phosphatase (ALP), Serum total protein, Albumin, GPx (glutathione peroxidase) and LPO (lipid peroxidase) values. The variations in the different parameters were highly significant after 60 days of treatment. In conclusion, it was clear that the administration of allura red in mice caused significant variations in haematological, biochemical and antioxidant parameters indicating allura red toxicity at the given dose level.

Review: Bioavailability and Activity Prediction of Bioactive Compounds of Red Fruit (Pandanus conoideus Lam.) and Pandan Grape (Sararanga sinousa Hemsley) by in silico Method

Red Fruit (RF) and Pandan grape (PG) contained bioactive compounds that good for health. The structure influenced the availability and activity, including the interaction with another in the body. It needs time and financial consumption, whereas bioinformation technology offers the solution. The research aims to predict the availability and activity of RF and PG compounds by in silico method based on the SAR. Fourteen RF and PF phenolics were evaluated for availability, antioxidant, and antiglycation activity during binding to HSA. The flavonoids showed Pa&gt;0,7 for antioxidant in general, free radical scavenging, and lipid peroxidase inhibitor activity. The availability was shown by the ligand’s capability to bind to HSA. The order of affinity energy from the largest to the smallest was RF flavonoid &gt; PG anthocyanin &gt; GP phenolic acids. Taxifolin and Quercetin still had antioxidant activity during binding because there are free hydroxyl groups. PG chlorogenic acid and RF flavonoids play as an anti-diabetic through antioxidant mechanism and HSA glycation disturbance. Based on this review, in silico method is effective as an analysis tool for the activity and mechanism prediction of food bioactive compounds.

Stress Biomarkers of Metal Pollutants of Milkfish (Chanos chanos) in Estuarine Systems of North-East Coast of Bay of Bengal -A Case Study

Milkfish were sampled during premonsoon (2015) from the Hooghly River mouth Eco-region (HRME) and Saptamukhi River/Creek mouth Eco-region (SRME) on the Northeast of Bay of Bengal. Metal accumulation and its impact on the antioxidant systems of different tissues of the fish were studied. Study showed Zn &gt; Cu &gt; Pb &gt; Fe &gt; Mn &gt; Cd &gt; Ni in the gills, muscles, and liver. Metal concentration in the liver and gills was found to be quite high compared to the muscle. Active responses were recorded in cases of lipid peroxidase, superoxide dismutase, catalase, glutathione peroxidase in gill, liver and muscle. Reduced glutathione and glutathione-S-transferase activities could be attributed to metal induced oxidative stress. Alteration in the antioxidant enzymes, glutathione system and induction of lipid peroxidation underlined the impact of metals contaminants in the fish indicated to the stress experienced by a heavily consumed piscine species such as milkfish.

Acetyl-L-Carnitine protects against LPS induced depression via PPAR-γ induced inhibition of NF-κB/NLRP3 pathway

IntroductionMajor depressive disorder (MDD) is a debilitating human health status characterized by mood swings and high suicidal attempts. Several studies have reported the role of neuroinflammation in MMD, yet the efficacy of natural drug substances on neuroinflammation-associated depression needs to be further investigated. The present study demonstrated the neuroprotective effects of Acetyl-L- carnitine (ALC) alone or in combination with caffeic acid phenethyl ester (CAPE) on lipopolysaccharide (LPS) induced neuro-inflammation, depression, and anxiety-like behavior.Material and methodsMale Sprague Dawley (SD) rats were used to explore the relative effects of ALC and the mechanistic interplay of the peroxisome proliferator-activated receptors (PPARγ) in depression. Lipopolysaccharide (LPS) was administered to induce depression and anxiety-like symptoms such as a decreased grooming tendency, diminished locomotive activity, and increased immobility period.ResultsWe found marked neuronal alterations in the cortex and hippocampus of LPS intoxicated animals associated with higher inflammatory cytokines expression cyclooxygenase (COX2), tumor necrotic factor-alpha (TNF-α). These detrimental effects exacerbate oxidative stress as documented by a compromised antioxidant system due to high lipid peroxidase (LPO). ALC significantly reverted these changes by positively modulating the PPARγ dependent downstream antioxidant and anti-inflammatory pathways such as NOD and pyrin domain-containing protein 3 (NLRP3) linked nuclear factor kappa B (NF-κB) phosphorylation. Moreover, co-administering NF-κB inhibitor caffeic acid phenethyl ester (CAPE) with ALC also increased PPARγ expression significantly and decreased NF-ᴋB and NLRP3 inflammasome.ConclusionsThese findings indicate that ALC could be a possible depression supplement. The effects are partly mediated by inhibiting neuroinflammation and NLRP3 inflammasome coupled to PPARγ upregulations.

Effects of boric acid on ovarian tissue damage caused by experimental ischemia/reperfusion

ABSTRACT We investigated the role of boric acid (BA) in ovarian tissue damage caused by ischemia/reperfusion (I/R) in the rat. We established four groups of seven adult female rats: untreated control; ovarian I/R; 15 mg/kg BA; I/R + 15 mg/kg BA. Ovarian levels of lipid peroxidase (LPO), myeloperoxidase (MPO), glutathione (GSH), catalase (CAT), superoxide dismutase (SOD) and interleukin-6 (IL-6) were measured biochemically. Ovaries were evaluated for histopathology and investigated using immunohistochemistry. We detected greater LPO activity and less CAT, SOD and GSH levels in the I/R groups compared to the control group. LPO and MPO levels in ovarian samples for the I/R goup were increased significantly compared to untreated controls. The levels of LPO and MPO in ovarian tissue for the I/R + BA group were not significantly different from controls. SOD and GSH activity in ovarian tissue was increased significantly compared to controls. We found degenerated parenchymal cells, hemorrhage, necrotic parenchyma cells and congestion in the I/R groups. Expression of 8-OHdG was greater in the I/R group than in controls. Although immunostained cells were dense in the ovarian tissue in the I/R group, the number of immunostained cells was decreased by application of BA to the I/R group. BA exhibits a protective effect on ovarian tissue against I/R damage in the rat.

Plant-Derived Smoke Ameliorates Salt Stress in Wheat by Enhancing Expressions of Stress-Responsive Genes and Antioxidant Enzymatic Activity

Abiotic stresses are the biggest threat to the increasing population worldwide. Salt stress is one of the most significant abiotic stresses, affecting 20% of the crop production around the world. Plant-derived smoke (PDS) has been reported as a biologically active plant product in stimulating seed germination, seedling growth and physiological characteristics of crops under abiotic stress conditions. Nevertheless, studies showing how PDS alleviates salt stress are largely unknown. Here, we report the molecular mechanism of how PDS could alleviate salt stress in wheat. Initially, PDS at 2000 ppm enhanced seed germination, root/shoot length and seedling fresh weight. However, PDS at 1000 and 500 ppm did not show any significant effect. Salt stress at 150 and 200 mM significantly reduced seed germination rate, root/shoot length and fresh weight of the wheat seedlings. Interestingly, PDS supplementation at 2000 ppm concentration was sufficient to restore seed germination under salt stress condition. Moreover, PDS improved root/shoot length and seedling biomass under 150 and 200 mM salt stress, suggesting that PDS is a potent plant product, capable of abiotic stress alleviation in crops. In comparison to the control, PDS-treated seedlings displayed increased activity of major antioxidative enzymes such as superoxide dismutase, peroxidase and ascorbate peroxidase under salt stress, resulting in reduced levels of hydrogen peroxide and lipid peroxidase, showing that PDS can possibly help in salt stress amelioration by regulating redox homeostasis. Importantly, salt stress altered the expression of germination marker genes, such as TaSAM, TaPHY, TaBGU (germination positive effectors), TaLEA and TaGARS34 (germination negative effectors), suggesting the potential role of PDS in the germination pathway under salt stress. Further, PDS modulated the transcript levels of several salt stress stress-responsive genes, including TaSOS4, TaBADH and TaHKT2. In conclusion, this study provides a molecular and physiological basis for elucidating the mechanism of how PDS functions in stress induction in wheat, as well as demonstrates the importance of PDS in agricultural practices, laying the groundwork for future research into the role of PDS in the amelioration of abiotic stresses in various plants.

Enhanced Flavonoid Accumulation Reduces Combined Salt and Heat Stress Through Regulation of Transcriptional and Hormonal Mechanisms.

Abiotic stresses, such as salt and heat stress, coexist in some regions of the world and can have a significant impact on agricultural plant biomass and production. Rice is a valuable crop that is susceptible to salt and high temperatures. Here, we studied the role of flavanol 3-hydroxylase in response to combined salt and heat stress with the aim of better understanding the defensive mechanism of rice. We found that, compared with wild-type plants, the growth and development of transgenic plants were improved due to higher biosynthesis of kaempferol and quercetin. Furthermore, we observed that oxidative stress was decreased in transgenic plants compared with that in wild-type plants due to the reactive oxygen species scavenging activity of kaempferol and quercetin as well as the modulation of glutathione peroxidase and lipid peroxidase activity. The expression of high-affinity potassium transporter (HKT) and salt overly sensitive (SOS) genes was significantly increased in transgenic plants compared with in control plants after 12 and 24 h, whereas sodium-hydrogen exchanger (NHX) gene expression was significantly reduced in transgenic plants compared with in control plants. The expression of heat stress transcription factors (HSFs) and heat shock proteins (HSPs) in the transgenic line increased significantly after 6 and 12 h, although our understanding of the mechanisms by which the F3H gene regulates HKT, SOS, NHX, HSF, and HSP genes is limited. In addition, transgenic plants showed higher levels of abscisic acid (ABA) and lower levels of salicylic acid (SA) than were found in control plants. However, antagonistic cross talk was identified between these hormones when the duration of stress increased; SA accumulation increased, whereas ABA levels decreased. Although transgenic lines showed significantly increased Na+ ion accumulation, K+ ion accumulation was similar in transgenic and control plants, suggesting that increased flavonoid accumulation is crucial for balancing Na+/K+ ions. Overall, this study suggests that flavonoid accumulation increases the tolerance of rice plants to combined salt and heat stress by regulating physiological, biochemical, and molecular mechanisms.

Stevioside Attenuates Insulin Resistance in Skeletal Muscle by Facilitating IR/IRS-1/Akt/GLUT 4 Signaling Pathways: An In Vivo and In Silico Approach.

Type-2 diabetes mellitus (T2DM), the leading global health burden of this century majorly develops due to obesity and hyperglycemia-induced oxidative stress in skeletal muscles. Hence, developing novel drugs that ameliorate these pathological events is an immediate priority. The study was designed to analyze the possible role of Stevioside, a characteristic sugar from leaves of Stevia rebaudiana (Bertoni) on insulin signaling molecules in gastrocnemius muscle of obesity and hyperglycemia-induced T2DM rats. Adult male Wistar rats rendered diabetic by administration of high fat diet (HFD) and sucrose for 60 days were orally administered with SIT (20 mg/kg/day) for 45 days. Various parameters were estimated including fasting blood glucose (FBG), serum lipid profile, oxidative stress markers, antioxidant enzymes and expression of insulin signaling molecules in diabetic gastrocnemius muscle. Stevioside treatment improved glucose and insulin tolerances in diabetic rats and restored their elevated levels of FBG, serum insulin and lipid profile to normalcy. In diabetic gastrocnemius muscles, Setvioside normalized the altered levels of lipid peroxidase (LPO), hydrogen peroxide (H2O2) and hydroxyl radical (OH*), antioxidant enzymes (CAT, SOD, GPx and GSH) and molecules of insulin signaling including insulin receptor (IR), insulin receptor substrate-1 (IRS-1) and Akt mRNA levels. Furthermore, Stevioside enhanced glucose uptake (GU) and oxidation in diabetic muscles by augmenting glucose transporter 4 (GLUT 4) synthesis very effectively in a similar way to metformin. Results of molecular docking analysis evidenced the higher binding affinity with IRS-1 and GLUT 4. Stevioside effectively inhibits oxidative stress and promotes glucose uptake in diabetic gastrocnemius muscles by activating IR/IRS-1/Akt/GLUT 4 pathway. The results of the in silico investigation matched those of the in vivo study. Hence, Stevioside could be considered as a promising phytomedicine to treat T2DM.

Outcome of COVID-19: Diabetes and Obesity

Today the world is facing one of the biggest crisis, due to a new beta-corona virus emerged from Wuhan in China, on December 2019. WHO declared ‘acute respiratory syndrome COVID-19 (SARS Co-2)’as a pandemic on March 12, 2020. Corona viruses are enveloped positive single stranded RNA viruses, causing severe acute respiratory syndrome in the infected individuals. The risk of getting infected by covid-19 is similar in all the individuals across the nation. But the outcome of the infection varies from one individual to another, depending on the comorbidities present in them. The most vulnerable group of patients in respect to severity of outcome of the infection are the once with unbalanced heath conditions like age (&gt;65 years), immune-compromised, hypertension, type 2 diabetes, increased insulin resistance, cardiovascular diseases, chronic kidney disease, chronic liver disease, vasculitis(vascular inflammation)and obesity. It is now a public knowledge that diabetes and obesity are a risk factor for any individual as these conditions can exacerbate the manifestations of COVID-19 infections thus increasing the severity of the condition, that may require hospitalization of the patient, later may even require intensive care unit or/and mechanical ventilation, with increased risk of mortality rates. In diabetic patients it is mainly due to failure in controlling the glucose levels and the risk of ketoacidosis. In patients with obesity lipid peroxidase creates reactive lipid aldehydes leading to poor prognosis.

Moringa oleiferamitigates oxytetracycline stress inOreochromis niloticus

Antibiotics serve as the cornerstone of the treatment of bacterial diseases in the aquaculture sector. In this study, 210 Nile tilapias (Oreochromis niloticus) weighing approximately 60 ± 2.5 g b.w. were randomly divided into seven treatments (T1–7). The fish were experimentally infected with Aeromonas hydrophila and then treated with oxytetracycline (OTC) after the appearance of clinical symptoms. The fish were fed Moringa oleifera diet alone or M. oleifera diet before OTC treatment to alleviate the effects of OTC treatment. The effects of OTC treatment and M. oleifera were assessed by measuring the gene expressions of cytokines (interleukin [IL]-1β, IL-6, tumour necrosis factor-α, IL-10 and heat shock protein-70), antioxidant enzymes (catalase, superoxide dismutase, lipid peroxidase and total antioxidant capacity) and liver enzymes (aspartate aminotransaminase, alanine aminotransaminase and alkaline phosphatase). Although OTC treatment protected fish from A. hydrophila, leading to a decreased mortality rate (49.96%), it exerted deleterious effects on their immune and oxidative statuses. Thus, we recommend that M. oleifera be included in the feed of O. niloticus, especially those exposed to bacterial infection.

Obtaining Salt Stress-Tolerant Eggplant Somaclonal Variants from In Vitro Selection.

An efficient regeneration protocol was applied to regenerate shoots on salt stress-tolerant calli lines of aubergine (Solanum melongena). These NaCl-tolerant cell lines were obtained by two different methods. On the one hand, the developed callus tissue was transferred to a medium with a continuous salt content of 40, 80, 120, or 160 mM NaCl. On the other hand, the callus tissue was subjected to a stepwise increasing salinity to 160 mM NaCl every 30 days. With the second method, calli which could be selected were characterized by compact growth, a greenish color, and absence of necrotic zones. When grown on salt-free medium again, NaCl-tolerant calli showed a decline in relative growth rate and water content in comparison to the control line. This was more obvious in the 120 mM NaCl-tolerant callus. Lipid peroxidase activity increased in 40 and 80 mM NaCl-tolerant calli; yet did not increase further in 120 mM-tolerant callus. An increase in ascorbic acid content was observed in 80 and 120 mM NaCl-tolerant calli compared to the 40 mM NaCl-tolerant lines, in which ascorbic acid content was twice that of the control. All NaCl-tolerant lines showed significantly higher superoxide dismutase (SOD) (208–305–370 µmol min−1 mg−1 FW) and catalase (CAT) (136–211–238 µmol min−1 mg−1 FW) activities compared to control plants (231 and 126 µmol min−1 mg−1 FW). Plants were regenerated on the calli lines that could tolerate up to 120 mM NaCl. From the 32 plants tested in vitro, ten plants with a higher number of leaves and root length could be selected for further evaluation in the field. Their high salt tolerance was evident by their more elevated fresh and dry weight, their more increased relative water content, and a higher number and weight of fruits compared to the wild-type parental control. The presented work shows that somaclonal variation can be efficiently used to develop salt-tolerant mutants.

Evaluation of insecticidal potential of organochemicals on SF9 cell line

BackgroundOrganophosphates and Pyrethroids are the most widely used pesticides worldwide and are known to have significant toxicity on the nervous system of the target pest. Assessment for combined toxicity of Organophosphate and Pyrethroid on Sf9 (Spodoptera frugiperda) cells is less explored. The present study demonstrates and compares the two organochemicals whose trade names are Ammo and Profex, for its cytotoxic potential on the insect Sf9 cells. Ammo and Profex were selected as the test chemicals as toxicity of these insecticides at molecular and cellular level is poorly understood.ResultsThe results of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay demonstrated that Ammo and Profex exhibited significant cytotoxicity to Sf9 cells in a time- and dose-dependent manner. In our study, the IC50 value was obtained by MTT assay and the sub-lethal concentrations (IC50/20-17.5 µg/ml, IC50/10-35 µg/ml, and IC50/5–70 µg/ml for Ammo and IC50/20-20 µg/ml, IC50/10-40 µg/ml, and IC50/5-80 µg/ml for Profex) were selected for further tests. Acridine orange/ethidium bromide staining proved the apoptotic cell death on exposure of both the insecticides confirming its toxic potential. Furthermore, antioxidant status was assessed using DCF-DA staining and both the insecticides resulted into an increased reactive oxygen species (ROS) generation. A dose- and time-dependent significant (p < 0.05) alterations in lipid peroxidase (LPO), glutathione (GSH) and catalase (CAT) activity were observed.ConclusionThe results showed that both Ammo and Profex triggered apoptosis in Sf9 cells through an intrinsic mitochondrial pathway via the generation of ROS. Of the two insecticides, Ammo was found to be more toxic compared to Profex. The present study is important to evaluate the environmental safety and risk factors of Organochemicals’ exposure to crops and livestock.

Unveiling antioxidant and anti-cancer potentials of characterized Annona reticulata leaf extract in 1,2-dimethylhydrazine-induced colorectal cancer in Wistar rats

BackgroundColorectal cancer (CC) is the third most common cancer in the world. Annona reticulata (AR) also known as bullock's heart, is a traditional herb. AR leaf extract was initially investigated for its anti-bacterial, anti-inflammatory, anti-malarial, anti-helminthic, anti-stress, and wound healing properties. Only a few in vitro cancer studies have been conducted on AR. Although few studies have linked AR leaf extract to many cancers, comprehensive studies addressing regulation, biological functions, and molecular mechanisms leading to CC pathogenesis are clearly lacking. ObjectivesThe present study aimed to explore the antioxidant and anti-cancer potentials of AR leaf extract in CC. Materials and methodsThe MTT assay was used to test the anti-proliferative activity of AR leaf extract in vitro on the HCT116 cell line. Qualitative and quantitative phytochemical characterization was carried out using gas chromatography: mass spectrometry (GC–MS). 1,2-dimethylhydrazine (DMH) was used to establish CC model in female Wistar rats. The acute toxicity of AR leaf extract was tested in accordance with OECD guidelines. Aberrant Crypt Foci (ACF) count, organ index, and hematological estimations were used to screen for in vivo anti-cancer potential. The antioxidant activity of colon homogenate was determined. ResultsThe alcoholic leaf extract (IC50, 0.55 μg/ml) was found to be more potent than the aqueous extract. Using GC–MS, a total of 108 compounds were quantified in the alcoholic leaf extract. The LD 50 value was found to be safe at a dose of 98.11 mg/kg of body weight. AR alcoholic leaf extract significantly (p < 0.05) decreased ACF count and normalized colon length/weight ratio. AR leaf extract increased RBC, hemoglobin and platelets levels. The AR alcoholic leaf extract reduced the DMH-induced tumors and significantly (p < 0.05) increased the activity of endogenous antioxidant enzymes such as catalase, reduced glutathione, superoxide dismutase, and decreased the lipid peroxidase activity. AR leaf extract reduced the inflammation caused by DMH and helped to repair the colon's damaged muscle layers. ConclusionBased on the findings from the present study, it can be concluded that the alcoholic leaf extract of AR has antioxidant and anti-proliferative properties and can aid in the prevention of CC development and dysplasia caused by DMH.

Lipid Peroxidation Level and Peroxidase Activity in (Vitis vinifera L.) Leaves Infected with Grapevine Leafroll-Associated Virus 3

Objective: Grapevine (Vitis vinifera L.) is one of the major crops widely cultivated for the wine industry, as well as for the production of fresh and dried fruit in Azerbaijan. Grapevine leafroll disease (GLD) affects the vines throughout the world and is considered the most economically destructive among all virus and virus-like diseases. Vineyard surveys were conducted to determine the virus infection in the major grapevine growing region of Azerbaijan during 2019-2020. Methods/Statistical analysis: Forty-six samples were collected from grapevine fields and screened by rapid one-step assay AgriStrip and double-antibody sandwich Enzyme-linked immunosorbent assay (DASELISA). In our study, the levels of malondialdehyde (MDA), hydrogen peroxide (H2O2), pigments, relative water content (RWC), alterations in the activities of peroxidase enzymes (the activities of ascorbate peroxidase (APO), benzidine peroxidase (BPO) and guaiacol peroxidase (GPO) were investigated. Findings: The results revealed that tested samples were infected with Grapevine leafrollassociated virus 3 (GLRaV-3), however, no sample was found infected with other Grapevine leafroll-associated viruses. Changes in the levels of MDA, H2O2, pigments, RWC, alterations in the activities of peroxidase enzymes were studied. Significant reduction in green pigments like chlorophylls (a, b and total) and a gradual reduction in carotenoids were observed in all infected species. Obtained results showed that the level of RWC and the amount of hydrogen peroxide had increased in all infected leaves. The activities of APO, BPO and GPO were observed to increase in virus-infected leaves compared to the healthy control. Based on the results, it can be concluded that the effect of GLRaV-3 was more destructive in the N2 sample. The antioxidant defense system works more effectively in the N5 sample and this plant is more resistant to viral infection. Novelty: These results indicate that GLRaV-3 was the only most prevalent endemic viral pathogen of grapevine in the dominantly warmhumid continental climate of our country. All variable physiological parameters are assessed as the plant’s response to the pathogen. Keywords: activity; chlorophyll; hydrogen peroxide; malondialdehyde; peroxidase; virus disease

CRISPR/Cas9 mediated gene correction ameliorates abnormal phenotypes in spinocerebellar ataxia type 3 patient-derived induced pluripotent stem cells

Spinocerebellar ataxia type 3/Machado–Joseph disease (SCA3/MJD) is a progressive autosomal dominant neurodegenerative disease caused by abnormal CAG repeats in the exon 10 of ATXN3. The accumulation of the mutant ataxin-3 proteins carrying expanded polyglutamine (polyQ) leads to selective degeneration of neurons. Since the pathogenesis of SCA3 has not been fully elucidated, and no effective therapies have been identified, it is crucial to investigate the pathogenesis and seek new therapeutic strategies of SCA3. Induced pluripotent stem cells (iPSCs) can be used as the ideal cell model for the molecular pathogenesis of polyQ diseases. Abnormal CAG expansions mediated by CRISPR/Cas9 genome engineering technologies have shown promising potential for the treatment of polyQ diseases, including SCA3. In this study, SCA3-iPSCs can be corrected by the replacement of the abnormal CAG expansions (74 CAG) with normal repeats (17 CAG) using CRISPR/Cas9-mediated homologous recombination (HR) strategy. Besides, corrected SCA3-iPSCs retained pluripotent and normal karyotype, which can be differentiated into a neural stem cell (NSCs) and neuronal cells, and maintained electrophysiological characteristics. The expression of differentiation markers and electrophysiological characteristics were similar among the neuronal differentiation from normal control iPSCs (Ctrl-iPSCs), SCA3-iPSCs, and isogenic control SCA3-iPSCs. Furthermore, this study proved that the phenotypic abnormalities in SCA3 neurons, including aggregated IC2-polyQ protein, decreased mitochondrial membrane potential (MMP) and glutathione expressions, increased reactive oxygen species (ROS), intracellular Ca2+ concentrations, and lipid peroxidase malondialdehyde (MDA) levels, all were rescued in the corrected SCA3-NCs. For the first time, this study demonstrated the feasibility of CRISPR/Cas9-mediated HR strategy to precisely repair SCA3-iPSCs, and reverse the corresponding abnormal disease phenotypes. In addition, the importance of genetic control using CRISPR/Cas9-mediated iPSCs for disease modeling. Our work may contribute to providing a potential ideal model for molecular mechanism research and autologous stem cell therapy of SCA3 or other polyQ diseases, and offer a good gene therapy strategy for future treatment.

Evaluation of the effects of 5-fluorouracil and cyclophosphamide on Lathyrus sativus L.

BackgroundThe effect of 5-fluorouracil (5FU) and cyclophosphamide (CP) on grass pea (Lathyrus sativus L.) has been studied.ResultsThe effect of the selected drugs on radicle length, colchicine induces polyploidy and in vitro callus growth has been studied (concentration used: 5-Fluorouracil—0.01, 0.1, 1 and 10 mM; Cyclophosphamide—1, 10, 20 and 30 mM). The biochemical studies on superoxide dismutase, catalase and lipid peroxidase activity also studied. Radicle length, polyploidy percentage and callus growth decrease in both the drugs in a dose-dependent manner. The SOD, catalase and LP activity decrease with the increase in drug concentration except for low dose (for 5FU—0.01 mM and 1 mM for CP). Induced polyploidy (Control B) than water germinated seedling (Control A) shows higher enzyme activity but a decrease in the increased dose of drugs.ConclusionsThe present work has been done to assess the effective potentiality of two anticancerous drugs 5FU and CP with an objective to establish plant system as a model for preliminary screening of anticancerous lead compounds. The result of the present work would pave the way for the screening of unknown lead compounds with the potentiality to act as base analogue and DNA cross-linking drugs. This system is faster, cost-effective and convenient than animal model.

Antioxidant enzymes and non-enzymatic antioxidants as defense mechanism of salinity stress in cowpea (Vigna unguiculata L. Walp)\u2014Ife brown and Ife bpc

BackgroundSeveral mechanisms had been exhibited by plants to mitigate deleterious effects of salinity stress. A screen house experiment was conducted to investigate the effects of salinity stress on the activities of osmolytes (antioxidative and non-antioxidative enzymes) in the leaves of two cowpea (Vigna unguiculata L. Walp)—Ife brown and Ife bpc, with the aim of better understanding the biochemical mechanisms of salt tolerance. Salts of sodium chloride (NaCl) and sodium sulfate (Na2SO4) at 5, 10 and 15 dS/m concentrations were used for this study. The saline solution was prepared following standard methods. Proline, lipid peroxidase (LP), superoxide dismutase (SOD) and glutathione (GSH) were determined following standard protocols.ResultsResults showed that minimum proline content (12.07 mg/g) and maximum proline determination (16.05 mg/g) were observed in Ife bpc at 5 and at 15 dS/m under NaCl and Na2SO4 treatments. The LP content significantly increased in Ife brown at 15 dS/m under NaCl treatment and at 10 dS/m (9.49 mg/g) under Na2SO4 salinity. Minimum GSH content (120 µm/g) and maximum glutathione accumulation (138.97 µm/g) were observed in Ife bpc in the stressed cowpea seedlings (5 and 10 dS/m) under NaCl treatment with respect to the control. Also, SOD activities in the leaves of Ife brown increase with increase in salinity stress in both NaCl and Na2SO4 treatments.ConclusionsThis study concludes that the accumulation of enzymatic and non-enzymatic antioxidants is capable of detoxifying and scavenging reactive oxygen species, thereby mitigating salinity-induced oxidative damage.

Effect of gallic acid and Terminalia chebulaon hepatic oxidative stress markers of high fat diet induced hyperlipidemicmice

Introduction and Aim:Hyperlipidemia is caused due to lipid abnormalities leading to lipid peroxidation which causes extensive damage to hepatic tissues. The aim of the study is to induce hyperlipidemia in mice using high fat diet (HFD) and to estimate the effect of gallic acid and Terminalia chebula in reducing hepatic oxidative stress caused by hyperlipidemia. Materials and Methods:36 C57BL/6J Male18 ±3 g was divided into four groups; group I(6): received normal diet for 16 weeks, group II(10). High fat diet (HFD) + 0.5% CMC , group III (10)received HFD + gallic acid (50mg/Kg/b.wt) and group IV(10) HFD + T. chebula(250mg/kg/ b.wt)only normal diet for 15 weeks. Tissue samples from liver were taken for determination of reduced glutathione (GSH) levels catalase (CAT), super oxide dismutase (SOD) and Lipid peroxidase(LPO). Results:The data revealed that feeding the HFD diet significantly raised final body weight and caused hyperlipidemia, which elevated oxidative stress markers.The treatment given to mice HFD + T. chebula (250mg/kg) group IV showed significant increase in antioxidant enzymesand reduced acid reactive substances in blood GSH (47 %p<0.001), CAT (80 % p<0.001),SOD (55 % p<0.001)&LPO (48 % p<0.001) than mice of group III HFD + gallic acid (50mg/Kg/b.wt) GSH (32 % p<0.001), CAT (40 % p<0.001),SOD (31 % p<0.001)&LPO (38 % p<0.01). Conclusion:High fat diet-induced hyperlipidemia is coupled with increased hepatic oxidative stress, which is characterized by a decrease in antioxidant enzymes activityand increased lipid peroxidationis being more effectively treated by T. chebula powder than gallic acid (pure compound).