Current Status and Future Perspectives on Distribution of Fungal Endophytes and Their Utilization for Plant Growth Promotion and Management of Grapevine Diseases.

Grapevine is one of the economically most important fruit crops cultivated worldwide. Grape production is significantly affected by biotic constraints leading to heavy crop losses. Changing climatic conditions leading to widespread occurrence of different foliar diseases in grapevine. Chemical products are used for managing these diseases through preventive and curative application in the vineyard. High disease pressure and indiscriminate use of chemicals leading to residue in the final harvest and resistance development in phytopathogens. To mitigate these challenges, the adoption of potential biocontrol control agents is necessary. Moreover, multifaceted benefits of endophytes made them eco-friendly, and environmentally safe approach. The genetic composition, physiological conditions, and ecology of their host plant have an impact on their dispersion patterns and population diversity. Worldwide, a total of more than 164 fungal endophytes (FEs) have been characterized originating from different tissues, varieties, crop growth stages, and geographical regions of grapevine. These diverse FEs have been used extensively for management of different phytopathogens globally. The FEs produce secondary metabolites, lytic enzymes, and organic compounds which are known to possess antimicrobial and antifungal properties. The aim of this review was to understand diversity, distribution, host-pathogen-endophyte interaction, role of endophytes in disease management and for enhanced, and quality production.

Automated, cryogen-free headspace-trap with gas chromatography-mass spectrometry analysis of ethylene oxide and 2-chloroethanol as residual fumigants in foods.

Ethylene oxide (EtO), although banned for use, is still being detected in foodstuffs that have been fumigated to eradicate pests during storage and transport. Residual levels over the European Union's (EU) maximum residue limit (MRL) pose severe health concerns. Recent detection of EtO and its by-product 2-chloroethanol (2-CE) at alarming levels have led to product recalls throughout the EU. Here, a simple, automated headspace (HS)-trap method for the simultaneous determination of EtO and its derivative 2-CE by gas chromatography-mass spectrometry (GC-MS) at the required MRL of ≤ 0.05 mg/kg has been implemented. Syringe-based HS combined with backflushed trapping technology provided enrichment of multiple extractions from the same sample vial (known as multi-step enrichment or MSE®) to increase sensitivity for EtO and 2-CE analysis by GC-MS using single-ion-monitoring (SIM) mode. Method detection limits (MDLs) of 0.00059 mg/kg and 0.00219 mg/kg for EtO and 2-CE, respectively, were obtained without the need for manual handling, solvent extraction or derivatization methods. Recoveries were shown to average (n = 5) at 98% and 107% for EtO and 2-CE, respectively, and the reproducibility was <10% for both compounds.

Multiresidue analysis of pesticides in three Indian soils: method development and validation using gas chromatography tandem mass spectrometry.

The paper reports a multiresidue method that was validated on 220 multi-class pesticides in three major Indian soils, namely, (i) new alluvial soil (NAS); (ii) red lateritic soil (RS) and (iii) black soil (BS) from three different regions. An ethyl acetate-based extraction method with a freezing-out cleanup step was employed for sample preparation, followed by gas chromatography-tandem mass spectrometric analysis. The method that was initially optimized on BS worked satisfactorily for the other two soil matrices. At the spiking level of 10 µg/kg (LOQ), the recoveries were satisfactory (within 70-120%) with precision-RSDs, ≤20% (n = 6) for 85, 88.6, and 89% of compounds in BS, RS, and NAS respectively. At 20 µg/kg, the method performance was satisfactory in each soil for all pesticides. When this validated method was applied to analyse 25 field samples, 6 pesticides were detected in them. In each case, precision (RSD) was <20%. The method sensitivity, accuracy and precision complied with the SANTE/2020/12830 guidelines. The method can be applied for environmental monitoring and risk assessment purposes, thus aiding in regulating pesticide usage in agricultural fields. The limitations and future scope of the study are also discussed.HighlightsA multiresidue method is reported for simultaneous analysis of multi-class pesticides in diverse soilsThe method was validated on 220 pesticides in new alluvial, red lateritic and black soilsSample preparation involved extraction with ethyl acetate and cleanup by a freezing stepThe residues were estimated by gas chromatography tandem mass spectrometry (GC-MS/MS)The method accuracy and precision complied with the EU's SANTE guidelines.

Method development and validation in the curry leaf matrix employing advanced mass spectrometry: quantitative screening of 490 multiclass pesticides by buffered ethyl acetate technique.

Curry leaf is an evergreen herb with culinary, pharmaceutical, and nutraceutical applications. As pesticide residue in curry leaf has garnered significant regulatory attention in recent years, here we report a reliable method, which was validated for the determination of 265 and 225 pesticides using LC-MS/MS and GC-MS/MS, respectively. At first, the sample was comminuted after adding water (1:2). The sample preparation workflow included extraction of 10 g homogenized sample with 10 mL ethyl acetate (+1% acetic acid), cleanup by dispersive solid phase extraction (d-SPE, 50 mg PSA + 50 mg C18 + 10 mg GCB + 150 mg Na2SO4) and the final analysis by tandem mass spectrometry. The cleanup step adeptly removed co-extractives. The method effectively reduced matrix effects and offered an LOQ of 0.01 mg kg-1 for most compounds. The method's accuracy and precision results fulfilled the requirements of SANTE/11312/2021 guidelines at 0.01 mg kg-1 and higher levels of fortification. The accuracy and precision results were comparable for all pesticides. The successful screening of market samples indicates its high extraction efficiency and precision for incurred residue analysis. Due to its robustness and conformity with regulatory criteria, food testing laboratories worldwide can use the method to monitor pesticide levels in curry leaves.

Dynamic headspace GC-MS/MS analysis of ethylene oxide and 2-chloroethanol in dry food commodities: a novel approach.

With frequent RASFF notifications from the EU countries, the residue testing of ethylene oxide (EtO) and its metabolite 2-chloroethanol (2-CE) in food commodities has become essential to check their compliance with MRLs. This study, for the first time, aimed at establishing a dynamic headspace-GC-MS/MS method for the simultaneous determination of these two analytes in acetonitrile extracts of cumin, ashwagandha, chilli powder, turmeric powder, guar gum, locust bean gum, and ginger powder. The samples (4 g) were extracted using acetonitrile (10 mL). A dispersive-solid phase extraction cleanup step with primary secondary amine sorbent (50 mg/mL) reduced the interfering signal of (matrix-derived) acetaldehyde by >40% in chilli powder, ginger, turmeric, and guar gum. This cleanup was not required for sesame seeds. With high selectivity and sensitivity, the GC-MS/MS approach identified and quantified both compounds simultaneously. At the spiking levels of 0.01, 0.02, and 0.05 mg/kg, the recoveries and precision were satisfactory (70-120%, RSDs, ≤15%). The headspace method-performance was similar to liquid injections. The method provided reproducible results when evaluated by two different laboratories. The method provided high-precision results for incurred residue analysis. Given its efficiency, the validated method is anticipated to improve the effectiveness of monitoring of EtO residues in food commodities.

Regulatory landscape of risk assessment of pesticide residues in processed foods in India: a perspective.

In India, the levels of pesticide residues in Raw Agricultural Commodities (RAC) are being subjected to adequate legal regulations, and the health-risks associated with them are determined from time to time adhering to global standards. Since RACs are generally consumed by humans as the processed foods (PF), it is imperative to monitor the levels of pesticide residues in them in order to approach a realistic analysis of dietary exposure and concomitant health risk assessment. In India, production and consumption of PFs have a rising trend and hence it is indispensable to monitor the residue levels of pesticides in largely consumed PFs. Depending on the processing methods and physicochemical properties of pesticides, the residue levels may decrease or increase in a PF when compared to the corresponding RAC. While obtaining data on processing factors (Pf), it is pragmatic to focus on those situations in which the residues get concentrated following the processing step. Currently, regulatory agencies of several countries and the CODEX have determined the levels of pesticide residues in processed agriculture commodities, arrived at the Pfs, and fixed the maximum residue levels. Since consumption of PFs in India has tremendously increased in recent times and there is paucity of data about their health risks/benefits, it is imminent to deliberate on the complexities associated with the issues of adopting the Pfs generated by other regulatory agencies and subsequently examine the possibilities of generating the required data on Pfs on a priority basis to enable a comprehensive risk assessment.

Multiresidue analysis of pesticides, polyaromatic hydrocarbons and polychlorinated biphenyls in poultry meat and chicken eggs by GC-MS/MS: method development and validation.

The study uses gas chromatography with tandem mass spectrometry (GC-MS/MS) to develop a reliable analytical approach for detecting multiclass pesticides, polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) in poultry meat and chicken eggs. The meat (2 g) and egg (4 g) samples were extracted with acidified acetonitrile (10 mL) as part of the optimized sample preparation technique. The cleanup consisted of freezing an aliquot of the extract (5 mL) at -20 °C, followed by dispersive solid phase extraction using 50 mg PSA + 100 mg C18+150 mg MgSO4. The matrix co-extractives were effectively removed and the method performance met the European Commission's analytical quality control criteria (SANTE/12682/2019). The method was validated at two spiking levels (10 and 20 ng/g of 225 pesticides, 9 PAHs and 8 PCBs), and good recoveries (70-120%) and precision-RSDs (≤20%) were achieved for 90% of the targeted pesticide residues. For 80% of the compounds, the LOQs were ≤10 ng/g. The results of the intra-laboratory (involving six analysts) and inter-laboratory validation studies (involving eight ISO 17025 accredited laboratories) established satisfactory ruggedness and reproducibility. It created potential applications in commercial residue testing laboratories for regulatory compliance check purposes.

Bioefficacy, residue dynamics and dietary risk assessment of gibberellic acid in improving the potential yield of tomato (Solanum lycopersicum L.).

The increasing use of gibberellic acid (GA3) to promote fruit growth and yield has necessitated research into its trace level determination and estimation in harvested product. The phytohormone has increased the tomato yield (tonne ha-1) up to 24.7% with uniform fruit shape, size colour and lustre. A fast, simple, high-throughput analytical method was standardised based on electrospray ionisation - liquid chromatography-tandem mass spectrometry (LC-MS/MS). The samples were extracted using acidified (1% formic acid) methanol. The method was validated as per the SANTE/12682/2019 guidelines. The limits of detection (LOD) and quantification (LOQ) were 0.01 and 0.05 mg kg-1. The average recoveries at LOQ and higher levels were in the range of 86-108% with relative standard deviation (RSD) < 20%. The validated method was successfully applied under field condition by following first-order kinetics with half-lives (T1/2) 1.76 days (recommended dose) and 1.99 days (double dose). The estimated pre-harvest intervals (PHIs) were 6 days (recommended dose) and 8 days (double dose). Studies on dietary risk assessment concluded that even after spray of GA3 at recommended dose, the harvested produce (tomato) could be consumed safely.

Decontamination of aflatoxin B1 in peanuts using various cooking methods.

Peanut and its processed products are recurrently contaminated with aflatoxins (AFs) which are of potential public health concern. Among the different types of AFs, Aflatoxin B1 (B1) is the most frequently detected in peanuts over the maximum level (ML), and thus has warranted considerable research interest in the domain of food safety. In this study, we investigated the decontamination of B1 in three naturally-incurred lots (4, 12, and 40 µg/kg) of peanuts by a range of cooking treatments, including frying, pressure cooking, and roasting. B1 concentrations were determined by ultra-high performance liquid chromatography- fluorescence detection. The method provided a limit of quantification of 0.25 µg/kg for B1, which was much lower than any of its national and international MLs. The recoveries of B1 in fresh and cooked peanuts (positive-control) were in the range of 90-100%. Overall, all the cooking methods demonstrated a significant reduction in B1 loads. The degree to which the processing methods reduced the B1 content followed the pattern: roasting with a combination of NaCl and citric acid > pressure-cooking with a combination of NaCl and citric acid > frying. As the cooking procedures did not involve any complicated steps or sophisticated equipment, these could be readily adopted for decontamination or reduction in the level of B1 for a safer consumption of peanuts at the household level without affecting the organoleptic properties.

Coupling the high-resolution LC-MS characterisation of the phenolic compounds with the antimicrobial and antibiofilm properties of helencha (Enydra fluctuans Lour.).

This study reports the polyphenol profile of helencha (Enydra fluctuans Lour.), an underutilised, aquatic leafy vegetable, based on high resolution accurate mass analysis. The methanolic extract of helencha leaves was screened by ultra-high performance liquid chromatography with quadrupole time of flight mass spectrometry (LC-QToF-MS). An in-house developed database of phytochemical metabolites was referred for compound identifications. Based on the detection of the pseudomolecular ion and at least one molecule-specific fragment ion (each with < 5 ppm of mass error), 25 potentially-bioactive phenolic compounds were putatively identified. These included 6 flavonols, 4 phenolic acids, 3 lignans, 3 flavones and 1 each of flavanol, flavanone, dihydroflavonol, tetramethoxyflavone, isoflavonoid and methylated flavonol. In addition, 3 unclassified compounds are also reported. The helencha extract showed antibiofilm properties with a potent bacteriostatic activity against the clinical isolates of Pseudomonas aeruginosa, a human pathogenic bacteria. The complementary molecular docking studies indicated strong binding interactions of the identified compounds with the active site of LasR protein of P. aeruginosa. The in vitro and in silico study results would be useful to develop novel neutraceutical products based on helencha-extract and design new lead compounds to control the biofilm producing pathogenic microorganisms. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at (10.1007/s13197-021-04968-y).

Suitability of stationary phase for LC analysis of biomolecules.

Biologically active compounds such as carotenoids/isoprenoids, vitamins, steroids, saponins, sugars, long chain fatty acids, and amino acids play a very important role in coordinating functions in living organisms. Determination of those substances is indispensable in advanced biological sciences. Engineered stationary phase in LC for the analysis of biomolecules has become easier with the development of chromatographic science. In general, C18 column is being used for routine analysis but specific columns are being used for specific molecule. Monolithic columns are found to have higher efficiency than normal column. Among recent introduction, triacontyl stationary phases, designed for the separation of carotenoid isomers, are widely used for the estimation of carotenoids. In comparison to conventional C18 phases, C30 phases exhibited superior shape selectivity for the separation of isomers of carotenoids. It is also found useful for better elution and analysis of tocopherols, vitamin K, sterols, and fatty acids. Vitamin K, E, and their isomers are also successfully resoluted and analyzed by using C30 column. Amino bonded phase column is specifically used for better elution of sugars, whereas phenyl columns are suitable for the separation and analysis of curcuminoids and taxol. Like triacontyl stationary phase, pentafluorophenyl columns are also used for the separation and analysis of carotenoids. Similarly, HILIC column are best suited for sugar analysis. All the stationary phases are made possible to resolute and analyze the target biomolecules better, which are the future of liquid chromatography. The present article focuses on the differential interaction between stationary phase and target biomolecules. The applicability of these stationary phases are reported in different matrices.

Antityrosinase Activity of Combretum micranthum, Euphorbia hirta and Anacardium occidentale Plants: Ultrasound Assisted Extraction Optimization and Profiling of Associated Predominant Metabolites.

Tyrosinase is an important component of the enzyme polyphenol oxidase, which upon contact with the phenolic substrates forms the pigment melanin and induces undesirable food browning. The phenolic and triterpenoid compounds that naturally occur in plants are well known as tyrosinase inhibitors. Combretum micranthum (CM) leaves, Euphorbia hirta (EH) plant, and Anacardium occidentale (AO) fruits are traditionally known to have potential anti-tyrosinase activities. The aim of this study was to optimize the ultrasound-assisted extraction of secondary metabolites from these matrices, and to evaluate in tubo the antityrosinase activity of these extracts. Efforts were also taken to profile the secondary metabolites, mainly the phenolic and triterpenoid compounds, in order to understand their probable association with tyrosinase inhibition. The optimal ultrasound-assisted extraction conditions for simultaneous extraction of phenolic, and triterpenoid compounds were determined. The aqueous fraction of these extracts showed significant antityrosinase activity, with the CM leaves exhibiting the strongest inhibitory effect (IC50 of 0.58 g·L-1). The predominant metabolic compounds from these natural extracts were putatively identified by using a high-resolution quadrupole-time of flight (QToF) LC-MS instrument. The high-resolution accurate mass-based screening resulted in identification of 88 predominant metabolites, which included dihydrodaidzein-7-O-glucuronide, micromeric acid, syringic acid, morin, quercetin-3-O-(6″-malonyl-glucoside), 4-hydroxycoumarin, dihydrocaffeic acid-3-O-glucuronide, to name some, with less than 5 ppm of mass error.

Profiling of polyphenols in phalsa (Grewia asiatica L) fruits based on liquid chromatography high resolution mass spectrometry.

The aim of the study was to screen the metabolite profile of phalsa (Grewia asiatica), an underutilized fruit crop, using liquid chromatography-high resolution mass spectrometric analysis. A total of 50 compounds were tentatively identified based on their molecular mass and characteristic fragment ions, each with less than 5 ppm of mass error. These compounds included 21 flavonols, 2 dihydroflavonols, 7 flavones, 3 flavanols, 6 anthocyanins, 3 isoflavonoids, 2 phenolic acids, 2 flavanones, and 4 other phenolics. Flavonols were the predominant group of compounds, representing around 52.6% of the total phenolics. The paper has also discussed the potentiality of phalsa as an emerging functional food for the management of various human diseases in relation to the existing literature.

Volatome finger printing of red wines made from grapes grown under tropical conditions of India using thermal-desorption gas chromatography-mass spectrometry (TD-GC/MS).

The current study evaluated the key characters of aroma composition in diversified red wines (Cinsaut, Grenache, Cabernet Franc, Petit Verdot, Cabernet Sauvignon, Nielluccio, Tempranillo, Syrah, Merlot and Caladoc). Out of hundreds of volatile compounds 64 compounds were considered for study. Different groups consisting of fatty acids, volatile alcohols, aldehydes, esters, volatile phenols and terpenes were analysed using gas chromatography mass spectrometry coupled with thermal desorption (TD-GC-MS). Among all these diversified classes, alcohols were found as the most dominant group followed by esters and acids whereas aldehydes, phenols and terpenes were found to be minor compounds. Among the varieties, Nielluccio wine recorded highest concentration of total volatile compounds (191.53 mg/L) while, it was least in Caladoc wines (15.45 mg/L). The principal component analysis clearly differentiated Grenache wines based on their relationships between scores and their aroma composition followed by Nielluccio and Cinsuat wines. Out of sixty four compounds, only six aromatic compounds viz. butanediol, isoamyl actate, γ-Terpene, butanol, acetic acid and furfural have satisfying aroma descriptors with floral and fruity nuances and contribute to differentiate the Grenache wines from other varieties which have similar scores in PC1 analysis. The cluster analysis also suggested that the wines in the same group (Cinsaut, Tempranillo and Syrah), (Cabernet Franc, Cabernet Sauvignon, Caladoc and Merlot) and (Nielluccio and Petit Verdot) had similar aroma characterization. Grenache wines were well differentiated from the sub group formed by other red varieties.

Analytical method validation, dissipation and safety evaluation of combination fungicides fenamidone + mancozeb and iprovalicarb + propineb in/on tomato.

Dissipation behavior and degradation kinetics of fenamidone + mancozeb (Sectin 60 WG) and iprovalicarb + propineb (Melody Duo 66.75 WP) in tomato were studied at recommended dose (RD) and double dose (DD) of application. The analysis of the field samples were carried out by employing liquid chromatography tandem mass spectrometry (LC-MS/MS) for fenamidone and iprovalicarb residues and gas chromatography mass spectrometry for mancozeb and propineb residues after thorough validation of the extraction methods. The dissipation of residues best followed 1st + 1st order for all the test fungicides. The half-life period for fenamidone, mancozeb, iprovalicarb and propineb were 2, 2, 1.5 and 2 days for RD and 3, 2.5, 2 and 3 days for DD, respectively. The pre-harvest intervals were not applicable for iprovalicarb, fenamidone and mancozeb (at RD) as the residues at 0 day were below maximum residue limit set by European Union, and it was 1 day for DD of iprovalicarb, 3.5 days for DD of fenamidone, 3 days for DD of mancozeb, 3 and 7 days for propineb at RD and DD, respectively. A PHI of 4 and 7 days are proposed for fenamidone + Mancozeb and iprovalicarb + propineb, respectively. Dietary exposure calculated for all the pesticides were safe on all the sampling days except for propineb residues for which it was safe after first day of the double dose application. The study will be useful for promoting effective residue management and safe use of these chemicals for controlling fungal diseases in tomato crop.

Targeted phenolic profiling of Sauvignon blanc and Shiraz grapes grown in two regions of India by liquid chromatography-tandem mass spectrometry.

The phenolic compounds play an important role in production of quality grapes and wines. The current investigation focused on optimization of an extraction method for targeted analysis of 33 phenolic compounds in grapes by liquid chromatography tandem mass spectrometry (LC-MS/MS). The optimized method was successfully used for phenolic profiling of two wine grape varieties, Sauvignon blanc (white) and Shiraz (red) originated from Pune and Nasik regions of Maharashtra State, India. The optimized sample preparation procedure involved liquid-liquid extraction with acidified methanol by vortexing for 2 min followed by analysis on LC-MS/MS. The limit of quantification of the targeted compounds was in the range of 29 to 411 µg/L. The results indicated that skin of both varieties contained the highest amount of flavonols (69.47 ± 14.74 mg/kg in Sauvignon blanc and 129.47 ± 10.05 mg/kg in Shiraz) compared to pulp. The highest amounts of flavan-3-ols were present in grape seed collected from the Pune region (2016.84 ± 14.73 mg/kg in Sauvignon blanc and 1945.06 ± 32.69 mg/kg in Shiraz). The concentration of stilbenes was the highest in grape skin (0.13 ± 0.52 to 5.78 ± 5.45 mg/kg) compared to seed and pulp of both varities. Hydroxybenzoic acid (vanillin), hydroxycinnamic acid (p-coumaric acid) and anthocyanins (oenin, malvidin, cyanidin and kuromanin) were found only in Shiraz variety. The results of antioxidant activity (FRAP and DPPH assay) indicated the highest scavenging activity in seed (978.64 ± 56.23 to1133.38 ± 143.65 µMol TE/g DW FRAP and 594.93 ± 37.94 to 631.94 ± 56.45 µMol TE/g DW in DPPH). The phenolic contents in Sauvignon blanc and Shiraz grapes between Pune and Nasik regions did not have any significant difference.

High-sensitivity direct analysis of aflatoxins in peanuts and cereal matrices by ultra-performance liquid chromatography with fluorescence detection involving a large volume flow cell.

This paper reports a sensitive and cost effective method of analysis for aflatoxins B1, B2, G1 and G2. The sample preparation method was primarily optimised in peanuts, followed by its validation in a range of peanut-processed products and cereal (rice, corn, millets) matrices. Peanut slurry [12.5 g peanut + 12.5 mL water] was extracted with methanol: water (8:2, 100 mL), cleaned through an immunoaffinity column and thereafter measured directly by ultra-performance liquid chromatography-fluorescence (UPLC-FLD) detection, within a chromatographic runtime of 5 minutes. The use of a large volume flow cell in the FLD nullified the requirement of any post-column derivatisation and provided the lowest ever reported limits of quantification of 0.025 for B1 and G1 and 0.01 µg/kg for B2 and G2. The single laboratory validation of the method provided acceptable selectivity, linearity, recovery and precision for reliable quantifications in all the test matrices as well as demonstrated compliance with the EC 401/2006 guidelines for analytical quality control of aflatoxins in foodstuffs.

Chemo-profiling of anthocyanins and fatty acids present in pomegranate aril and seed grown in Indian condition and its bioaccessibility study.

The goal of the present study was to investigate the bioactive molecules (anthocyanins and fatty acids) present in the aril of pomegranate. Major anthocyanins present in the aril of pomegranate were identified by HRMS as delphinidin 3,5-diglucoside, cyanidin 3,5-diglucoside, pelargonidin 3,5-diglucoside, cyanidin 3-glucoside and delphinidin 3-glucoside. In-vitro study revealed that bioaccessibility of anthocyanin in duodenal condition was varied between 7.3 and 9.7%. Encapsulation enhances the bioaccessibility of both the phenolics to some extent in gastric as well as duodenal condition. Seed oil contains significant amount of unsaturated fatty acids especially ω-5 fatty acids. Geometrical isomers of ω-5 fatty acids were also identified by GC-MS. The spray dried anthocyanin formulation has potential for food application.

A newly synthesized nickel chelate can selectively target and overcome multidrug resistance in cancer through redox imbalance both in vivo and in vitro.

Induction of undesired toxicity and emergence of multidrug resistance (MDR) are the major obstacles for cancer treatment. Moreover, aggressive cancers are less sensitive towards existing chemotherapeutics. Therefore, selective targeting of cancers without inducing undesired side effects and designing proper strategies to overcome MDR has utmost importance in modern chemotherapy. Previously we revealed the anticancer properties of some transition metal chelates of Schiff base, but the effectiveness of nickel complex is still unrevealed. Herein, we synthesized and characterized a Schiff base nickel chelate, nickel-(II) N-(2-hydroxyacetophenone) glycinate (NiNG), through different spectroscopic means. NiNG proves to be a broad spectrum anticancer agent with considerable efficacy to overcome MDR in cancer. Antiproliferative effects of NiNG was evaluated using drug-resistant (CEM/ADR5000; NIH-MDR-G185; EAC/Dox), drug-sensitive aggressive (Hct116; CCRF-CEM; EAC/S) and normal (NIH-3T3) cells that reveal the selective nature of NiNG towards drug resistant and sensitive cancer cells without inducing any significant toxicity in normal cells. Moreover, NiNG involves reactive oxygen species (ROS)-mediated redox imbalance for induction of caspase 3-dependent apoptosis in aggressive drug-sensitive Hct116 and drug-resistant NIH-MDR-G185 cells through disruption of mitochondrial membrane potential. Moreover, intraperitoneal (i.p.) application of NiNG at non-toxic doses caused significant increase in the life-span of Swiss albino mice bearing sensitive and doxorubicin-resistant subline of Ehrlich ascites carcinoma cells. It is noteworthy that, in vitro NiNG can only overcome P-glycoprotein-mediated MDR while in vivo NiNG can overcome MRP1-mediated MDR in cancer. Therefore, NiNG has therapeutic potential to target and overcome MDR in cancer.

Modulation of cell death in human colorectal and breast cancer cells through a manganese chelate by involving GSH with intracellular p53 status.

Chemotherapy is central to current treatment modality especially for advanced and metastatic colorectal and breast cancers. Targeting the key molecular events of the neoplastic cells may open a possibility to treat cancer. Although some improvements in understanding of colorectal and breast cancer treatment have been recorded, the involvement of glutathione (GSH) and dependency of p53 status on the modulation of GSH-mediated treatment efficacy have been largely overlooked. Herein, we tried to decipher the underlying mechanism of the action of Mn-N-(2-hydroxyacetophenone) glycinate (MnNG) against differential p53 status bearing Hct116, MCF-7, and MDA-MB-468 cells on the backdrop of intracellular GSH level and reveal the role of p53 status in modulating GSH-dependant abrogation of MnNG-induced apoptosis in these cancer cells. Present study discloses that MnNG targets specifically wild-type-p53 expressing Hct116 and MCF-7 cells by significantly depleting both cytosolic, mitochondrial GSH, and modulating nuclear GSH through Glutathione reductase and Glutamate-cysteine ligase depletion that may in turn induce p53-mediated intrinsic apoptosis in them. Thus GSH addition abrogates p53-mediated apoptosis in wild-type-p53 expressing cells. GSH addition also overrides MnNG-induced modulation of phase II detoxifying parameters in them. However, GSH addition partially replenishes the down-regulated or modulated GSH pool in cytosol, mitochondria, and nucleus, and relatively abrogates MnNG-induced intrinsic apoptosis in p53-mutated MDA-MB-468 cells. On the contrary, although MnNG induces significant cell death in p53-null Hct116 cells, GSH addition fails to negate MnNG-induced cell death. Thus p53 status with intracellular GSH is critical for the modulation of MnNG-induced apoptosis.