Mahanimbine isolated from Murraya koenigii inhibits P-glycoprotein involved in lung cancer chemoresistance.

P-glycoprotein (P-gp), a transmembrane glycoprotein, is mainly involved in lung cancer multidrug resistance. Several P-gp inhibitors have been developed to enhance the efficacy of chemotherapeutics and overcome drug resistance. However, most of them failed in the clinical stages due to undesirable side effects. Therefore, there is a requirement to develop P-gp inhibitors from natural sources. Dietary spice bioactives have been well-known for their anticancer activities. However, their role in modulating the P-gp activity has not been well investigated. Therefore, we have screened for the potential bioactives from various spice plants with P-gp modulatory activity using computational molecular docking analysis. The computational analysis revealed several key bioactives from curry leaves, specifically mahanimbine, exhibited a strong binding affinity with P-gp. Unfortunately, mahanimbine is available with few commercial sources at very high prices. Therefore, we prepared a curry leaves extract and isolated mahanimbine by a novel, yet simple, extraction method that requires less time and causes minimum environmental hazards. After purification, structure, and mass were confirmed for the isolated compound by IR spectrum and LC-MS/MS analysis, respectively. In the mechanistic study, hydrolysis of ATP and substrate efflux by P-gp are coupled. Hence, ATP binding at the ATPase-binding site is one of the fundamental steps for the P-gp efflux cycle. We found that mahanimbine demonstrated to stimulate P-gp ATPase activity. Concurrently, it enhanced the intracellular accumulation of P-gp substrates Rhodamine 123 and Hoechst stain, which indicates that mahanimbine modulates the function of P-gp. In addition, we have analyzed the complementary effect of mahanimbine with the chemotherapeutic drug gefitinib. We found that mahanimbine synergistically enhanced gefitinib efficiency by increasing its intracellular accumulation in lung cancer cells. Overall, mahanimbine has been shown to be a potent P-gp modulator. Therefore, mahanimbine can be further developed as a potential candidate to overcome chemoresistance in lung cancer.

Cytotoxicity of Mahanimbine from Curry Leaves in Human Breast Cancer Cells (MCF-7) via Mitochondrial Apoptosis and Anti-Angiogenesis.

Mahanimbine (MN) is a carbazole alkaloid present in the leaves of Murraya koenigii, which is an integral part of medicinal and culinary practices in Asia. In the present study, the anticancer, apoptotic and anti-invasive potential of MN has been delineated in vitro. Apoptosis cells determination was carried out utilizing the acridine orange/propidium iodide double fluorescence test. During treatment, caspase-3/7,-8, and-9 enzymes and mitochondrial membrane potentials (Δψm) were evaluated. Anti-invasive properties were tested utilizing a wound-healing scratch test. Protein and gene expression studies were used to measure Bax, Bcl2, MMP-2, and -9 levels. The results show that MN could induce apoptosis in MCF-7 cells at 14 µM concentration IC50. MN-induced mitochondria-mediated apoptosis, with loss in Δψm, regulation of Bcl2/Bax, and accumulation of ROS (p ≤ 0.05). Caspase-3/7 and -9 enzyme activity were detected in MCF-7 cells after 24 and 48 h of treatment with MN. The anti-invasive property of MN was shown by inhibition of wound healing at the dose-dependent level and significantly suppressed mRNA and protein expression on MMP-2 and -9 in MCF-7 cells treated with a sub-cytotoxic dose of MN. The overall results indicate MN is a potential therapeutic compound against breast cancer as an apoptosis inducer and anti-invasive agent.

Imported edible leaves collected at retail sale in England during 2017 with an emphasis on betel and curry leaves: microbiological quality with respect to Salmonella, Shiga-toxin-producing E. coli (STEC) and levels of Escherichia coli.

AIMS: To investigate the microbiological quality of imported fresh leaves on retail sale during 2017 with respect to Salmonella, Shiga-toxin-producing Escherichia coli (STEC) and levels of E. coli. METHODS AND RESULTS: Two hundred and seventy-nine samples of imported edible leaves (69 banana, 77 betel, 118 curry and 15 other types) were tested. Salmonella spp. were confirmed by whole-genome sequencing and isolated from 44 samples, 26% from curry leaves, 14% from betel and 2·4% from all other leaf types: 80% of all samples contained ≥102 , 44% ≥103 and 22% ≥104 CFU of E. coli CFU per g. All samples where Salmonella were detected also yielded ≥20 CFU of E. coli/g. 54 samples were tested for STEC which was detected in six samples and isolated from three: one was identified as STEC O157:H7. CONCLUSIONS: This report further highlights an ongoing problem of Salmonella contamination of imported fresh edible leaves. SIGNIFICANCE AND IMPACT OF THE STUDY: Among all food tested by Public Health England (approximately 11 000 per annum), curry leaves were the herb most commonly contaminated with Salmonella, and betel leaves were the most commonly contaminated ready-to-eat food. The high proportion with unsatisfactory E. coli levels and the detection of STEC suggests risks of contamination by multiple enteric pathogens.

Bio-fortification and shelf-life extension of idli batter using curry leaves (Murraya koenigii).

Among several traditional foods of India, idli is one of the most popular and commonly consumed steamed products. A new method of adding Murraya koenigii (curry leaves) to idli batter as a vehicle for fortification and extension of shelf-life has been developed. Dried curry leaves powder was incorporated with other ingredients like rice and dehusked black gram in different proportions to optimize the most palatable formulation. Rate of fermentation and microbial changes in the batter; nutritional qualities, texture and sensory properties of the prepared product were assessed. Incorporation of curry leaves powder (5 %) in idli batter increased the shelf-life and also increased the flavour, texture and appearance of the idli. The calcium content of the prepared idli was 10 times more than that of the control idli, while dietary fiber content increased by 18.6 %. Anti-microbial activity of the curry leaves in idli batter extended the shelf-life from 2 to 5 days when stored at 30 °C.

An assessment of the microbiological safety of fresh whole-leaf herbs from retail premises in the United Kingdom with a focus on Salmonella spp.

AIMS: Fresh herbs have been associated with a number of outbreaks in recent years, in the United Kingdom and elsewhere. A study of fresh herbs was carried out to assess their microbiological safety in relation to Salmonella contamination and levels of Escherichia coli. METHODS AND RESULTS: Between January and March 2014, 774 samples of ready-to-eat, fresh, whole-leaf herbs were collected from retail premises in the United Kingdom. Overall, Salmonella was detected in nine samples (1·2%). Of these, five were curry leaves. Other herbs contaminated with Salmonella were basil (two samples), walleria (1) and coriander (1). Escherichia coli was detected in 13% of samples, with 11% containing unsatisfactory levels (≥10(2) g(-1)). CONCLUSIONS: Whilst 88% of samples in this study were of an acceptable microbiological quality, the presence of Salmonella and/or elevated E. coli levels in 12% is a cause for concern. Curry leaves, in particular, had significantly higher rates of contamination with both Salmonella and E. coli than other herbs. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights the potential public health risk associated with the consumption of certain ready-to-eat fresh herbs, and the need for good hygiene practices and effective decontamination procedures during the growth, harvesting and subsequent handling of these products.

Effect of curry leaves in lowering blood pressure among hypertensive Indian patients.

Hypertension, commonly known as high or rising blood pressure, is characterized by a consistently elevated blood arterial pressure. It is a significant medical condition called hypertension raises your risk of developing problems with your heart, brain, kidneys, and other organs. Curry leaves work well as a treatment for high blood pressure. Curry leaves are thought to have a potent therapeutic impact for the treatment of high blood pressure since they are low in salt and high in potassium. Therefore, it is of interest to reduce the level of blood pressure among adults in rural area with the help of curry leaves. According to the statistical analysis, the experimental group's post-test mean score for hypertension was 155 with a standard deviation of 13.892 while the control group's post-test mean score was 162 with a standard deviation of 17.20. The estimated unpaired value of t = 11.07 was deemed to be statistically significant at the p 0.05 level and the mean difference was 7. As a result, the null hypothesis was rejected and the research hypothesis was accepted. Giving Curry leaf powder to patients with hypertension as a consequence was an effective intervention.

Comparative antioxidant effect of aqueous extracts of curry leaves, fenugreek leaves and butylated hydroxytoluene in raw chicken patties.

Antioxidant properties and use of aqueous extracts of curry leaves (Murraya koenigii) and fenugreek leaves (Trigonella foenum-graecum) as source of natural antioxidant in raw chicken meat were evaluated. Four treatments viz., I. Control (meat +2% salt), II.BHT (meat +2% salt +0.1% BHT), III. CLE (meat +2% salt +2% curry leaves extract) and IV. FLE (meat +2% salt +2% fenugreek leaves extract) were compared for lipid oxidation during eight days refrigerated storage. The average phenolic content was 59.2 and 52.8 mg/g gallic acid equivalent in CLE and FLE respectively. Free radical scavenging activity was 61.4 and 64.2% in CLE and FLE, respectively. CLE had significantly (P < 0.05) higher reducing activity (2.4) as compared to FLE (2.2). Observation on lipid oxidation showed a significant (p < 0.05) higher TBARS values in control patties than other patties containing BHT, CLE and FLE. Overall percent increase during storage period was highest in control followed by BHT, CLE and FLE. Both synthetic antioxidants and natural extracts significantly (p < 0.05) decreased the TBARS values. The average percent decrease in TBARS values during storage was 18, 25.5 and 27.5 in BHT, CLE and FLE treatments, respectively. Therefore, it was concluded that water extracts obtained from curry leaves and fenugreek leaves could be explored as natural antioxidants in poultry meat and meat products.

Extract of Murraya koenigii selectively causes genomic instability by altering redox-status via targeting PI3K/AKT/Nrf2/caspase-3 signaling pathway in human non-small cell lung cancer.

BACKGROUND: Lung cancer is the leading cause of cancer-related death worldwide. Dietary bioactives have been used as alternative therapeutics to overcome various adverse effects caused by chemotherapeutics. Curry leaves are a widely used culinary spice and different parts of this plant have been used in traditional medicines. Curry leaves are a rich source of multiple bioactives, especially polyphenols and alkaloids. Therefore, extraction processes play a key role in obtaining the optimum yield of bioactives and their efficacy. PURPOSE: We aim to select an extraction process that achieves the optimum yield of bioactives in curry leaves crude extract (CLCE) with minimum solvent usage and in a shorter time. Further, to investigate the anticancer properties of CLCE and its mechanism against lung cancer. METHODS: Different extraction processes were performed and analyzed polyphenol content. The bioactives and essential oils present in curry leaves were identified through LC-MS/MS and GC-MS analysis. The cytotoxicity of microwave-assisted CLCE (MA-CLCE) was investigated through MTT and colony-forming assays. The DNA damage was observed by comet assay. The apoptotic mechanisms of MA-CLCE were investigated by estimating ROS production, depolarization of mitochondrial membrane potential (MMP), and apoptotic proteins. The glutathione assay estimated the antioxidant potential of MA-CLCE in normal cells. RESULTS: Generally, conventional extraction methods require high temperatures, extra energy input, and time. Recently, green extraction processes are getting wider attention as alternative extraction methods. This study compared different extraction processes and found that the microwave-assisted extraction (MAE) method yields the highest polyphenols from curry leaves among other extraction processes with minimum processing. The MA-CLCE functions as an antioxidant under normal physiological conditions but pro-oxidant to cancer cells. MA-CLCE scavenges free radicals and enhances the intracellular GSH level in alveolar macrophages in situ. We found that MA-CLCE selectively inhibits cell proliferation and induces apoptosis in cancer cells by altering cellular redox status. MA-CLCE induces chromatin condensation and genotoxicity through ROS-induced depolarization of MMP. The depolarization of MMP causes the release of cytochrome c into the cytosol and activates the apoptotic pathway in lung cancer cells. However, pretreatment with ascorbic acid, an antioxidant, inhibits the MA-CLCE-induced apoptosis by reducing ROS production, which impedes mitochondrial membrane disruption, preventing BAX/BCL-2 expression alteration. Simultaneously, MA-CLCE downregulates the expression of survival signaling regulator PI3K/AKT, which modulates Nrf-2. MA-CLCE also diminishes intracellular antioxidant proficiency by suppressing Nrf-2 expression, followed by HO-1 expressions. CONCLUSION: Among several extraction methods, MA-CLCE is rich in several bioactives, especially polyphenols, alkaloids, and essential oils. Here, we reported for the first time that MA-CLCE functions as a pro-oxidant to lung cancer cells and acts as an antioxidant to normal cells by regulating different cellular programs and signaling pathways. Therefore, it can be further developed as a promising phytomedicine against lung cancer.

The physicochemical, microbiological, and organoleptic properties and antioxidant activities of cream cheeses fortified with dried curry leaves (Murraya koenigii L.) powder.

We aimed to investigate the effects of dried curry leaves powder (CLP) incorporation on physicochemical, microbiological, antioxidant, and sensory properties of cream cheeses. Varying levels of CLP infusions (i.e., T1: 0% [control], T2: 0.15%, T3: 0.2%, and T4: 0.25%; w/w%) were stored for 10 days at 4°C. Antioxidant properties were evaluated using total phenolic content, 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging ability, and ferric reducing antioxidant power using in vitro assays. Total antioxidant capacity significantly (p < .05) increased with the increasing levels of CLP. Physicochemical and microbiological qualities were not significantly affected by the addition of CLP, indicating the suitability of using CLP without compromising the quality of cream cheese. Organoleptic properties were affected with CLP addition, where T3 had the highest scores for color, aroma, flavor, texture, and overall acceptability. The principal component analysis provides the holistic approach of studying the variation associated with cream cheeses and the overall relationship among studied parameters. This provides strong references for novel dairy products added with antioxidant-rich Murraya koenigii L. powder. The study also has merits to promote scientific knowledge concerning, and how the incorporation would influence the physicochemical, organoleptic, and microbiological properties of cream cheese to deliver the value-added or diversified product to emerging consumers.

Characterization of the Major Odor-Active Compounds in the Leaves of the Curry Tree Bergera koenigii L. by Aroma Extract Dilution Analysis.

Curry leaves are a popular seasoning herb with a pronounced sulfury and burnt odor, the molecular background of which was yet unclear. Application of an aroma extract dilution analysis to the volatile fraction of curry leaves isolated by solvent extraction and solvent-assisted flavor evaporation afforded 23 odor-active compounds with flavor dilution (FD) factors ranging from 1 to 8192. On the basis of the comparison of their retention indices, mass spectra, and odor properties with data of reference compounds, the structures of 22 odorants could be assigned, 15 of which had not been reported in curry leaves before. Odorants with high FD factors included 1-phenylethanethiol (FD factor 8192), linalool (4096), α-pinene (2048), 1,8-cineole (1024), (3Z)-hex-3-enal (256), 3-(methylsulfanyl)propanal (128), myrcene (64), (3Z)-hex-3-en-1-ol (32), and (2E,6Z)-nona-2,6-dienal (32). The unique sulfury and burnt odor exhibited by 1-phenylethanethiol in combination with its high FD factor suggested that it constitutes the character impact compound of fresh curry leaf aroma.

Protective effect of antioxidant rich aqueous curry leaf (Murraya koenigii) extract against gastro-toxic effects of piroxicam in male Wistar rats.

Piroxicam (chemically 4-hydroxy-2-methyl-N-2-pyridinyl-2H-1,2-benzothiazine-3-carboxamide), a classical non-steroidal anti-inflammatory drug (NSAID) is orally administered to arthritic patients. Inhibition of prostaglandin E2 (PGE2) synthesis and subsequent free hydroxyl radical generation in vivo exert gastro-toxic side effects on piroxicam treatment. Leaves of curry plant are rich in antioxidants with prolific free radical scavenging activities. This led us to investigate the efficiency of the use of curry leaves in ameliorating piroxicam induced gastric damage. Piroxicam was orally (30 mg per kg body weight) administered in male albino Wistar rats to generate gastric ulcers. These rats were orally fed with graded doses of aqueous extract of curry or Murraya koenigii leaves (Cu LE) prior to piroxicam administration. Oxidative stress biomarkers, activities of antioxidant and pro-oxidant enzymes, mucin content and nature, PGE2 level, activities of mitochondrial enzymes and histomorphology of gastric tissues were studied. Piroxicam treatment altered all the above mentioned parameters whereas, curry leaf extract pre-treated animals were protected against piroxicam induced alterations. Hence, the protective action of the antioxidant rich Cu LE was investigated to propose a new combination therapy or dietary management to arthritic patients using piroxicam.