Background: Triple Negative Breast Cancers (TNBCs) have high morbidity and shorter survival rate in the population. These types of cancers have high aggressiveness, lymphatic invasion, and absence of receptors. The treatment options for these types of cancers are also scarce. Several studies have been conducted to investigate the effectiveness of seeds of Annona muricata for its anti-cancer activities in various cancer cell lines, such as lung A549, breast MCF7, colon HT-29, oral KB, and human hepatoma cell lines. But works related to its anti-cancer effect and mechanism of action in TNBCs have not been elucidated. Objective: The present study was undertaken to evaluate the in vitro, in vivo, and in silico anti-cancer potential of chloroform fraction of methanolic extract of seeds of Annona muricata (CMAM) against TNBC along with elucidation of its mechanistic pathway. Methods: In vitro cytotoxicity- and antiproliferative- studies in three triple-negative breast cancer cell lines were conducted using the MTT and SRB assays, respectively. The mechanism through which CMAM exerts its pharmacological effect was elucidated in vitro employing cell morphological assessment studies using Acridine Orange/Ethidium Bromide (AO/EB), intracellular reactive oxygen species assay, DNA fragmentation assay, agarose gel electrophoresis, terminal deoxynucleotidyl Transferase dUTP Nick End Labeling (TUNEL) assay, cell cycle analysis, annexin binding assay, and caspase-activated mitochondria-mediated apoptotic assays using western blot. In vivo evaluation in 4T1 induced murine mammary tumor model was also conducted. Phytoconstituents in CMAM were analyzed using liquid chromatography mass spectroscopy. In silico binding studies with various annonaceous acetogenins against BCL-2 and cyclin E were performed. Results: Cytotoxicity studies in MDA-MD-231, 4TI, and BT-549 revealed the IC₅₀ value of CMAM to be 2.5±0.14, 4.8±0.3, and 4.5±0.16μg/mL, respectively. Anti-proliferative studies in 4T1, MDA-MB-231, and BT- 549 revealed the GI50 values to be 0.128+0.03, 18.03+0.20, 0.95+0.04μg/mL, respectively. CMAM exhibited its cytotoxicity through the lysis of cell membrane, ROS dependent caspase-activated mitochondria-mediated apoptosis, and arresting the S phase of the cell cycle. In vivo evaluation also supported the tumoricidal property of CMAM, as evidenced by a reduction in tumor volume and serum biomarkers. Histopathologically, there was a marked reduction in cellularity, nuclear chromatin condensation, and a few normal cells in the group treated with CMAM at a dose of 31mg/Kg. Phytoconstituent evaluation has revealed the presence of annonaceous acetogenins in CMAM. Among the various annonaceous acetogenins, muricatacin alone showed lipophilicity and binding affinity towards BCL-2 and cyclin E1. Conclusion: The current study shows the effectiveness of CMAM against TNBC both in vitro and in vivo. This anticancerous effect of CMAM could be by virtue of its ROS dependent caspase-activated mitochondriamediated apoptosis and the S-phase arrest of the cell cycle in the TNBCs. Our results indicate that the presence of annonaceous acetogenins, especially muricatacin, could be contributing to this anticancerous effect of CMAM. Thus, muricatacin could be a potential candidate for the targeted therapy of TNBCs.

Medicinal plants are precious source of bioactive compounds which possess a range of beneficial properties and serve as the major source of medicine for a large proportion of population across the world. Since ancient times, Crataeva nurvala has been used as a vital herb in Ayurvedic system of medicine. In Unani system of medicine the bark of C. nurvala is used as an appetite stimulant and as an agent to decrease the secretion of bile and phlegm. In the present study, the methanol extract of stem bark of C. nurvala was analysed for preliminary phytochemicals and the chemical profiling of the extract was illustrated using gas chromatography mass spectrometry (GC-MS) analysis. The phytochemical analysis revealed that the plant extract contained alkaloids, steroids and triterpenoids. The GC-MS analysis determined the presence of different compounds of biological importance. The identification and characterisation of the phytoconstituents in the extract could pave the way for the discovery of new drugs for various ailments.

Background: The absence of receptors, lack of specific treatment regimen, and emergence of resistance against various currently available anticancer drugs have led to the development of lead molecules from botanicals for the mainstay treatment of triple-negative breast cancer (TNBC). Hordeum vulgare, commonly known as barley, has been reported to have many traditional uses, and the main alkaloid in its germinated seeds, hordenine, has been evaluated for many pharmacological properties. Objectives: The present study aimed to evaluate the anticancer activity of methanol extract of germinated seeds of H. vulgare (MGHV) against MDA-MB-231 TNBC cells, quantify hordenine in MGHV, and derive the probable target of action of the extract and hordenine. Materials and Methods: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, morphological evaluation, acridine orange/ethidium bromide (AO/EB) staining, Hoechst 33258 staining, JC-1 staining, comet assay, and Western blot were conducted to evaluate the anticancer effect. High-performance liquid chromatography (HPLC) analysis was conducted to detect and quantify the presence of hordenine in the extract. Stimulation of extrinsic pathway of apoptosis was evaluated by studying the interaction of hordenine with Caspase-8 using in silico methods. Results: MTT assay revealed significant concentration-dependent cytotoxicity. AO/EB staining exhibited yellow-green fluorescence indicative of early apoptosis. Hoechst 33258 staining showed nuclear marginalization and fragmentation. The results of JC-1 staining showed a combination of red and green fluorescence indicative of partial reduction in mitochondrial membrane potential. Comet assay revealed that the extract did not produce deoxyribo nucleic acid damage. Western blotting analysis did not show any change in the expression of Bcl-2, whereas significant upregulation of Caspase-8 by 2.8 folds was noticed indicative of extrinsic pathway of apoptosis. HPLC analysis detected the presence of hordenine as the major constituent in the extract. Morphological assessment of MDA-MB-231 cells treated with hordenine showed cytotoxic changes similar to that of the extract. In silico studies with hordenine also substantiated the results of Western blot. Conclusion: Thus, it was concluded that MGHV targeted extrinsic pathway of apoptosis due to the presence of hordenine and hence hordenine could evolve as a potent anticancer molecule against TNBC.

Moringa oleifera Lam. is a highly valued medicinal plant in India, especially Kerala. In the present study, antioxidant activity of aqueous extract of leaves of M. oleifera was determined both in-vitro and in-vivo. Male Wistar rats of 3 age groups- 6, 12, and 18 months old were used for in-vivo analysis. In vitro anti-proliferative effect of the extract was carried out in Dalton’s Lymphoma Ascites (DLA) Cells. LCMS-QTOF analysis of the extract was also done to determine the bioactive components present in the extract. Antioxidant activity of M. oleifera leaf showed an IC 50 value of 10.47 ?g/ml and whereas for standard drug, ascorbic acid, it was 19.52 ?g/ml. In-vivo analysis of lipid peroxidation showed a significant reduction of lipid peroxidation in the brains of 12 and 18-months old treated groups. Up to 75% mortality of DLA cancerous cells was observed in-vitro in different concentrations of M. oleifera leaf water extract in a dose-dependent manner, demonstrating its anti-proliferative property. LCMS-QTOF analysis revealed the presence of emodin-8-glucoside in the extract. Molecular docking analysis (Auto Dock Vina) of emodin-8-glucoside with six cancer related proteins showed highest binding affinity with AKT-1 with a binding score of -10.4 kcal/mol, also showed good affinity with NF-kB (p65), Stat-3, Bcl-2, Bcl-xl and c-FLIP. This study helps to choose healthy diet practices to overcome free radical onslaught and cancerous cell proliferation especially in the later stages of life. This can also pave way for the emergence of diet based therapeutic cure for cancer.

Bacterial infections are difficult to treat, especially those infected with resistant bacteria. Due to the haphazard use of antibiotics in humans and animals, methicillin-resistant Staphylococcus aureus (MRSA) has developed resistance to practically all commercially available antibacterial drugs. Exploring natural plant compounds with antibacterial activity could be beneficial in the treatment of infections with MRSA and Staphylococcus aureus. Andrographolide, a labdane diterpenoid derived from Andrographis paniculata, and Hordenine, a phenethylamine alkaloid found in Hordeum vulgare, are reported to have antibacterial activity. In the present study, the antibacterial activity of andrographolide and hordenine was tested against MRSA and S. aureus using the Kirby- Bauer agar disc diffusion method and the modified resazurin microtiter plate assay. The results of the study revealed that both andrographolide and hordenine at various concentrations did not show any zone of inhibition against MRSA and S. aureus. Hordenine showed an MIC of 1000 μg/mL against MRSA and S. aureus, but andrographolide had no effect on both MRSA and S. aureus.

In humans, triple negative breast cancer (TNBC) is a highly truculent form of mammary gland cancer that connexions clinical, pathological and molecular patterns of disease with canine inflammatory mammary carcinoma (IMC). Inflammatory breast cancer with TNBC phenotype has shoddier prognosis and mediocre outcome when juxtaposed with non-inflammatory breast cancer (IBC) with TNBC phenotype. This demands the need for effective therapeutics and targets for its treatment which is currently lacking. Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), a major regulator of inflammation is a potential therapeutic target in TNBC. The present study was instigated to appraise the anticancer properties of methanol extract of Garcinia gummi-gutta (MGG) in MDA-MB-231, a triple negative breast carcinoma cell line. The extract was gaged for its cytotoxic property in tumour necrotic factor- alpha (TNF-α) stimulated MDA-MB-231 cell line, where we obtained a half maximal inhibitory concentration (IC50) value of 50.3 μg/mL. On acridine orange/ ethidium bromide staining, MDA-MB-231 cells treated with the extract unveiled marked morphological and nuclear alterations which were characteristics of apoptosis. In the gene expression study, NF-κB expression was down-regulated in the MGG treated TNF-α stimulated MDA-MB-231 cell line. In conclusion, our present study divulged that the MGG exhibited a concentration dependent cytotoxic activity in TNF-α stimulated MDA-MB-231 breast carcinoma cell lines. Furthermore, our study revealed that the extract was effective in inhibiting the NF-κB signalling pathway which makes it a pledging source for isolating therapeutic molecules for the treatment of inflammatory breast cancer with TNBC phenotype.