Purification and cytotoxicity assay of tomato (Lycopersicon Esculentum) leaves methanol extract as potential anti-cancer agent

This research studied the cytotoxicity effect of tomato leaves methanol extract on cancer cells to address potential therapeutic in MCF-7 breast cancer cell lines and its toxicity towards Vero cells. The extraction was done in a shake flask by 82% methanol, 1:10 (w/v), agitated at 22 °C with 110 rpm within 24 hours. Later, purification process was started by thin layer chromatography (TLC) subjected to determine the best mobile phase for compound separation and collection by means of column chromatography. Next, the effect of purified sample towards MCF-7 breast cancer cell lines and Vero cells were observed using in vitro cytotoxicity assay to indicate its active fractions and its half maximal inhibitory concentration (IC50). Purified sample gave a rational effect towards MCF-7 breast cancer cells with IC50 value of 5.85 μg/ml compared to Taxol with IC50 value of 0.039 μg/ml. The purified sample can also be judged to be harmless as it has IC50 value of 765.6 μg/ml in Vero cells treatment while Taxol gave IC50 value of 0.045 μg/ ml

Purification and cytotoxicity assay of tomato (Lycopersicon Esculentum) leaves methanol extract as potential anti-cancer agent

This research studied the cytotoxicity effect of tomato leaves methanol extract on cancer cells to address potential therapeutic in MCF-7 breast cancer cell lines and its toxicity towards Vero cells. The extraction was done in a shake flask by 82% methanol, 1:10 (w/v), agitated at 22 °C with 110 rpm within 24 hours. Later, purification process was started by thin layer chromatography (TLC) subjected to determine the best mobile phase for compound separation and collection by means of column chromatography. Next, the effect of purified sample towards MCF-7 breast cancer cell lines and Vero cells were observed using in vitro cytotoxicity assay to indicate its active fractions and its half maximal inhibitory concentration (IC50). Purified sample gave a rational effect towards MCF-7 breast cancer cells with IC50 value of 5.85 μg/ml compared to Taxol with IC50 value of 0.039 μg/ml. The purified sample can also be judged to be harmless as it has IC50 value of 765.6 μg/ml in Vero cells treatment while Taxol gave IC50 value of 0.045 μg/ ml

Predicting group of metabolites available in partially purified tomato leaves extract showing anticancer activity by HPLC and FTIR

Previously, tomato leaves have been proved to be one of the potential anticancer agents. High Performance Liquid Chromatography (HPLC) and Fourier transform infrared (FTIR) spectroscopic instrumentation was used to predict the presents of group of metabolites and to assign the possibility of certain absorption bands, in which most of the peaks in partially purified tomato (Solanum lycopersicon) leaves methanol extract are attributable to the specific functional groups. The extraction was carried out in a shake flask by 82% methanol, 1:10(w/v)sample to solvent ratio, agitated at 22°C with 110 rpm within 24 hours. Later, the extract was partially purified by column chromatography. High Performance Liquid Chromatography (HPLC) is used to quantify the number of unknown component presents in the fraction. Then, a FT-IR Bruker Tensor 27 System was used during FTIR data acquisition. The collection of FTIR spectra was carried out at 16 scans with resolution of 4 cm-1using strong apodization in the frequency regions of 4,000–650 cm-1. The results support the premise that HPLC and FTIR spectroscopy are efficient and accurate methods for determining major and minor components presents in the extract

Predicting group of metabolites available in partially purified tomato leaves extract showing anticancer activity by high performance liquid chromatography (HPLC) and Fourier transform infrared (FTIR)

Previously, tomato leaves were proved to be one of the potential anticancer agents. High performanceliquid chromatography (HPLC) and Fourier transform infrared (FTIR) spectroscopic instrumentation were used to predict the presence of group of metabolites and to ascertain the possibility of certain absorption bands, in which most of the peaks in partially purified tomato (Solanum lycopersicon)leaves methanol extract are attributed to the specific functional groups. The extraction was carried outin a shake flask with 82% methanol, 1:10 (w/v) sample to solvent ratio, agitated at 22°C with 110 rpm within 24 h. Later, the extract was partially purified by column chromatography. HPLC was used to quantify the number of unknown component presents in the fraction. Then, a FT-IR Bruker Tensor 27 System was used during FTIR data acquisition. The collection of FTIR spectra was carried out at 16 scans with resolution of 4 cm -1 using strong apodization in the frequency regions of 4,000 to 650 cm-1.The results support the premise that HPLC and FTIR spectroscopy are efficient and accurate methods for determining major and minor components presents in the extract

Microarray and quantitative PCR analysis of gene expression profiles in response to treatment with tomato leaf extract in MCF-7 breast cancer cells

We previously found cytotoxic effects of tomato leaf extract (TLE) on the MCF-7 breast cancer cell line. The aim of this study was to ascertain the molecular mechanisms associated with the usage of TLE as an anticancer agent by microarray analysis using mRNA from MCF-7 breast cancer cells after treatment with TLE for 1 hr and 48 hrs. Approximately 991 genes out of the 30,000 genes in the human genome were significantly (p<0.05) changed after the treatment. Within this gene set, 88 were significantly changed between the TLE treated cells and the untreated MCF-7 cells (control cells) with a cut-off fold change >2.00. In order to focus on genes that were involved in cancer cell growth, only twenty-nine genes were selected, either down-regulated or up-regulated after treatment with TLE. Microarray assay results were confirmed by analyzing 10 of the most up and down regulated genes related to cancer cells progression using real-time PCR. Treatment with TLE induced significant up-regulation in the expression of the CRYAB, PIM1, BTG1, CYR61, HIF1-α and CEBP-β genes after 1 hr and 48 hrs, whereas the TXNIP and THBS1 genes were up-regulated after 1 hr of treatment but down-regulated after 48 hrs. In addition both the HMG1L1 and HIST2H3D genes were down-regulated after 1 hr and 48 hrs of treatment. These results demonstrate the potent activity of TLE as an anticancer agent

Development of process conditions to optimize the extraction of bio-disinfectant from Neem Leaf (Azadirachta Indica)

Central composite design (CCD) under Response surface methodology (RSM) was used in the present study to optimize the extraction parameters for assessing, maximum yield of bio-disinfectant content from Neem leaves extract (NLE). The range of the independent variables, namely solid-solvents ratio (0.05-0.1 w/v), extraction time (1-5 hrs), agitation speed (50-250 rpm) and extraction temperature (35-60 °C) were identified by a first set of four factors and five levels experiments. The optimum conditions for extraction of bio-disinfectant content from NLE were found to be at 75% ethanol with solid solvent ratio 0.067, extraction time 3 hrs, agitation speed 150 rpm and extraction temperature 60 °C. Under these optimized conditions, the maximum activity for bio-disinfectant yield was obtained, which showed 5.57 cm2 inhibition zones for anti-microbial assay. The experimental result was in close agreement with predicted values, thus indicating the suitability of the models developed and the success of RSM in optimizing the extraction conditions

Whole genome resequencing data and grain quality traits of the rice cultivar Mahsuri and its blast disease resistant mutant line, Mahsuri Mutant

In Malaysia, rice mutant varieties that are genetically altered to confer resistance against blast disease have been substantially developed through mutational breeding program. However, due to the limited accessible information on the mutant lines, mutant gene variants corresponding to the disease resistance and other useful agronomic traits are yet to be exploited. Here, we conducted whole genome re-sequencing of blast resistance with kernel elongation traits in mutant line, Mahsuri Mutant (87,639,446 bp raw reads), and its parental line, Mahsuri (85,156,783 bp raw reads) using Illumina Novaseq 6000 sequencing platform with 30x sequencing coverage. The generated genome sequences are aimed to facilitate the discovery of causal mutation and single nucleotide polymorphisms (SNPs) related to the intended traits. The identified SNPs can be further employed to develop allele-specific SNP molecular markers to locate the mutant gene regions. The NGS data obtained (FASTQ format) of the parental and mutant lines have been deposited in the National Center for Biotechnology Information (NCBI) database under sequence read archive (SRA) xwith accession numbers SRR24388814 (Mahsuri) and SRR22952097 (Mahsuri Mutant) respectively