IN VITRO MULTIPLE SHOOT REGENERATION STUDIES IN CYPERUS ROTUNDUS L.

Objective: Cyperus rotundus L (Cyperaceae) is one of the highly valuable, potent, multipurpose medicinal herb used in traditional medicine for treatment of several human alignments. To meet the growing herbal needs of pharma industry, we made an attempt on in vitro regeneration of C. rotundus by using rhizome explants.Methods: Rhizome explants were inoculated on both solid and liquid Murashige and Skoog [MS] media supplemented with various concentrations of Kinetin [Kn], 2,4-Dichloro phenoxy acetic acid [2,4-D], Naphthalene acetic acid [NAA], Indole-3-butyric acid [IBA] and activated charcoal for in vitro regeneration of multiple shoots and root induction.Results: Efficient multiple shoots induction (88.9%) was observed on MS liquid media containing 3.0 mg/l 2,4-D+3.0 mg/l NAA+0.5 mg/l Kn after two weeks of explant inoculation with an average number of shoots 9.56±0.35 per explant. Further, we noticed effective root induction (82.3%) from these in vitro grown shoots on root induction media containing 1.0 mg/l IBA with 500 mg/l activated charcoal.Conclusion: Overall, it suggests that this rapid and reliable protocol will be useful for mass multiplication of plantlets and maintenance of germ plasm throughout the year without seasonal constraints. Â

Neoplastic transformation of breast epithelial cells by genotoxic stress

<p>Abstract</p> <p>Background</p> <p>Exposure to genotoxic stresses such as radiation and tobacco smoke can cause increased cancer incidence rate as reflected in an in depth meta-analysis of data for women and breast cancer incidence. Published reports have indicated that exposures to low dose radiation and tobacco smoke are factors that contribute to the development of breast cancer. However, there is a scarcity of information on the combinatorial effects of low dose radiation and tobacco smoke on formation and progression of breast cancer. The combination of these two genotoxic insults can induce significant damage to the genetic material of the cells resulting in neoplastic transformation.</p> <p>Methods</p> <p>To study the effects of low dose ionizing radiation and tobacco smoke on breast cells, MCF 10A cells were treated either with radiation (Rad - 0.1 Gray) or cigarette smoke condensate (Csc - 10 microgram/ml of medium) or a combination of Rad + Csc. Following treatments, cells were analyzed for cell cycle distribution patterns and the ability to extrude the Hoechst 33342 dye. In addition, <it>in vitro </it>invasion and migration as well as mammosphere formation assays were performed. Finally, differential gene expression profiles were generated from the individual and combination treatment.</p> <p>Results</p> <p>Exposure of MCF 10A cells to the combination of radiation plus cigarette smoke condensate generated a neoplastic phenotype. The transformed phenotype promoted increased mammosphere numbers, altered cell cycle phases with a doubling of the population in S phase, and increased invasion and motility. Also, exclusion of Hoechst 33342 dye, a surrogate marker for increased ABC transporters, was observed, which indicates a possible increase in drug resistance. In addition, changes in gene expression include the up regulation of genes encoding proteins involved in metabolic pathways and inflammation.</p> <p>Conclusions</p> <p>The results indicate that when normal breast cells are exposed to low dose radiation in combination with cigarette smoke condensate a phenotype is generated that exhibits traits indicative of neoplastic transformation. More importantly, this is the first study to provide a new insight into a possible etiology for breast cancer formation in individuals exposed to low dose radiation and tobacco smoke.</p

Expression of DDX3 Is Directly Modulated by Hypoxia Inducible Factor-1 Alpha in Breast Epithelial Cells

DEAD box protein, DDX3, is aberrantly expressed in breast cancer cells ranging from weakly invasive to aggressive phenotypes and functions as an important regulator of cancer cell growth and survival. Here, we demonstrate that hypoxia inducible factor-1α is a transcriptional activator of DDX3 in breast cancer cells. Within the promoter region of the human DDX3 gene, we identified three putative hypoxia inducible factor-1 responsive elements. By luciferase reporter assays in combination with mutated hypoxia inducible factor-1 responsive elements, we determined that the hypoxia inducible factor-1 responsive element at position -153 relative to the translation start site is essential for transcriptional activation of DDX3 under hypoxic conditions. We also demonstrated that hypoxia inducible factor-1 binds to the DDX3 promoter and that the binding is specific, as revealed by siRNA against hypoxia inducible factor-1 and chromatin immunoprecipitation assays. Thus, the activation of DDX3 expression during hypoxia is due to the direct binding of hypoxia inducible factor-1 to hypoxia responsive elements in the DDX3 promoter. In addition, we observed a significant overlap in the protein expression pattern of hypoxia inducible factor-1α and DDX3 in MDA-MB-231 xenograft tumors. Taken together, our results demonstrate, for the first time, the role of DDX3 as a hypoxia-inducible gene that exhibits enhanced expression through the interaction of hypoxia inducible factor-1 with hypoxia inducible factor-1 responsive elements in its promoter region

WATER CHLORINATION AND ITS RELEVANCE TO HUMAN HEALTH

Climatic conditions are fundamental to life on earth and their destruction or disturbance by direct or indirect human activities is the greatestthreat to human health. Human life on earth is directly associated with environmental factors such as air†and water.†Pollution of air by toxicsubstances by the activities of mankind has shown to cause serious health issues, including damage to the immune, respiratory, neurological,and reproductive systems, and other health problems like cancer. Water intended for human consumption should be free from microorganismsand toxic substances. The impact and drastic effects of chlorinated water and their impact on human health are poorly studied. Chlorinationis an inexpensive and effective process for disinfecting water worldwide. During the disinfection, the chlorine generates hundreds of differentby-products called chlorination by-products such as trihalomethanes and halo acetic acids (HAA's) at low levels. In this article we address theaction of two HAA's, tri- and di-chloroacetic acid and their impact on the progression of cancer, respiratory disorder, and neurological anomalies. Keywords: Pollution, Chlorination, Chlorine by-products

Multivitamin–Cisplatin Encapsulated Chitosan Nanoparticles Modulate DDX3X Expression in Cancer Cell Lines

Vitamin supplementation during chemotherapy has often been associated with lower recurrence and mortality rates in cancer patients. We had previously demonstrated that the multivitamin (C, D3, and B12)–cisplatin nanoparticle complex—NanoCisVital (NCV)—could alleviate chemotherapy-induced cancer fatigue. Chitosan is frequently used in functional nanomaterials to encapsulate drugs, because it is biodegradable, biocompatible, and non-toxic. The chitosan-based NCVs were prepared, and their physicochemical properties, size, and stability were evaluated before assessing their effect on cancer cell lines. The multivitamin mixture is packed in the core, and cisplatin is loaded at the periphery of the nanoparticle. This encapsulation facilitates the slow and sequential release of peripheral cisplatin and the core multivitamin combination. By increasing the amounts of vitamin and drug-encapsulated nanoparticles in breast and cervical cancer cell lines, the viable cell percentage was calculated. DDX3X promotes cancer cell proliferation, invasion, and metastasis, while Ki-67 promotes active cell proliferation in all cell types. DDX3X is elevated in several cancer types, and breast cancer cells express it abnormally. The Ki-67 protein is a biomarker of cell proliferation that is present throughout all active stages of the cell cycle but undetectable in the resting state. The expression of the DDX3X and Ki-67 genes is altered in NCV-treated cells. This study uses DDX3X and Ki-67 gene expression as a comparative measuring tool for the anti-cancer and cell proliferation effects of cisplatin and vitamins, respectively

Classification and diagnostic prediction of breast cancer metastasis on clinical data using machine learning algorithms

Abstract Metastatic Breast Cancer (MBC) is one of the primary causes of cancer-related deaths in women. Despite several limitations, histopathological information about the malignancy is used for the classification of cancer. The objective of our study is to develop a non-invasive breast cancer classification system for the diagnosis of cancer metastases. The anaconda—Jupyter notebook is used to develop various python programming modules for text mining, data processing, and Machine Learning (ML) methods. Utilizing classification model cross-validation criteria, including accuracy, AUC, and ROC, the prediction performance of the ML models is assessed. Welch Unpaired t-test was used to ascertain the statistical significance of the datasets. Text mining framework from the Electronic Medical Records (EMR) made it easier to separate the blood profile data and identify MBC patients. Monocytes revealed a noticeable mean difference between MBC patients as compared to healthy individuals. The accuracy of ML models was dramatically improved by removing outliers from the blood profile data. A Decision Tree (DT) classifier displayed an accuracy of 83% with an AUC of 0.87. Next, we deployed DT classifiers using Flask to create a web application for robust diagnosis of MBC patients. Taken together, we conclude that ML models based on blood profile data may assist physicians in selecting intensive-care MBC patients to enhance the overall survival outcome

EVALUATION OF IN VITROANTICANCER ACTIVITY AND GC-MS ANALYSIS FROM LEAF SOPHORA INTERRUPTA BEDD

Objective: Sophora interrupta Bedd (Fabaceae) is one of the well-known medicinal herb used in folk medicine for the treatment of cancer and inflammatory associated diseases. In this paper we aimed to evaluate the antioxidant and anti-cancer, properties of aqueous, methanol and n-hexane leaf extracts.Methods: The leaf extracts were analyzed for antioxidant activity using DPPH free radical scavenging assay and anticancer activity by measuring the cell viability using MCF-7 and PC-3 cancer cell lines. GC-MS analysis was performed to identify anti-cancer compounds present in the active leaf extract.Results: The polar solvent extracts showed good antioxidant at 500 μg/ml as related to non-polar solvents. Moreover, methanol extract exhibited highest percentage of cell death, in both MCF-7 (IC50 500 μg/ml) and PC-3 cells (IC50 1000 μg/ml), which is also evident from morphological observations, acridine orange and ethidium bromide dye exclusion assays.Conclusion: Over all, it suggests that leaf methanol extract contain anticancer compounds, which are evident from our GC-MS analysis

Identification of Differentially Expressed Genes in Cervical Cancer Patients by Comparative Transcriptome Analysis

Cervical cancer is one of the most malignant reproductive diseases seen in women worldwide. The identification of dysregulated genes in clinical samples of cervical cancer may pave the way for development of better prognostic markers and therapeutic targets. To identify the dysregulated genes (DEGs), we have retrospectively collected 10 biopsies, seven from cervical cancer patients and three from normal subjects who underwent a hysterectomy. Total RNA isolated from biopsies was subjected to microarray analysis using the human Clariom D Affymetrix platform. Based on the results of principal component analysis (PCA), only eight samples are qualified for further studies; GO and KEGG were used to identify the key genes and were compared with TCGA and GEO datasets. Identified genes were further validated by quantitative real-time PCR and receiver operating characteristic (ROC) curves, and the highest Youden index was calculated in order to evaluate cutoff points (COPs) that allowed distinguishing of tissue samples of cervical squamous carcinoma patients from those of healthy individuals. By comparative microarray analysis, a total of 108 genes common across the six patients’ samples were chosen; among these, 78 genes were upregulated and 26 genes were downregulated. The key genes identified were SPP1, LYN, ARRB2, COL6A3, FOXM1, CCL21, TTK, and MELK. Based on their relative expression, the genes were ordered as follows: TTK > ARRB2 > SPP1 > FOXM1 > LYN > MELK > CCL21 > COL6A3; this generated data is in sync with the TCGA datasets, except for ARRB2. Protein-protein interaction network analysis revealed that TTK and MELK are closely associated with SMC4, AURKA, PLK4, and KIF18A. The candidate genes SPP1, FOXM1, LYN, COL6A3, CCL21, TTK and MELK at mRNA level, emerge as promising candidate markers for cervical cancer prognosis and also emerge as potential therapeutic drug targets

Isolation and characterization of the anticancer compound piceatannol from sophora interrupta bedd

Background: Sophora belongs to the family of Fabaceae and the species in this genus are currently used as a folklore medicine for preventing a variety of ailments including cancer. Our aim was to identify and validate an anticancer compound from Sophora interrupta using multi-spectroscopic, anticancer screening, and molecular docking approach. Methods: The cytotoxicity of the various solvent extracts, petroleum ether, n-butanol, and ethyl acetate (EtOAc) of the S. interrupta root powder was evaluated in a breast cancer cell lines (MCF-7). The extract that had anticancer activity was subjected to column chromatography based on the polarity of the solvents. The anticancer activity of the elution fractions was validated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The isolated metabolite fraction with anticancer activity was run through a C18 column isocratic and gradient high-performance liquid chromatography (HPLC). The structure of the isolated compound was characterized using 1 H nuclear magnetic resonance (NMR), 13 C-NMR, Fourier transform infrared spectroscopy, and liquid chromatography-mass spectrometer methods. Results: The crude EtAOc extract effectively inhibited the proliferation of MCF-7 cells. The column eluted chloroform and EtOAc (4:6) fraction of the EtOAc extract showed significant anticancer activity in the MCF-7 cells compared with normal mesenchymal stem cells. This fraction showed three major peaks in the HPLC chromatogram and the first major peak with a retention time (RT) of 7.153 was purified using preparative-HPLC. The structure of the compound is a piceatannol, which is a metabolic product of resveratrol. Piceatannol formed direct two hydrogen bond interactions between Cys912 (2H), and Glu878 of vascular endothelial growth factor receptor 1 (VEGFR1) with a glide-score (G-score) of −10.193, and two hydrogen bond interactions between Cys919, and Asp1046 of VEGFR2, with a G-score of −8.359. The structure is similar to that of the crystallized protein for VEGFR1 and R2. Conclusions: Piceatannol is a secondary metabolite of S. interrupta that has anticancer activity. Moreover, piceatannol has been isolated for the first time from S. interrupta