Evaluation of molecular effects associated with apoptosis, tumour progression, angiogenesis and metastasis by a novel combination of drugs with ormeloxifene in triple negative breast cancer cells.

Aim: To investigate the molecular effects of a novel combination [sertraline and plumbagin (comb) with ormeloxifene (Orm)] for anticancer activity in triple negative breast cancer cell line "MDA-MB-231". Methods: The cytotoxic effect of the drugs was analyzed by the MTT assay and nuclear morphological changes by acridine orange/ethidium bromide (AO/EB) staining. Induction of apoptosis by annexin V-FITC staining, active caspase-3 detection and cell cycle analysis were studied in vitro on "MDA-MB-231" cells. The qRT-PCR was done to explore the upregulation and down regulation of targeted genes for angiogenesis, metastasis, tumor suppression and protein folding on the triple negative breast cancer cells. The preliminary anti-angiogenic effect of the drugs was assessed by chorioallantoic membrane (CAM) assay. Results: Orm showed inhibitory effects in "MDA-MB-231" cells in a dose and time dependent manner whereas; the drugs in combination gave better cytotoxic effects in the screening MTT assay. Orm + comb was more effective than Orm alone in eliciting apoptosis as well as inhibited the single cell to grow into a colony. CAM assay using Orm and Orm + comb suggested the anti-angiogenic potential which was further confirmed by the downregulation of VEGF in "MDA-MB-231" cells by qRT-PCR studies. The combination was found to effectively upregulate the expression of P53 and P21 and downregulate the gene expression of zinc finger E-box binding homeobox 1 (ZEB1) and heat shock protein 70 (HSP70) in "MDA-MB-231" cancer cells. Conclusions: Collectively this study reveals the efficacy of Orm + comb as more significant than the clinically used tamoxifen (Tam). The study elucidates the promising novelty of the combination as a potential chemotherapeutic intervention for mitigating the aggressiveness of triple negative breast cancer and it addresses the intrinsic resistance caused by single drug treatments.

Nonhormonal selective estrogen receptor modulator 1-(2-[4-{(3R,4S)-7-Methoxy-2, 2-dimethyl-3-phenyl-chroman-4yl}phenoxy]ethyl)pyrrolidine hydrochloride (ormeloxifene hydrochloride) for the treatment of breast cancer.

Breast cancer is the most common type of diagnosed cancers in women, difficult to treat, and has received international attention because of its aggressive nature and inherent drug resistance mechanisms. Development of a better selective estrogen receptor modulator with good therapeutic profile and less toxicity is very crucial in this scenario. This study was undertaken to evaluate and compare the in vitro and in vivo antitumor activities of ormeloxifene with other clinically used breast cancer drugs. Cytotoxic activity of ormeloxifene was compared with standard drugs, 4-hydroxytamoxifene and Adriamycin. Ormeloxifene (50 µM) concentration showed cytotoxicity of 75% and 82% in MDAMB-231 and 24% and 80% in MCF-7 cells, respectively, after 72 and 144 hr of incubation as displayed by cell viability assay. The same concentration of ormeloxifene was shown to exert 74% caspase-7 activation in MCF-7 cells after 24 hr of incubation by fluorescence resonance energy transfer assay. Cell cycle analysis proved that there was an increase in sub-G1 peak to 64.4% and 33.9% in MDAMB-231 and MCF-7 cells, respectively, after treatment using ormeloxifene (50 µM) for 48 hr. The nonobese diabetic-severe combined immunodeficiency mice bearing tumor xenografts of triple negative MDAMB-231 cells treated with ormeloxifene (3 mg/kg bw) showed significant regression in relative tumor volume compared to control. From the results obtained and as evidenced from prior literature, ormeloxifene in addition to contraceptive use, can be repositioned for the development of an efficacious anticancer drug. These data present the preclinical part of a well concerted effort to place ormeloxifene into further clinical trials.

Thermally conductive thin films derived from defect free graphene-natural rubber latex nanocomposite: Preparation and properties.

Commercially useful rubber products viz. gloves, condoms, tyres, and rubber hoses used in high temperature environments, etc., require efficient thermal conductivity, which increases the lifetime of these products. Graphene can fetch this property, if it is effectively incorporated into the rubber matrix. The great challenge in preparing graphene-rubber nanocomposites is formulating a scalable method to produce defect free graphene and its homogeneous dispersion into polymer matrices through an aqueous medium. Here, we used a simple method to produce defect free few layer (2-5) graphene, which can be easily dispersed into natural rubber (NR) latex without adversely affecting its colloidal stability. The resulting new composite showed large increase in thermal conductivity (480-980%) along with 40% increase in tensile properties and 60% improvement in electrical conductivity. This study provides a novel and generalized approach for the preparation of graphene based thermally conductive rubber nanocomposites.

A clinical study assessing the efficacy of a new variant of the levonorgestrel intrauterine system for abnormal uterine bleeding.

OBJECTIVE: To evaluate the efficacy and safety of a new variant of the levonorgestrel-intrauterine system (LNG-IUS)-Emily-for the treatment of abnormal uterine bleeding (AUB). METHODS: A prospective, multicenter, single-arm, phase 4 study was conducted at six centers in India between July 2012 and August 2013. Eligible women were aged 30-50years, had completed their family, had AUB, and a pictorial bleeding assessment chart (PBAC) score of at least 100. After screening (visit 1) and insertion of the device (visit 2), participants were followed up at 1week, 1month, 3months, and 6months. The primary outcomes were menstrual blood loss (assessed by PBAC) and quality of life (assessed by the EQ-5D-3L questionnaire). RESULTS: Among 63 participants, 45 (71%) completed the study. Mean PBAC score decreased from 238.0±128.7 at screening to 13.1±19.2 at 6months (P<0.001). EQ-5D-3L score increased from 79.0±14.1 at visit 2 to 86.3±9.0 at 6months (P=0.003). No serious adverse events related to the device were reported. CONCLUSION: Among women with AUB, use of the Emily LNG-IUS significantly reduces menstrual bleeding and improves quality of life. Clinical Trials Registry of India: CTRI/2012/07/002843.

Biocompatibility and cytotoxic evaluation of drug-loaded biodegradable guided tissue regeneration membranes.

BACKGROUND: In periodontology, Guided Tissue Regeneration (GTR) is based on the concept of providing a space for entry of cells with regenerative potential into the wound environment to initiate the regeneration of structures lost due to periodontal disease. First generation GTR membranes were primarily non-absorbable membranes like expanded polytetrafluorethylene which required a second surgery for its removal. This led researchers to explore absorbable materials like collagen and synthetic biodegradable polymers to fabricate GTR membranes. In the present study, biodegradable Polylactic acid (PLA) is used to fabricate membranes with the potential to be used for GTR therapy. MATERIALS AND METHODS: Biocompatibility of the PLA membranes were evaluated in a subcutaneous guinea pig model. Antimicrobial effect of the drug-loaded PLA membranes were assessed against a drug-resistant Staphylococcus aureus bacterial isolate. The cytocompatibility of the drug-loaded membranes were evaluated using HeLa cell lines. RESULTS: The PLA membranes were shown to be biocompatible. The drug-loaded PLA membranes showed significant activity against the bacterial isolate. Among the drug-loaded membranes, tetracycline-loaded membrane showed minimal cellular toxicity. CONCLUSION: The results of this study indicate that biodegradable drug-releasing polylactide membranes have the potential to be used for periodontal regeneration. It has the necessary characteristics of a GTR membrane like biocompatibility, space maintaining ability, and tissue integration. Among the various antimicrobial agents loaded in the PLA membranes, tetracycline-loaded membranes exhibited minimal cellular toxicity against HeLa cells; at the same time showing significant activity against a pathogenic bacterium.